Intensity of estrus following an estradiol-progesterone-based ovulation synchronization protocol influences fertility outcomes

The objective of this study was to examine the association between increased physical activity at the moment of timed artificial insemination (AI), detected by an automated activity monitor (AAM), and fertility outcomes. This paper also investigated factors affecting estrous expression in general. A total of 1,411 AI events from 1,040 lactating Holstein cows were recorded, averaging 1.3 ± 0.6 (±standard deviation) events per cow. Activity (measured as steps/h) was monitored continuously by a leg-mounted AAM located on the rear leg of the cow. Ovulation was synchronized by a timed AI protocol based on estradiol and progesterone. Ovarian ultrasonography was performed in all cows on d ?11 (AI = d 0) and in a subset of cows on d 0 (n = 588) and d 7 (n = 819) to determine the presence of a corpus luteum and follicles. The body condition score (1 to 5 scale) was assessed on d 0 and a blood sample was collected for progesterone measurement on d 7. Using the AAM, an estrus event was determined when the relative increase (RI) in physical activity of the cow exceeded 100% of the baseline activity. The physical activity was classified as strong RI (≥300% RI), moderate RI (100–300% RI), or no estrus (<100% RI). Milk production was measured daily and averaged between d ?11 and 0. Pregnancy was diagnosed at 32 and 60 d post-AI and pregnancy losses were calculated. The mean RI at estrus was 328.3 ± 132.1%. Cows with strong RI had greater pregnancy per AI than those with moderate RI and those that did not express estrus (35.1 vs. 27.3 vs. 6.2%). When including only cows that successfully ovulated after timed AI, those that displayed strong intensity RI still had greater pregnancy per AI than those with moderate intensity RI or those that did not express estrus (45.1 vs. 34.8 vs. 6.2%). Cows expressing strong RI at timed AI had greater ovulation rates compared with moderate RI and cows that did not express estrus (94.9 vs. 88.2 vs. 49.5%). Furthermore, pregnancy losses were reduced in cows with strong RI compared with cows expressing moderate RI (13.9 vs. 21.7%). Cows with a strong RI at estrus were more likely to have a corpus luteum at the beginning of the protocol and had greater concentration of progesterone 7 d post-AI. Multiparous cows expressed lower RI compared with primiparous cows. Cows with lower body condition score tended to have decreased RI at estrus. No correlation between estrous expression and pre-ovulatory follicle diameter was observed. Also, no correlation was observed between milk production at AI and RI. In conclusion, strong intensity RI of estrus events at timed AI was associated with improved ovulation rates and pregnancy per AI, and reduced pregnancy losses. These results provide further evidence that measurements of estrous expression can be used to predict fertility at the time of AI and possibly be used as a tool to assist decision making strategies of reproduction programs.     

Multiphoton Physics

Abstract

We discuss recent developments in the generation of ultra-intense laser pulses and show how these can be used to study matter in the presence of electric fields that compete with that due to the nucleus. This has led to a very rapid growth in a wide range of multiphoton studies, many of which are represented in this volume.     

Comparison of methods for fluorescent detection of viable, dead, and total Escherichia coli O157:H7 cells in suspensions and on apples using confocal scanning laser microscopy following treatment with sanitizers

The influence of treating Escherichia coli O157:H7 cells labeled with an enhanced green fluorescent protein (EGFP) plasmid with 20 microg/ml active chlorine, 100 mg/ml hydrogen peroxide, and 80 mg/ml acetic acid on fluorescence intensity was determined. In addition, fluorescent staining methods to differentiate viable and dead E. coli O157:H7 cells on the cuticle of Red Delicious cv. apples following treatment with water or 200 microg/ml active chlorine were evaluated. Suspensions of E. coli O157:H7 EGFP+ cells were exposed to chemical treatment solutions for 0, 30, 60, 120, or 300 s before populations (log10 cfu/ml) were determined by surface plating, and fluorescence intensities of suspensions and individual cells were measured using spectrofluorometry and confocal scanning laser microscopy (CSLM), respectively. The relative fluorescence intensity of suspensions and individual cells changed upon exposure to various treatments. Results indicate that the use of EGFP to tag E. coli O157:H7 may not be appropriate for investigations seeking to microscopically differentiate viable and dead cells on produce following surface treatment with sanitizers. SYTOX Orange and SYTOX Green nucleic acid stains fluorescently labeled dead E. coli O157:H7 cells attached to apple cuticles more intensely than did propidium iodide. A cross-signal occurred between CSLM photomultipliers when examining tissues treated with SYTOX Orange to detect dead cells and antibody labeled with Alexa Fluor 488 to detect total (dead and viable) cells. Because of the possibility of cross-signal resulting in an overestimation of the number of dead cells on apples and, perhaps, other produce treated with these stains, SYTOX Green is preferred to detect dead cells and antibody labeled with Alexa Fluor 594 is preferred to detect the total number of cells on apple surfaces following treatment with sanitizers. The performance of SYTOX Green in combination with Alexa Fluor 594 to detect dead and total cells of E. coli O157:H7 on other produce remains to be determined.     

Testing homogeneous spreadsheet grids with the "what you see iswhat you test" methodology

Although there has been recent research into ways to design environments that enable end users to create their own programs, little attention has been given to helping these end users systematically test their programs. To help address this need in spreadsheet systems (the most widely used type of end-user programming language), we previously introduced a visual approach to systematically testing individual cells in spreadsheet systems. However, the previous approach did not scale well in the presence of largely homogeneous grids, which introduce problems somewhat analogous to the array-testing problems of imperative programs. We present two approaches to spreadsheet testing that explicitly support such grids. We present the algorithms, time complexities, and performance data comparing the two approaches. This is part of our continuing work to bring to end users at least some of the benefits of formalized notions of testing without requiring knowledge of testing beyond a naive level     

Metal ion and guanine nucleotide modulations of agonist interaction in G-protein-coupled serotonin1A receptors from bovine hippocampus

1. The serotonin type 1A (5-HT1A) receptors are members of a superfamily of seven transmembrane domain receptors that couple to GTP-binding regulatory proteins (G-proteins). We have studied the modulation of agonist binding to 5-HT1A receptors from bovine hippocampus by metal ions and guanine nucleotide. 2. Bovine hippocampal membranes containing the 5-HT1A receptor were isolated. These membranes exhibited high-affinity binding sites for the specific agonist [3H]OH-DPAT. 3. The agonist binding is inhibited by monovalent cations Na+, K+, and Li+ in a concentration-dependent manner. Divalent cations such as Ca2+, Mg2+, and Mn2+, on the other hand, show more complex behavior and induce enhancement of agonist binding up to a certain concentration. The effect of the metal ions on agonist binding is strongly modulated in the presence of GTP-gamma-S, a nonhydrolyzable analogue of GTP, indicating that these receptors are coupled to G-proteins. 4. To gain further insight into the mechanisms of agonist binding to bovine hippocampal 5-HT1A receptors under these conditions, the binding affinities and binding sites have been analyzed by Scatchard analysis of saturation binding data. Our results are relevant to ongoing analyses of the overall regulation of receptor activity for G-protein-coupled seven transmembrane domain receptors.     

The effects of biological and social risk factors on special education placement: birth weight and maternal education as an example

The characteristics of objective lenses and Ca2+-sensitive probes were examined for imaging with a two-photon laser-scanning microscope (TP-LSM). The brightness of the images of beads taken by different objectives greatly varied and depended predominantly on their numerical aperture (NA) and less on transmittance and chirping effects. Lateral and axial resolutions, dx and dz, defined as the half decay length of fluorescence intensity of the image of a spherical bead (0.3 m) were 0.12 and 0.42 microm (objective; 40x/0.75). They are far better than those of confocal microscopes (0.3 and 1.5 microm, respectively) measured similarly (Kuba et al., 1994). dx linearly increased with an increase in 1/NA, while dz linearly increased with an increase in n/(NA)2 (n, refractive index) except for an objective of large NA (1.3). The coverslip compensation of objective lenses greatly affected the shape of the X-Z scanned images of 5.0 microm beads as well as resolutions, indicating a large effect of spherical aberration. Two-photon excitation spectra of Ca2+-sensitive fluorescent probes, indo-1, fura-2 and Oregon Green BAPTA-1, lied in a wavelength range shorter than twice that activated by one-photon absorption, while emission spectra were unchanged. Three-dimensional images of a cultured hippocampal neurone loaded with Oregon Green BAPTA-1 showed fine structures of spines, dendrites and axons, while imaging with FM1-43 localized presynaptic boutons and demonstrated synaptic vesicle turnover. Dyes bleached little during the recording of 100 sectioned images. These characteristics of TP-LSM as well as its ability to image deeper tissues provide excellent means to study dynamic, spatial changes in intracellular substances and structures. To achieve the good performance of a TP-LSM, however, the relevant usage of appropriate objectives and fluorescent probes are required.     

Scaling up visual programming languages

The directness, immediacy, and simplicity of visual programming languages are appealing. The question is, can VPLs be effectively applied to large scale programming problems while retaining these characteristics. In scaling up, the problem is how to expand applicability without sacrificing the goals of better logic expression and understanding. From a size standpoint, scaling up refers to the programmer's ability to apply VPLs in larger programs. Such programs range from those requiring several days' work by a single programmer to programs requiring months of work, large programming teams, and large data structures. From a problem domain standpoint, scaling up refers to suitability for many kinds of problems. These range from visual application domains-such as user interface design or scientific visualization-to general purpose programming in such diverse areas as financial planning, simulations, and real time applications with explicit timing requirements. To illustrate the scaling up problem, we discuss nine major subproblems and describe emerging solutions from existing VPL systems. First, we examine representation issues, including static representation, screen real estate, and documentation. Next, we examine programming language issues-procedural abstraction, interactive visual data abstraction, type checking, persistence, and efficiency. Finally, we look at issues beyond the coding process     

Discrepancies in normative data between Lunar and Hologic DXA systems

Many studies have shown the high correlation between Lunar and Hologic DXA bone mineral density (BMD) measurements despite differences in absolute calibration. However, in clinical practice, raw BMD values (in g/cm2) are not normally used for assessing skeletal status and fracture risk. Instead, the BMD values are expressed in terms of the number of standard deviations above or below the young normal value (commonly referred to as the T-score). If the normative populations of the various systems are consistent, the standard deviation scores should also be consistent. For this reason, the World Health Organization (WHO) recently established diagnostic criteria for osteoporosis based on T-scores and not BMD. However, few studies have compared the instruments in terms of their standard deviation scores. In this study, we used linear regression to compare T-scores in 83 women at L1-4 and 120 women at the femoral neck obtained on a Lunar DPX and a Hologic QDR-1000/W system. patient BMD and T-score measurements were highly correlated between the two systems (r > 0.95). No clinically significant difference in L1-4 T-scores was seen (less than 0.1 SD). However, linear regression analysis confirmed a systematic difference of 0.9 SD between the femoral neck T-scores. This discrepancy is caused by: (1) differences in the normal populations, and (2) differences in statistical models used to determine the young normal mean and standard deviation. In an attempt to correct the discrepancy, the female young normal mean and standard deviation were recalculated for the femoral neck using published epidemiological data from NHANES and existing DXA cross-calibration equations. The Hologic young normal value (mean +/- SD) was redefined as 0.85 +/- 0.11 g/cm2, while the Lunar value was redefined as 1.00 +/- 0.11 g/cm2. When the femoral neck T-scores for the study population were recalculated on the basis of these new values, the results were equivalent between manufactures, effectively eliminating the discrepancy. However, the revised values should be confirmed by additional measurements in young normal adults.