Showing papers in "Journal of Reproduction and Development in 2010"

Foundations for smarter cities

Abstract: This paper describes the information technology (IT) foundation and principles for Smarter Cities™. Smarter Cities are urban areas that exploit operational data, such as that arising from traffic congestion, power consumption statistics, and public safety events, to optimize the operation of city services. The foundational concepts are instrumented, interconnected, and intelligent. Instrumented refers to sources of near-real-time real-world data from both physical and virtual sensors. Interconnected means the integration of those data into an enterprise computing platform and the communication of such information among the various city services. Intelligent refers to the inclusion of complex analytics, modeling, optimization, and visualization in the operational business processes to make better operational decisions. This approach enables the adaptation of city services to the behavior of the inhabitants, which permits the optimal use of the available physical infrastructure and resources, for example, in sensing and controlling consumption of energy and water, managing waste processing and transportation systems, and applying optimization to achieve new efficiencies among these resources. Additional roles exist in intelligent interaction between the city and its inhabitants and further contribute to operational efficiency while maintaining or enhancing quality of life.

Monitoring Metabolic Health of Dairy Cattle in the Transition Period

Abstract: This paper reviews the importance of energy metabolism in transition dairy cows, its associations with disease and reproduction, and strategies for monitoring cows under field conditions during this critical time. Essentially all dairy cattle experience a period of insulin resistance, reduced feed intake, negative energy balance, hypocalcemia, reduced immune function, and bacterial contamination of the uterus soon before, or in the weeks after calving. One-third of dairy cows may be affected by some form of metabolic or infectious disease in early lactation. Routine, proactive actions, observations, or analysis are intended to accurately and efficiently provide early detection of problems, to provide an opportunity for investigation and intervention in order to limit the consequences and costs of health problems and reduced animal performance or welfare. Methods of early detection include monitoring of disease and culling records, feed intake, milk production, body condition, and simple metabolic tests. Methods, strategies, and interpretation of measurement of peripartum concentrations of non-esterified fatty acids (NEFA) and beta-hydroxybutyrate (BHB) as indicators of aspects of energy status and disease risk are reviewed. High NEFA (> 0.4 mmol/l) in the last 7 to 10 days before expected calving is associated with increased risk of displaced abomasum (DA), retained placenta, culling before 60 days in milk, and less milk production in the first 4 months of lactation. Subclinical ketosis (serum BHB >1200 to 1400 micromol/l) in the first or second week after calving is associated with increased risk of DA, metritis, clinical ketosis, endometritis, prolonged postpartum anovulation, increased severity of mastitis, and lower milk production in early lactation. There are several validated and practical tools for cow-side measurement of ketosis.

Wireless network cloud: architecture and system requirements

Abstract: With the growth of the mobile communication network, from second-generation to third-generation or fourth-generation networks, technologists in the mobile industry continue to consider advanced wireless network architectures that have the potential to reduce networking costs and provide increased flexibility with respect to network features. In this paper, we propose the wireless network cloud (WNC), a wireless system architecture for a wireless access network. This system makes use of emerging cloud-computing technology and various technologies involved with wireless infrastructure, such as software radio technology and remote radio head technology. Based on open information technology architecture, the WNC provides all the necessary transmission and processing resources for a wireless access network operating in a cloud mode. Note that it is useful to separate the hardware and software for different wireless standards and various services and business models, as well as to meet the new system requirements for emerging wireless technologies, such as collaborative processing at different scales of network use. We analyze several important system challenges involving computational requirements of virtual base stations, I/O throughput, and timing networks for synchronization. Based on current information technologies, we make several suggestions with respect to future system design.

Assessing the Association of the Level of Milk Production with Reproductive Performance in Dairy Cattle

Abstract: There is much debate about possible antagonism between high milk production and reproductive performance. This paper reviews methods of measuring reproductive performance and the association of the level of milk production with pregnancy rate at the herd and individual levels. The main question is whether fertility (the capacity for reproductive function and successful pregnancy) of dairy cows has in fact declined, as opposed to the success of management systems and people at meeting the metabolic, nutritional, housing, and social needs of increasingly productive animals but with no less inherent capacity to achieve and maintain pregnancy; and if fertility really has diminished, the extent to which this decline is caused by increased milk production. There is no doubt that production per cow has increased, but it is unclear how much of this increase can explain the apparent decrease in fertility. It is important to separate the biology of reproductive function from the effects of economically based management decisions about culling and continuation of breeding. Most traditionally-used measures of reproductive performance (calving interval, conception rate, non-return rate) are incomplete or severely biased outcome measures. Both herd and cow-level data should include as much information as possible on confounders of the relationship of production with reproduction. Population or herd-level data should not be used to make inferences about individual-level associations. Considering the quality of data and analytic methods in the published literature, it is not clear if there is any association between higher milk yield and the probability and timing of pregnancy, either among cows at various levels of production in a population at one time, or with increasing production over time.

How to improve the success rate of mouse cloning technology.

Abstract: It has now been 13 years since the first cloned mammal Dolly the sheep was generated from somatic cells using nuclear transfer (SCNT). Since then, this technique has been considered an important tool not only for animal reproduction but also for regenerative medicine. However, the success rate is still very low and the mechanisms involved in genomic reprogramming are not yet clear. Moreover, the NT technique requires donated fresh oocyte, which raises ethical problems for production of human cloned embryo. For this reason, the use of induced pluripotent stem cells for genomic reprogramming and for regenerative medicine is currently a hot topic in this field. However, we believe that the NT approach remains the only valid way for the study of reproduction and basic biology. For example, only the NT approach can reveal dynamic and global modifications in the epigenome without using genetic modification, and it can generate offspring from a single cell or even a frozen dead body. Thanks to much hard work by many groups, cloning success rates are increasing slightly year by year, and NT cloning is now becoming a more applicable method. This review describes how to improve the efficiency of cloning, the establishment of clone-derived embryonic stem cells and further applications.

Introduction to the wire-speed processor and architecture

Abstract: In this paper, we introduce the wire-speed processor (WSP) project, an advanced development project led by IBM Research and the IBM Systems and Technology Group. The WSP represents a generic processor architecture in which processing cores, hardware accelerators, and I/O functions are closely coupled in a system on a chip. The first implementation of the WSP architecture targets applications operating at "wire speed" (i.e., speeds in which the data are transmitted and processed at the maximum speed allowed by the hardware). These applications include those that involve routers, firewalls, intrusion-prevention systems, and other network analytics. The WSP combines 16 multithreaded IBM PowerPC® cores with special-purpose dedicated accelerators optimized for packet processing, security, pattern matching, compression, Extensible Markup Language (XML) parsing, and I/O for networking that provides four 10-Gb/s bidirectional network links. In this paper, we describe the various system components, the underlying design philosophy involving close integration of these components, and the special system features that were developed to achieve this close integration.

Problems with scoring methods and ordinal scales in risk assessment

Abstract: Risk assessment methods based on scoring methods that rate the severity of each risk factor on an ordinal scale are widely used and frequently perceived by users to have value. We argue that this perceived benefit is probably illusory in most cases. We begin by describing a number of common scoring methods currently used to assess risk in a variety of different domains. We then review the literature on the use of ordinal scales in risk analysis, the use of "verbal scales" for eliciting estimates of risks and probabilities, and the extensive research about peculiar human errors when assessing risks. We also supplement this overview with some data of our own. When these diverse kinds of evidence are combined, the case against scoring methods is difficult to deny. In addition to the evidence against the value of scoring methods, there is also a lack of good evidence in their favor. We conclude our overview by reviewing the reasons why risk assessment approaches should describe risk in terms of mathematical probabilities.

Hypoxia is Important for Establishing Vascularization During Corpus Luteum Formation in Cattle

Abstract: Hypoxia-inducible factor 1 (HIF1) has been demonstrated to have critical roles in angiogenesis via transcriptional regulation of angiogenic factors, such as vascular endothelial growth factor (VEGF). In the ovary, angiogenesis is known to occur after ovulation in the developing corpus luteum (CL) in mammals. To determine whether HIF1 participates in angiogenesis in bovine CL, the present study investigated the mRNA and protein expressions of the HIF1 alpha subunit (HIF1A) and VEGF in bovine CL during the estrous cycle. The effects of hypoxia on the expressions of HIF1A protein, VEGF mRNA and VEGF protein in bovine luteal cells were also examined by using a cell culture system. HIF1A mRNA expression was less at the regressed stage than at the other stages, whereas protein expression of HIF1A was highest at the early luteal stage and decreased thereafter. VEGF mRNA expression was highest at the developing luteal stage and decreased thereafter. VEGF protein expression was highest at the early luteal stage and decreased significantly at the regressed luteal stage. Hypoxia increased the amounts of HIF1A protein, VEGF mRNA and VEGF protein in cultured bovine luteal cells. Furthermore, we found that hypoxia inhibited progesterone production in the mid luteal cells, but not in the early luteal cells. The overall findings indicate that HIF1 is one of the factors promoting VEGF-induced angiogenesis during luteal development, and suggest that the hypoxic conditions formed after follicle rupture contribute to establishing luteal vascularization in cattle.

Recent advances in the synchronization of estrus and ovulation in dairy cows.

Abstract: Synchronization programs have become standard components in the current breeding management of cows in the dairy herds of most dairy industries. Many are based on protocols that allow timed inseminations (TAI) so as to circumvent the practical difficulties associated with estrus detection. These difficulties are exacerbated in modern herds of high producing cows either because of increasing herd size in which individual animal monitoring is difficult and often subjective, or because small intensively managed herds are milked in robotic systems that minimize animal: staff interactions. Additional reasons arise from high producing cows having less obvious symptoms of estrus, partly because of housing systems combined with intensive feeding and milking, partly because of higher metabolic clearance rates of reproductive hormones like estradiol and partly because of the increasing prevalence of prolonged post-partum anestrus and reproductive tract pathology. The most recently developed programs include protocols for resynchronization following first or subsequent inseminations. These re-synchronization protocols may involve selected forms of hormonal intervention during the diestrous and pro-estrous periods following TAI, or following pregnancy diagnosis by ultrasound from 28 days after TAI. The latter form of re-synchronization has become increasingly important with the recognition that late embryonic/early foetal death has become a major factor compromising the reproductive performance of high producing Holstein cows in many dairy industries. Although cows detected in estrus without any hormonal treatment before insemination have higher conception rates than those inseminated following synchronization and TAI, the low detection rates combined with embryonic death means that intervals from calving to conception (days open) are usually less when synchronization programs have been successfully implemented. One of the significant factors affecting a program's success is the compliance rate that may sometimes be less than 70%. Almost all programs involve strategically timed injections of prostaglandin F2alpha (PGF) and gonadotropin releasing hormone (GnRH). Injections of an estradiol ester and progesterone supplementation per vaginum may be included in some programs. The basic program is the "Ovsynch" regimen. Numerous variations have been tested and developed. Many involve increasingly complex protocols that increase the risk of non-compliance, none has consistently achieved conception rates that exceed 40% and few have reduced the incidence of embryonic death. These synchronization programs are the best that are currently available. They have not been able to overcome the consequences of lowered fertility associated with high levels of milk yield, forms of nutrition and environmental factors like heat stress that have profound effects on the physiology and metabolism of the high producing dairy cow.

Management of heat stress to improve fertility in dairy cows in Israel.

Abstract: Israel has about 100,000 dairy cows mostly all of Israeli-Holstein-breed, kept in close to 1000 dairy farms. Most farms are distributed along the Mediterranean Sea coast and in the hot internal valleys. According to the Israeli Herd book the average annual milk production, per cow in 2008 was 11,460 kg, with 3.7% fat and 3.2% protein. Israel's climate is considered "subtropical dry" or Mediterranean, characterized by warm and dry summer with day temperatures above 30 C and relative humidity ranging from 50 to 90%. Climatic limitations brought dairy farmers to develop and implement new technologies and management practices that would enable high milk production and reproduction in summers. In the last three decades the Ministry of Agriculture research units, the extension service and dairy farmers conducted a series of trials and surveys in order to develop an efficient cooling system that will obtain and maintain high milk yield and good reproduction during the hot and humid summer. The cooling system commonly used in Israel is based on a combination of frequent direct watering of the cows, followed by forced ventilation air blowing onto the cows. The system was developed in Israel nearly 30 years ago. A typical cycle is five minutes long and consists of 30 sec of watering followed by 4.5 min of forced ventilation. Providing the cows with 5-7 cooling sessions per day, 30-45 min each, allowed cows, producing 25-30 kg of milk per day to maintain their body temperature below 39.0 C, throughout the day time, on a typical Israeli summer day. At the same time, non-cooled cows had high body temperatures (above 39.5 C), during some part of the daytime and returned to normal body temperatures (below 39.0 C), only for a few hours late at night. In an experiment conducted in 1985-86, conception rate (CR) of cows, cooled as described above, was significantly higher than of non-cooled cows (59 vs. 17% and 57 vs. 17%), for first insemination and for all inseminations, respectively. Pregnancy rate (the amount of pregnant cows out of the eligible cows in the herd) calculated for 90, 120 and 150 days after calving differed significantly between the groups, (44, 59 and 73% vs., 5, 11 and 11%), in cooled and non-cooled cows, respectively. CR and pregnancy rates obtained in intensively cooled herds in this experiment were similar to those obtained during the winter of that year, in commercial dairy farms in Israel. Differently from the results described above, when cows in summer were intensively cooled, only for a period of 2 days before and 8 days after A.I, CR failed to improve (31 and 36%), in cooled and non-cooled cows, respectively. These results offer a conclusion that cows must be intensively cooled and must maintain normal body temperatures during the entire day and during the whole summer. i.e. the entire reproductive process from follicular development until implantation of the embryo in the uterus, in order to express cow's full reproductive potential in Israeli summer conditions. The effect of cooling intensity on cow's productive and reproductive traits was studied in a wide survey, during four consecutive years (1998-2001), on 14 farms, averaging 300 milking cows each, all located in the coastal plain of Israel. Farms were categorized into three different groups according to the intensity of summer cooling. "Intensive" (7.5 cumulative cooling hours per day), "Moderate" (4.5 cumulative hours per day) and "No-cooling" at all. CR was 56, 53 and 54%, and 40, 34 and 15%, for primiparous (P < 0.01) and 47, 46 and 43%, and 34, 34 and 17% for multiparous cows (P < 0.01), in the "intensive", "moderate", and "no cooling" groups, in winter and summer, respectively. In another survey based on the Israeli Herd Book data from 2005, using elite yielding herds (with average annual milk production per cow of more than 13,000 kg), the average CR of intensive cooled herds was 39 and 19%, in winter and summer respectively, compared to 39 and 12%, respectively, in non-cooled high yielding herds (P < 0.01). This indicates that intensive cooling in summer can reduce by half the summer drop in CR, even in very high yielding herds. The Ministry of Agriculture extension service, in cooperation with the Israel Cattle Breeders Association (ICBA), developed a computerized report called "Summer to Winter (S:W) Performance Ratio", based on the "Israeli Herd book" data from more than 300 herds. The higher the ratio is for productive and reproductive traits, the better a farm handles summer negative effects on cow's performance. Based on the S:W ratio of each herd in 2007, we quantified the overall effect of intensive cooling in summer on the cow's whole year performance. Data from 24 farms with the highest S:W ratio were compared with data from 24 farms with the lowest S:W ratio. The comparison showed that well cooled cows in Israeli summer added approximately 700 kg of milk to cow's lactation, an increase of 6.5% in its annual production. Summer CR were significantly higher in the highest S:W ratio farms, compared with the lowest ones (27 vs. 19%), and compared to those obtained in same groups in winter (40 vs. 36%), respectively. High S: W ratio herds reached in summer conception rate of 70% of their winter CR, compared to only 50% in the lowest S:W ratio farms inseminated in same period. Trials conducted in the last 10 years show clearly that intensive cooling of high yielding cows (above 45 kg daily) in summer cannot completely eliminate summer decline in CR (as was achieved two decades ago when daily production was less than 30 kg). These high yielding cows despite being intensively cooled could not maintain normal body temperature all day long. This fact brought Israeli researchers to look for hormonal treatments to improve cow's summer fertility, among them elevating post insemination blood progesterone, GnRH treatment at time of insemination to optimise insemination time, improvement of egg quality by elimination of aged follicles produced during heat stress and the use of timed AI and embryo transfer. Part of these treatments improved summer CR when combined with intensive cooling. Cooling Intensification combined with hormonal therapy, management and nutritional practices are expected to minimize the gap between summer and winter CR obtained in Israel in the future.

Relationship between the length of cell cycles, cleavage pattern and developmental competence in bovine embryos generated by in vitro fertilization or parthenogenesis.

Abstract: This study was conducted to study the kinetics of initial cell divisions in relation with the cleavage patterns in viable (with the ability to develop to the blastocyst stage) and non-viable bovine embryos and parthenotes. The kinetics of in vitro development and cleavage patterns were observed by time lapse cinematography. The length of the first and second but not third cell cycle differed significantly between the viable and non-viable embryos after IVF or parthenogenesis. Viable embryos had significantly shorter first and second cell cycles than non-viable ones. The presence of fragments, protrusions and unequally-sized blastomeres was associated with an extended one-cell stage and reduced ability to develop to the blastocyst stage; however, the lengths of the second and third cell cycles were not altered. Oocytes showing direct division from one cell to 3 or 4 blastomeres showed similar developmental ability and embryonic cell numbers to those showing normal division, although, with a high frequency of chromosomal abnormalities. Our results suggest that the differences in the first cell cycles between viable and non-viable embryos were not sperm-related, whereas direct cleavage of 1-cell embryos to 3 or more blastomeres and protrusion formation are related to sperm-driven factors. The length of the first and second cell cycles and the cleavage pattern should be examined simultaneously to predict developmental competence of embryos at early cleavage stages.

Synergistic effects of glutathione and β-mercaptoethanol treatment during in vitro maturation of porcine oocytes on early embryonic development in a culture system supplemented with L-cysteine.

Abstract: Various methods have been used to remove reactive oxygen species (ROS) generated from in vitro culture (IVC) conditions that can cause cell injury or death, including the application of low oxygen (O(2)) tension and the addition of antioxidants. The beneficial effects of antioxidants and O(2) tension on IVC of porcine embryos, however, are controversial among researchers. In this study, we sought to determine the effects and optimal concentrations of antioxidants for the development of porcine embryos in an IVC system. Specifically, we examined the synergistic effects of antioxidants on development to the blastocyst stage in a culture system supplemented with L-cysteine during IVM. Of the antioxidants tested (melatonin, glutathione (GSH), β-mercaptoethanol (β-ME), N-acetylcysteine (NAC) and dithiothreitol (DTT)), addition of GSH (1 mM) or β-ME (25 µM) significantly increased development to the blastocyst stage compared with the controls without antioxidant treatment (22.2 ± 4.2% for 1 mM GSH, 25.9 ± 2.2% for 25 µM β-ME and 12-13% for the control, P<0.05). In addition, the mean cell number per blastocyst was increased by approximately 1.7-fold in the presence of GSH or β -ME. These GSH- and β-ME-induced increases in development to the blastocyst stage and total cell number, however, were not mimicked by melatonin, NAC or DTT, all of which are ROS scavengers. The combination of GSH or β-ME with L-cysteine significantly reduced high O(2) tension-induced ROS production (P<0.05). These results suggest that a combination of 1 mM GSH or 25 µM β-ME with 1 mM L-cysteine could be used for production of high quality porcine blastocysts in IVC systems.

Effect of resveratrol on the development of porcine embryos produced in vitro.

Abstract: The effects of resveratrol (a phytoalexin with a wide variety of pharmacological activities) on pig embryos produced by parthenogenesis and/or in vitro fertilization have been investigated. First, parthenogenetic embryos were generated and cultured in PZM-3 medium supplemented with various amounts of resveratrol (0, 0.05, 0.1, 0.5, 1.0 and 25 microM final concentrations). In the presence of 0.5 microM resveratrol a significantly higher percentage of parthenogenetic embryos reached the blastocyst stage by day 7 compared to non-treated control (43.5+/-6.3% vs. 33.0+/-5.4%; P<0.05). The total cell number of blastocysts also increased as a result of incubation with 0.5 microM resveratrol; the difference was statistically significant between treated and non-treated embryos on day 5 of culture (35.8+/-0.9 vs. 32.1+/-1.1; P<0.05). Resveratrol incubation affected the expression levels of apoptosis-related genes in parthenogenetic blastocysts: the level of Bax transcripts was similar but lower expression of Bcl-2 and Caspase-3 was observed in embryos treated with 0.5 microM resveratrol when compared to control blastocysts (P<0.05). The results of the TUNEL assay were similar in blastocysts developing with or without resveratrol supplementation. In addition, when embryos produced by in vitro fertilization were incubated with 0.5 microM resveratrol, the treatment led to higher frequencies of blastocyst formation (8.6% vs. 13.3%) and elevated total cell numbers (37.1+/-2.4 vs. 43.2+/-1.7) by the end of the 7-day culture period (P<0.05). The results indicate that 0.5 microM resveratrol during culture has a positive effect on early embryonic development of porcine embryos.

A comparison of cryotop and solid surface vitrification methods for the cryopreservation of in vitro matured bovine oocytes.

Abstract: The aim of the present study was to compare the efficacies of the cooling systems of the solid surface (SSV) and Cryotop vitrification methods for cryopreservation of bovine oocytes at the metaphase II stage. The effects of vitrification on oocyte viability, in vitro fertilization (IVF), pronucleus formation and subsequent in vitro development were assessed. In vitro matured (IVM) bovine oocytes were subjected to equilibration and vitrification solutions according to the SSV method, and then the oocytes were vitrified either by dropping onto a cold dry metal surface (SSV group) or by plunging into liquid nitrogen on Cryotop sheets (Cryotop group). Warming was conducted according to the SSV method. Some oocytes were subjected to the cryoprotectants and warming regimen without cooling (Solution control group). The live/dead status of oocytes was evaluated by fluorescein diacetate staining. Live oocytes were subjected to IVF, and the resultant embryos were cultured in vitro. The rates of live oocytes were similar among the Fresh control, Solution control, SSV and Cryotop groups. There was no difference in the rates of fertilization, pronuclear formation and monospermy among these groups. The cleavage rates in the SSV and Cryotop groups (41.6 and 53.2%, respectively) were significantly lower than those in the Fresh control and Solution control groups (65.9 and 61.3%, respectively). The blastocyst rates in SSV and Cryotop groups did not differ (10.3 and 12.8%, respectively); however, they were significantly lower than those in the Fresh control and Solution control groups (36.4 and 24.8%, respectively). The inner cell mass, trophectoderm and total cell numbers in blastocysts did not differ significantly among the Fresh control, Solution control, SSV and Cryotop groups. Our results indicate that IVM bovine oocytes could be cryopreserved successfully using the cooling systems of the Cryotop and Solid Surface Vitrification methods with similar efficacy.

Induction of oocyte maturation by hyaluronan-CD44 interaction in pigs.

Abstract: In most mammals, the oocyte is surrounded with compact multilayers of cumulus cells; these form cumulus-oocyte complexes (COCs). During oocyte maturation, the COCs dramatically expand and this is termed "cumulus expansion". We have previously demonstrated that cumulus expansion is the result of hyaluronan synthesis and accumulation in the extracellular space between cumulus cells in the COCs and that hyaluronan accumulation within the COCs affects oocyte maturation. We have also demonstrated that CD44, the principal hyaluronan receptor, is expressed in the COCs during cumulus expansion and that the interaction between hyaluronan and CD44 appears to be closely related to gap junctional communication of the COCs during the process of meiotic resumption. Based on our previous studies, we review herein that the physiological significance and the molecular mechanism of cumulus expansion for porcine oocyte maturation.

Female fertility in French dairy breeds: current situation and strategies for improvement.

Abstract: In most countries where genetic selection for milk production has been conducted, a continuous degradation of reproductive performances has been observed in dairy cattle. This paper describes the phenotypic evolution of fertility traits in France for the main 3 dairy breeds and the different actions undertaken to counteract this unfavourable evolution of reproductive performance. In 2001, functional traits (including conception rate as a fertility trait) have been included into the French total merit index ISU, to take these traits into account in the selection objective. In addition, a marker-assisted selection (MAS) program has been developed since 2001. Through this approach, a set of QTL related to fertility traits were identified using the phenotypic information for fertility collected on a national basis. Fine mapping of these QTL let to their introduction in 2008 in genomic evaluation and the routine selection for fertility traits in breeding schemes. This approach will evolve in the near future through the inclusion of results of several genomic research programs that were conducted either in station or under field conditions aiming at identifying genes and pathways controlling cattle fertility (based on genomics, transcriptomics and proteomics) and at enhancing phenotyping for reproductive performance. The paper describes the general approach behind these evolutions, the expected benefits of genomic selection and first results. Information on the potential use of reproductive technologies that may improve the efficiency of multiple trait genomic selection are also presented.

Enhancement of Cytoplasmic Maturation of In Vitro-Matured Pig Oocytes by Mechanical Vibration

Abstract: The effects of mechanical vibration during in vitro maturation and/or in vitro culture after artificial activation of pig oocytes on maturation and development were examined. In addition, the optimal conditions were applied to in vitro production of blastocysts derived from miniature pig somatic cell nuclear transfer (SCNT) embryos. Mechanical vibration during in vitro maturation did not affect the rates (60.5 +/- 1.9-69.5 +/- 2.2%) of oocytes reaching the metaphase-II stage. However, the blastocyst formation rates after activation of oocytes matured with mechanical vibration for 5 sec at intervals of 30-60 min or for 10 sec at intervals of 60 min were significantly (P<0.05) higher than those of oocytes matured without mechanical vibration (25.7 +/- 2.0-28.1 +/- 2.7% vs. 12.3 +/- 1.4% and 25.8 +/- 1.8% vs. 15.7 +/- 1.9%, respectively). In contrast, mechanical vibration during in vitro culture after activation did not affect the blastocyst formation (11.6 +/- 5.2-16.5 +/- 3.0%) of oocytes. Mechanical vibration for 5 sec at intervals of 60 min during in vitro maturation of oocytes did not affect fusion (66.8 +/- 3.5-72.1 +/- 3.1%) with miniature pig somatic cells after enucleation. However, the blastocyst formation rate of SCNT embryos was improved (P<0.05) by mechanically vibrating recipient oocytes for 5 sec at intervals of 60 min during in vitro maturation, regardless of the presence or absence of the same treatment during in vitro culture (17.6 +/- 2.5% vs. 9.4 +/- 0.9% and 13.0 +/- 0.3% vs. 7.4 +/- 0.9%, respectively). The results indicated that mechanical vibration enhances the cytoplasmic maturation of in vitro-matured pig oocytes, resulting in improvement of their parthenogenetic development. In addition, it was shown that in vitro maturation of oocytes with mechanical vibration can be applied to efficient production of blastocysts derived from miniature pig SCNT embryos.

Management of Dry and Transition Cows to Improve Energy Balance and Reproduction

Abstract: Fertility of dairy cows has decreased for the past several decades. Measures of energy balance (EB; e.g. change in EB, EB nadir, cumulative negative EB, or duration of negative EB) are related to reproductive performance. Our research group has concluded that modification of diets fed during the dry or transition period are unlikely to have significant effects on postpartum EB and fertility. Rather, more radical alterations in dairy management are needed if energy status of postpartum cows is to be improved. We have examined the potential to alter EB by shortening or eliminating the dry period. In an initial study, the effects of a 56, 28, and 0 d dry period on ovarian dynamics and reproductive performance of dairy cows were examined. Postpartum EB was improved by reducing the dry period; however, only significantly for the 0 d dry period. Cows on the 0 d dry period did not experience negative EB. Improvements in EB were a reflection of lower milk production and greater feed intake. Consistent with the improvements in EB, time to first ovulation and first service, first service conception rate, services per conception, and days open were all improved by reduction of dry period length. Because small animal numbers were used in the study, a second study was conducted on a commercial dairy with many more animals to determine if results from the initial trial could be duplicated. A comparison was made between a 55 and 34 d dry period. Shortening the dry period resulted in fewer days to first ovulation, fewer days open, and a greater percentage of cows pregnant at 150 days in milk, although the improvements in the latter two variables were only significant for cows in their third or greater lactation. A current study is being performed in a large commercial dairy to examine the effects of eliminating the dry period on reproduction utilizing a large commercial dairy.

Effect of NGF on the Motility and Acrosome Reaction of Golden Hamster Spermatozoa In Vitro

Abstract: Motility and fertilizing ability are known to be two important physiological attributes of a mature sperm, yet the mechanism by which spermatozoa mature and become motile remains largely unknown. It has been shown that nerve growth factor (NGF) is a protein essential for the development, maintenance and survival of the peripheral and central nervous systems. However, the presence of high levels of NGF protein and mRNA do not correlate with the innervations by NGF sensitive fibers in tissues such as the testis, prostate and seminal vesicles. These observations have shifted the attention of research to the role of NGF outside of the nervous system. Here, we demonstrate that NGF and its receptors TrkA and p75 are widely expressed in the testis, accessory reproductive organ, and the epididymal sperms. We also show that NGF stimulates two important aspects of sperm functions, motility and the acrosome reaction, in a time- and dose-dependent manner. NGF activated the sperm cell acrosome reaction, while addition of inhibitors specific for MAPK kinase significantly blocked the sperm acrosome reaction. Taken together, our findings suggest that NGF plays an integral role in sperm motility and the acrosome reaction through, at least in part, the MAPK signalling pathway.

Developmental ability of porcine in vitro matured oocytes at the meiosis II stage after vitrification.

Abstract: The aim of the present study was to investigate whether a combination of cytoplasmic lipid removal (delipation) and treatment by a microtubule stabilizer, paclitaxel, would lead to efficient cryopreservation of porcine in vitro matured (IVM) oocytes at the meiosis II (MII) stage. Vitrification and subsequent re-warming and culture of 109 untreated oocytes produced only 9 blastocysts (8.3%). On the other hand, the post-vitrification blastocyst rate was significantly improved (21/113, 18.6%, P<0.05) when oocytes were treated with 1 microM paclitaxel. Oocyte delipation also significantly increased the post-vitrification blastocyst rate compared with the untreated group (15/37, 40.5%, P<0.05). The delipation-and-paclitaxel group exhibited a significantly higher blastocyst rate (34/75, 45.3%, P<0.05) than the paclitaxel group, although it was not significantly higher than that for the delipation group. In transfer experiment, a total of 109 (18.6%) parthenogenetic blastocysts were obtained from 586 oocytes vitrified with the delipation-and-paclitaxel treatment. Transfer of 72 blastocysts to two recipients resulted in 14 (19.4%) somite stage fetuses. In conclusion, we demonstrated for the first time that by removing cytoplasmic lipid droplets from oocytes and performing a microtubule stabilization procedure, vitrified porcine IVM MII-stage oocytes could efficiently develop to the blastocyst stage while retaining the ability to develop into fetuses.

A statistical model for risk management of electric outage forecasts

Abstract: Risk management of power outages caused by severe weather events, such as hurricanes, tornadoes, and thunderstorms, plays an important role in electric utility distribution operations. Damage prediction based on weather forecasts on an appropriate spatial scale can improve the efficiency of risk management by reducing the economic and societal costs associated with restoration efforts. We have developed a method of predicting the number of outages in a fashion that is suitable for use by electric utilities by using a Poisson regression model for spatial data in a Bayesian hierarchical framework. Particular attention is given to building models that incorporate uncertainty in the outage data from the perspective of multiple spatial resolutions and spatial correlation in the outage data. The outage-prediction model was developed using historical outage data from an electric utility company in the northeastern part of the United States. The model is being used by that company in the operations of its overhead electrical distribution system and emergency management operations. We discuss results to date and how the model is being applied. In addition to the damage forecasts, we have developed tools for risk visualization by displaying the uncertainty of the damage forecasts on geographic maps.

The nucleolus in the mouse oocyte is required for the early step of both female and male pronucleus organization.

Abstract: During oocyte growth in the ovary, the nucleolus is mainly responsible for ribosome biogenesis. However, in the fully-grown oocyte, all transcription ceases, including ribosomal RNA synthesis, and the nucleolus adopts a specific monotonous fibrillar morphology without chromatin. The function of this inactive nucleolus in oocytes and embryos is still unknown. We previously reported that the embryo lacking an inactive nucleolus failed to develop past the first few cleavages, indicating the requirement of a nucleolus for preimplantation development. Here, we reinjected the nucleolus into oocytes and zygotes without nucleoli at various time points to examine the timing of the nucleolus requirement during meiosis and early embryonic development. When we put the nucleolus back into oocytes lacking a nucleolus at the germinal vesicle (GV) stage and at second metaphase (MII), these oocytes were fertilized, formed pronuclei with nucleoli and developed to full term. When the nucleolus was reinjected at the pronucleus (PN) stage, most of the reconstructed zygotes cleaved and formed nuclei with nucleoli at the 2-cell stage, but the rate of blastocyst formation and the numbers of surviving pups were profoundly reduced. Moreover, the zygotes without nucleoli showed a disorder of higher chromatin organization not only in the female pronucleus but also, interestingly, in the male pronucleus. Thus, the critical time point when the nucleolus is required for progression of early embryonic development appears to be at the point of the early step of pronucleus organization.

Global Gene Silencing is Caused by the Dissociation of RNA Polymerase II from DNA in Mouse Oocytes

Abstract: As mouse oocytes approach maturity, a global repression of gene transcription occurs. Here, we investigated the involvement of RPB1, the largest subunit of RNA polymerase II (RNAP II), in the regulation of this transcriptional silencing mechanism. Using BrUTP to follow transcription in an in vitro run-on assay, we observed an abrupt decrease in transcriptional activity when oocytes reached their full size (approximately 80 µm). Immunoblotting using antibodies specific for the phosphorylated and unphosphorylated forms of RPB1 revealed that RPB1 is phosphorylated at Ser-2 and Ser-5 in the small growing oocytes in which active transcription occurs. By contrast, in transcriptionally inactive, full-grown oocytes, RPB1 is predominantly unphosphorylated. When we permeabilized the nuclear membrane using Triton X-100 during fixation for immunocytochemistry, the unphosphorylated form of RPB1 diffused out of the nucleus in the full-grown oocytes but still remained there in the small growing oocytes, indicating that RPB1 is not bound to DNA in full-grown oocytes. These results suggest that the immediate cause of global transcriptional silencing is the dissociation of RNAP II from the DNA. We also observed dissociation of RPB1 from the DNA in full-grown oocytes treated with trichostatin A to decondense their chromatin, suggesting that chromatin condensation is not an essential process in gene silencing during oocyte growth.

Formation of Primordial Follicles and Immunolocalization of PTEN, PKB and FOXO3A Proteins in the Ovaries of Fetal and Neonatal Pigs

Abstract: The assembly of primordial follicles and subsequent development and transition of the primordial follicle to the primary follicle are critical processes in ovarian biology. In order to examine follicle formation and development in fetal and neonatal pigs, ovarian samples were obtained from a famous local breed of Chinese pigs, Erhualian pigs, ranging in age from 50 days postcoitum to 1 day postpartum in our current study. Morphological changes in the ovaries of the fetal and neonatal pigs indicated that egg nests were the earliest recognizable gamete cells. The proportion of egg nests decreased from 98.4 to 25.6% and the proportion of single follicles increased from 1.6 to 74.4% between 70 and 90 days postcoitum. The proportions of primordial follicles increased between 70 and 90 days postcoitum but decreased from 90 days postcoitum to 1 day postpartum. Our results suggested that the key stage of primordial follicle formation was between 70 and 90 days postcoitum and that the major stage of transition from primordial follicles into primary follicles was between 90 days postcoitum and 1 day postpartum. Experiments were also conducted to examine the localization of PTEN, PKB and FOXO3A proteins in the porcine ovaries by immunohistochemistry and immunoblotting. The results indicated that PTEN, PKB and FOXO3A were localized in the germ cells of egg nests, cytoplasm of oocytes and granulosa cells of follicles ranging from the primordial to secondary stages and that the staining intensity was weak in granulosa cells but strong in oocytes. The different staining patterns of PTEN, FOXO3A and PKB suggested that these proteins were expressed in a stage- and cell-specific manner during ovarian follicle formation and development in the fetal and neonatal pig.

Framework for building a low-cost, scalable, and secured platform for web-delivered business services

Abstract: Many business services are now being delivered over the Web. This method of delivery is attracting the attention of both small- and medium-sized businesses and providers of application-based services because of its easier access, more affordable fees, and greater efficiency in delivering services to massive numbers of receivers. However, it requires the resolution of several key technical challenges. Data security is of top priority for most users because their data reside on remote servers whose resources are shared among tenants. In addition, such a platform needs to meticulously control the operational and delivery costs so that it remains competitive compared with traditional approaches. We have been developing a low-cost, scalable, and secured platform for Web-delivered business services. The platform enables the delivery of business services over the Web with a set of common business and operational support services, including subscription management, massive multitenancy, federated identity management, Web application security protection, and more. For instance, massive multitenancy deals with the isolation of data and service performance for subscribers. In this paper, we discuss the framework for such a platform, the key challenges for realizing the framework, and the approaches for dealing with the challenges.

Developmental ability of miniature pig embryos cloned with mesenchymal stem cells.

Abstract: The present study compared the developmental ability of miniature pig embryos cloned with fetal fibroblasts (FFs), bone marrow-derived mesenchymal stem cells (MSCs) and differentiated (osteocytes, adipocytes and chondrocytes) MSCs. MSCs were isolated from an approximately 1-month-old female miniature pig (T-type, PWG Micro-pig®, PWG Genetics Korea). MSCs were differentiated into osteocytes, adipocytes and chondrocytes under controlled conditions and characterized by cell surface antigen profile using specific markers. These differentiated or undifferentiated MSCs, as well as FFs of miniature pig, were transferred into enucleated oocytes of domestic pigs. Data from 10 replicates involving 1567 cloned embryos were assessed in terms of developmental rates. The in vitro development rate to the blastocyst stage of embryos cloned with undifferentiated MSCs was significantly (P<0.05) higher than that of embryos cloned with differentiated MSCs and FFs. Surgical transfer of 523 two-cell stage embryos cloned with undifferentiated MSCs into five synchronized domestic pig recipients resulted in 5 cloned miniature pig offspring (1 stillborn and 4 viable) from 2 pregnant recipients. The results imply that MSCs might be multipotent and that they can be used to produce viable cloned miniature pigs that cannot be easily reproduced with differentiated somatic cells.

A behavior-based SMS antispam system

Abstract: Short messaging service (SMS) is one of the fastest-growing telecom value-added services worldwide. However, mobile message spam is a side effect for ordinary mobile phone users that seriously troubles their daily life and, as a result, threatens the revenue of telecom operators. In this paper, we present an SMS antispam system that combines behavior-based social network and temporal (spectral) analysis to detect spammers with both high precision and recall. The system infrastructure and the proposed approximate neighborhood index solution, which solves the scalability issue of social networks, are described in detail. Experimental results demonstrate that our proposed system achieves excellent discrimination between spammers and legitimates, and even with fixed recall at 95%, the online system and offline detection subsystems maintain a precision of about 98% and 99.5%, respectively.

Inhibition of the Ubiquitin-proteasome System Leads to Delay of the Onset of ZGA Gene Expression

Abstract: In mammalian oocytes, the ubiquitin-proteasome system (UPS) is suggested to play important roles in oocyte meiosis resumption, spindle assembly, polar body emission and pronuclear formation by regulating cyclin B1 degradation. However, little is known about the direct relationship between zygotic gene activation (ZGA) and degradation of maternal proteins. Here, we investigated the role of the UPS in the onset of ZGA in early mouse embryos. First, we found degradation of cyclin B1 protein in fertilized oocytes at 1 hpi by western blot analysis and used these oocytes throughout this study. Subsequently, we determined optimal experimental conditions for transient inhibition of proteasomal activity by specific and reversible proteasomal inhibitor MG132 in the G1 phase of the first cell cycle. Under the selected optimal conditions, we subjected transient MG132-treated embryos to reverse transcription (RT)-PCR analysis of expression of four ZGA genes, i.e., the hsp70.1, MuERV-L, eif-1a and zscan4d genes. As a result, we found that onset of expression of the four examined ZGA genes was delayed in both normally developed 2-cell embryos and arrested 1-cell embryos. Our results indicate that proteasomal degradation of proteins by the UPS plays a pivotal role in the molecular mechanisms of ZGA in early mouse embryos.

Seasonal Changes in Spermatogenesis and Immunolocalization of Cytochrome P450 17α-Hydroxylase/c17-20 Lyase and Cytochrome P450 Aromatase in the Wild Male Ground Squirrel (Citellus dauricus Brandt)

Abstract: The purpose of this study was to investigate seasonal changes of spermatogenesis and the cellular localization of P450c17 and P450arom in wild male ground squirrels during the breeding and non-breeding seasons. The testicular weight, testicular size and score count of spermatogenesis from April to September were measured, and histological and immunohistochemical observations of testicular tissues were performed in wild male ground squirrels. In addition, total proteins were extracted from testicular tissue in the breeding and non-breeding seasons and were used for Western blotting analysis for P450c17 and P450arom. There were marked variations in testicular weight, testicular size and score count of spermatogenesis from the breeding season (April) to the non-breeding season (September). Histologically, spermatogonia, primary spermatocytes, secondary spermatocytes and spermatozoa were identified in the breeding season (April). Immunolocalization of P450c17 was detected in Leydig cells and spermatozoa during the breeding season and was only found in Leydig cells during the non-breeding season. The positive signals of P450c17 by Western blotting were both observed in the breeding and non-breeding seasons. Immunolocalization of P450arom was observed in Leydig cells, Sertoli cells and all types of spermatogenic cells including mature-phase spermatozoa in the breeding season, while immunoreactivity for P450arom was not present in the testis of the non-breeding season. With P450arom antibody, a band was also only detected in the breeding season by Western blotting. These results suggest that the seasonal changes in testicular weight and size are correlated with spermatogenesis and immunolocalization of P450c17 and P450arom, and androgen and estrogen may play an important role in the spermatogenesis and testicular recrudescence and regression process.

Temporal and spatial regulation of let-7a in the uterus during embryo implantation in the rat.

Abstract: Mammalian embryonic implantation requires reciprocal interactions between implantation-competent blastocysts and a receptive uterus. Some microRNAs might play a key role during embryo implantation in the mouse, but the let-7a expression profiles in the rat uterus during peri-implantation are unknown. In the study, the expression of let-7a in the uterus during early pregnancy, pseudopregnancy, artificial decidualization and activation of delayed implantation was detected by Northern blotting and in situ hybridization. The effect of steroid hormones on let-7a expression was also detected by Northern blotting and in situ hybridization. Here, we found that the expression level of let-7a was higher on gestation day 6-7 (g.d. 6-7) in rats than on g.d.4-5 and g.d.8-9. Let-7a was specifically localized in glandular and luminal epithelia and decidua. The expression of let-7a was not significantly different in the pseudopregnant uterus and increased significantly in the uteri of rats subjected to artificial decidualization and activation of delayed implantation. Treatment with estradiol-17beta or progesterone significantly increased let-7a expression. Thus, let-7a expression was significantly induced by the process of embryo invasion, and this increased expression level was mainly induced by active blastocysts and decidualization during the window of implantation, implying that let-7a may participate in endometrial decidualization. Steroid hormones, estradiol-17beta or progesterone stimulated let-7a expression.