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Assisted reproduction is one of the significant tools to treat human infertility. Morphological assessment is the primary method to determine sperm and embryo viability during in vitro fertilization cycles. It has the advantage of being a quick, convenient, and inexpensive means of assessment. However, visual observation is of limited predictive value for early embryo morphology. It has led many to search for other imaging tools to assess the reproductive potential of a given embryo. The limitations of visual assessment apply to both humans and animals.One recent innovation in assisted reproduction technology imaging is interferometric phase microscopy, also known as holographic microscopy. Interferometric phase microscopy/quantitative phase imaging is the next likely progression of analytical microscopes for the assisted reproduction laboratory. The interferometric phase microscopy system analyzes waves produced by the light as it passes through the specimen observed. The microscope collects the light waves produced and uses the algorithm to create a hologram of the specimen. Recently, interferometric phase microscopy has been combined with quantitative phase imaging, which joins phase contrast microscopy with holographic microscopy. These microscopes collect light waves produced and use the algorithm to create a hologram of the specimen. Unlike other systems, interferometric phase microscopy can provide a quantitative digital image, and it can make 2D and 3D images of the samples.This review summarizes some newer and more promising quantitative phase imaging microscopy systems for evaluating gametes and embryos. Studies clearly show that quantitative phase imaging is superior to bright field microscopy-based evaluation methods when evaluating sperm and oocytes prior to IVF and embryos prior to transfer. However, further assessment of these systems for efficacy, reproducibility, cost-effectiveness, and embryo/gamete safety must take place before they are widely adopted.Summary SentenceThe ability to quantify images of gamete and embryos rapidly will make it possible to evaluate reproductive cells and their potential fertility.

In beef cattle, 40% of cows that display estrus do not remain pregnant after embryo transfer (ET). Estrus is a physiological behavior influenced by the concentration of sex-steroids. The use of accelerometer devices for estrus detection is a common practice in dairy operations. It remains unclear if this same artificial intelligence software can accurately measure reproductive behavior in beef females. We hypothesized that estrous variables detected by an accelerometer are associated with ovarian, hormonal, pregnancy outcome, and fertility-related variables measured in the cow. Fifty Bos indicus-influenced, cyclic, primiparous cows were fitted with an accelerometer collar (SCR Engineers, Israel) and a breeding indicator (Estrotectâ) to detect estrus after a synchronization protocol in five subsequent experimental replicates (n = 50*5 = 250). Only cows that showed estrus (d 0) remained in the study. The following variables were used from the accelerometer software: Activity Peak, Rumination Nadir, Estrus Duration, and Heat Index, calculated by proprietary algorithms. Dominant follicle (DF) diameter and plasma concentration of estradiol were measured on d -2 (day of progesterone device removal) and on d 0. On d -1 and d 4, endometrial thickness and luminal fluid accumulation were measured by ultrasonography. On d 7, two embryos were transferred/cow. Pregnancies were diagnosed on d 45 and terminated. After 30 d, a new replicate started. Cows included in at least two replicates that remained pregnant 100%, 99-50%, or less than 50% were classified as fertile, sub fertile, and infertile, respectively. Estrus display was recorded for 166 cows (66%). Estrus detection by accelerometer and Estrotect had excellent agreement rate (kappa coefficient 0.76). Cows detected in estrus only by Estrotect had 66% less pregnancy/ET than cows in estrus by both (9/21 vs. 97/145; P = 0.01). The DF diameter on d 0 was not associated with accelerometer variables. Cows with greater and moderate Activity Peak had a greater growth rate of the DF from d -2 to d 0 than lower Activity Peak cows (interaction day*Activity Peak; P = 0.06). Estradiol on d -1 was positively associated with the Activity Peak (P < 0.0001). Cows with lower Activity Peak had a thicker endometrium on both d -1 and d 4 (P=0.02), and greater uterine luminal fluid score on d -1 (P=0.03). Fertility status was not associated with any accelerometer variables. In conclusion, the Activity Peak is associated positively with estradiol concentration and DF growth, however no association was found with pregnancy outcome and fertility. Therefore, the main advantage of accelerometers in beef ranches is the ability to detect estrus at a similar accuracy as breeding indicators, using less labor. Future enhancements in hardware and software to allow prediction of pregnancy outcomes are expected to increase reproductive efficiency in cow-calf operations.

A tissue engineering approach to address craniofacial defects requires a biomaterial that balances macro-scale mechanical stiffness and strength with the micron-scale features that promote cell expansion and tissue biosynthesis. Such criteria are often in opposition, leading to suboptimal mechanical competence or bioactivity. We report the use of a multiscale composite biomaterial that integrates a polycaprolactone (PCL) reinforcement structure with a mineralized collagen-glycosaminoglycan scaffold to circumvent conventional tradeoffs between mechanics and bioactivity. The composite promotes activation of the canonical bone morphogenetic protein 2 (BMP-2) pathway and subsequent mineralization of adipose-derived stem cells in the absence of supplemental BMP-2 or osteogenic media. We subsequently examined new bone infill in the acellular composite, scaffold alone, or PCL support in 10 mm dia. ramus mandibular defects in Yorkshire pigs. We report an analytical approach to quantify radial, angular, and depth bone infill from micro-computed tomography data. The collagen-PCL composite showed improved overall infill, and significantly increased radial and angular bone infill versus the PCL cage alone. Bone infill was further enhanced in the composite for defects that penetrated the medullary cavity, suggesting recruitment of marrow-derived cells. These results indicate a multiscale mineralized collagen-PCL composite offers strategic advantages for regenerative repair of craniofacial bone defects.

AbstractThe aim of this work was to evaluate the effect of bull on the efficacy of different capacitating agents in buffalo. Spermatozoa derived from 4 different bulls were incubated in absence of a capacitating agent, in presence of 0.01 mM heparin and in 20% buffalo estrous serum (BES) for 2 hours Sperm were then exposed to lysophosphatidylcholine, an agent that induces acrosome reaction only on capacitated spermatozoa A double staining technique with Trypan-blue/Giemsa was used to evaluate viability and acrosome status of spermatozoa fixed in 37% formaldehyde. The efficiency of capacitation was evaluated by assessing the percentage of acrosome-reacted (AR) spermatozoa in the treated groups. A bull effect on sperm capacitation in vitrowas demonstrated, as indicated by differences in the percentage of AR sperm among bulls regardless of the treatment. In particular when heparin was used as capacitating agent bull C gave higher percentages of AR sperm than bull A (22.7 vs. 14.0%, respectively) with intermediate values for bulls B and D (16.7 and 19.1%, respectively), whereas when BES was used bull D was the most efficient one (22.1, 21.9, 25.0, and 35.0% for bulls A, B, C and D, respectively).

AbstractThe aim of this work was to evaluate chromatin and spindle organization of buffalo and bovine in vitromatured oocytes after vitrification/warming by Cryotop and after their exposure to cryoprotectants (CP). In vitromatured oocytes were vitrified/warmed and exposed to the vitrification/warming solutions containing ethylene glycol (EG), dimethyl sulfoxide (DMSO) and sucrose as CP. Two hours after warming, oocytes were fixed and immunostained for microtubules and nuclei and examined by fluorescence microscopy. Data were analyzed by Chi Square test. A higher percentage of Telophase II stage oocytes was found in the toxicity (26 and 34% in bovine and buffalo) and the vitrification groups (13 and 7% in bovine and buffalo) compared to the control, indicating occurrence of activation. An increased percentage of oocytes with abnormal spindle and chromosome organization was found in oocytes exposed to CP (24 and 13% in bovine; 32 and 30% in buffalo respectively) and in those vitrified (26 and 31% in bovine; 26 and 29% in buffalo respectively) compared to the control (0 in bovine and 2.5 % in buffalo).