Abstract
EPPIN (epididymal protease inhibitor) is a mammalian conserved sperm-binding protein displaying an N-terminal WFDC (whey-acidic protein four-disulfide core) and a C-terminal Kunitz protease ...inhibitor domains. EPPIN plays a key role in regulating sperm motility after ejaculation via interaction with the seminal plasma protein SEMG1 (semenogelin-1). EPPIN ligands targeting the SEMG1 binding site in the Kunitz domain are under development as male contraceptive drugs. Nevertheless, the relative contributions of EPPIN WFDC and Kunitz domains to sperm function remain obscure. Here, we evaluated the effects of antibodies targeting specific epitopes in EPPIN’s WFDC (Q20E antibody, Gln20-Glu39 epitope) and Kunitz (S21C and F21C antibodies, Ser103-Cys123 and Phe90-C110 epitopes, respectively) domains on mouse sperm motility and fertilizing ability. Computer-assisted sperm analysis showed that sperm co-incubation with S21C antibody (but not F21C antibody) lowered progressive and hyperactivated motilities and impaired kinematic parameters describing progressive (straight-line velocity; VSL, average path velocity; VAP and straightness; STR) and vigorous sperm movements (curvilinear velocity; VCL, amplitude of lateral head movement; ALH, and linearity; LIN) compared with control. Conversely, Q20E antibody-induced milder inhibition of progressive motility and kinematic parameters (VAP, VCL and ALH). Sperm co-incubation with S21C or Q20E antibodies affected in vitro fertilization as revealed by reduced cleavage rates, albeit without changes in capacitation-induced tyrosine phosphorylation. In conclusion, we show that targeting specific epitopes in EPPIN Kunitz and WFDC domains inhibits sperm motility and capacitation-associated events, which decrease their fertilizing ability; nevertheless, similar observations in vivo remain to be demonstrated. Simultaneously targeting residues in S21C and Q20E epitopes is a promising approach for the rational design of EPPIN-based ligands with spermostatic activity.
This article reports that europium-doped strontium titanate (SrTiO3:Eu3+) was successfully synthesized using a co-precipitation method at room temperature with processing in a microwave-hydrothermal ...system at 140°C for 30min. Phase composition and structure were examined using X-Ray Diffraction, and Fourier-Transform Raman spectroscopy, revealing a cubic structure with a Pm3m space group. The optical properties were investigated by ultraviolet-visible absorption and photoluminescence, which showed that red emissions originate from Eu3+ transitions. Field emission scanning electron microscopy revealed spherical-like Eu3+ doped SrTiO3 nanoparticles.
Rapid growth in broilers is associated with susceptibility to metabolic disorders such as pulmonary hypertension syndrome (ascites) and sudden death. This study describes a genome search for QTL ...associated with relative weight of cardio respiratory and metabolically important organs (heart, lungs, liver and gizzard), and hematocrit value in a Brazilian broiler-layer cross. QTL with similar or different effects across sexes were investigated. At 42 days of age after fasted for 6 h, the F
2
chickens were weighed and slaughtered. Weights and percentages of the weight relative to BW42 of gizzard, heart, lungs, liver and hematocrit were used in the QTL search. Parental, F
1
and F
2
individuals were genotyped with 128 genetic markers (127 microsatellites and 1 SNP) covering 22 linkage groups. QTL mapping analyses were carried out using mixed models. A total of 11 genome-wide significant QTL and five suggestive linkages were mapped. Thus, genome-wide significant QTL with similar effects across sexes were mapped to GGA2, 4 and 14 for heart weight, and to GGA2, 8 and 12 for gizzard %. Additionally, five genome-wide significant QTL with different effects across sexes were mapped to GGA 8, 19 and 26 for heart weight; GGA26 for heart % and GGA3 for hematocrit value. Five QTL were detected in chromosomal regions where QTL for similar traits were previously mapped in other F
2
chicken populations. Seven novel genome-wide significant QTL are reported here, and 21 positional candidate genes in QTL regions were identified.
Contents
This study aimed to establish a culture system that improves the in vitro development of caprine preantral follicles. In a first experiment, follicles were encapsulated as a single unit per ...bead and cultured singly or in groups or with five follicles in the same alginate (ALG) bead for 18 days. In a subsequent experiment, the “five follicles per bead” design was chosen to culture in ALG, fibrin–alginate (FA) or hyaluronate (HA) for 18 days. In a third experiment, we chose the five follicles per bead in FA to culture for 30 days. The culture set‐up of five follicles per ALG bead increased antrum formation and follicle diameter compared to the other culture designs (p < .05). Moreover, under this condition, 44.44% of the oocytes from in vitro cultured preantral follicles reached meiotic resumption. A significant increase of follicle diameter occurred in attachment system and FA (p < .05), but the ALG condition reached the highest among all groups on day 18 (p < .05). Follicles encapsulated in matrix produced more estradiol and progesterone than attachment system (p < .05). The expression of MMP‐9 mRNA was higher in FA than in other groups (p < .05) and similar to antral follicles from in vivo control (p > .05). Only FA group resulted in oocytes matured. After 30 days, oocytes from preantral follicles in vitro grown in FA developed to eight‐cell parthenotes. In conclusion, a culture system using FA supported the development of caprine preantral follicles cultured in group and included in the same bead of hydrogel, improving the oocyte maturation and producing parthenotes.
The present study evaluated the effects of heat stress on reproductive parameters of hairy rams. Six animals were subjected to scrotal insulation during four consecutive nights (6 PM – 6 AM). Day (D) ...0 was the first day of insulation. Scrotal circumference increased from 30.5 ± 0.3 cm (at pre-insulation) to 31.8 ± 0.4 cm on D4, decreased 3.9 cm on D28, returning to 30.6 ± 0.6 cm on D57. Sperm concentration decreased from 3.7 ± 0.12 ×109 sperm/mL before insulation to 2.6 ± 0.1 ×109 on D23, returning to normal on D57. Sperm motility averaged 75 ± 2.9% before insulation, was undetectable on D23, and became normal on D77. Sperm with normal morphology reached 5.9 ± 2.6% on D35 but recovered (86.8 ± 2.1%) on D91. Sperm DNA integrity decreased from 86.5 ± 4.7% before insulation to 11.1 ± 3.7% on D63, returning to pre-insulation values on D120. Sperm BSP immunostaining was reduced after scrotal insulation. Variations in seminal protein abundances coincided with changes in sperm parameters. Seminal plasma superoxide dismutase, carboxypeptidase Q-precursor and NPC intracellular cholesterol transporter 2 decreased on D18, returning to normal after D28. Albumin, inhibitor of carbonic anhydrase precursor, EGF-like repeat and discoid I-like domain-containing protein 3 and polymeric immunoglobulin receptor increased after insulation. In summary, intermittent scrotal insulation drastically altered ram sperm attributes and seminal proteins, especially those associated with oxidative stress. Knowledge of animal´s response to thermal stress is vital in the scenario of climate changes.
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•Heat stress reduced ram testis size, sperm concentration and motility.•Sperm DNA integrity and BSP binding and seminal proteins were critically altered.•Variations in seminal protein expressions coincided with changes in sperm criteria.•Altered seminal proteins were mainly those associated with oxidative stress.•Know animal response to heat stress is vital in the scenario of climate changes.
Carbynes have gained highlight due to their unique properties and applications in molecular electronics exhibiting resonant tunnel effect. We studied the electronic transport through a molecular ...tunnel junction composed of two polyyne-type carbynes separated by distance (4 Å ≥ d
n
≥ 2.1 Å) higher than bond length (d
n
> d
C-C
) ranging from 0.1 Å based on Density Functional Theory/Non-Equilibrium Green's Function (DFT/NEGF). The frontier molecular orbitals and bond lengths of the polyyne (d
C-C
≈ 1.36 Å, d
C
≡C
≈ 1.26 Å) at 0.0 V become equal to those in cumulene-type carbynes (d
C=
C
≈ 1.28 Å) at 0.9 V occurring a semiconductor-metal transition for d
n
< 4 Å. I-V (dI/dV-V) curve presents switching behaviour up to 0.6 V and after a diode behaviour favoured as d
n
decreases, that is, to 4.0 Å is 3.3 nA (5 nS) while for 2.1 Å is 3750 nA (1500 nS) showing metallic nature. The log(I)-V log(dI/dV)-V curve exhibits the behaviour for low bias voltage while the I-d
n
curve is exhibited for high bias voltage. The exponential expression for G(d) fits perfectly with the G
max
result obtained with DFT/NEGF calculations. Transition Voltage Spectroscopy (TVS) confirms rectangular barrier and direct tunneling for V
d-s
< ϕ
B
/e playing a tunneling rule for electronic transport of these 1-D systems.
We evaluated the effects of maternal dietary flaxseed during lactation on milk composition, body composition and sexual function of the adult female offspring. The dams were fed a control casein diet ...(C) or flaxseed diet (F, 25%) throughout lactation. F mothers showed higher serum 17β-estradiol (E
2) and leptin at weaning. F mother’s milk had lower total cholesterol (TC) and higher E
2 and leptin. The offspring of F dams showed lower body mass (BM), body fat mass (BFM), visceral fat mass (VFM), TC and triglycerides (TG) and higher serum leptin and E
2 at 21
days. F offspring showed delayed puberty onset. At 150
days, these offspring presented higher BFM, VFM, TC, TG, E
2 and lower relative uterine weight and lower progesterone. In conclusion, flaxseed during lactation did affect the lipid profile, adipose tissue and sexual function in adulthood, probably due hyperestrogenism and hyperleptinemia at weaning.
We report on the surface patterning of carbon fiber-reinforced polymer (CFRP) composites using femtosecond laser interferometry. The effect of experimental processing parameters, such as the pulse ...energy and scanning speed, on the quality of the patterns is studied. Using the appropriate processing parameters, a selective removal of the epoxy resin can be achieved, leaving the carbon fibers exposed and textured with the desired pattern. The period of the patterns can be controlled by changing the distance between the two interfering beams. On the other hand, the amplitude of the patterns can be controlled by changing the pulse energy or the number of laser pulses applied. In addition, sub-micron ripples are created on the carbon fibers surface allowing multiscale surface modification which may contribute to improve bonding between CFRP parts.
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