Prediction of 24-h and 6-h Periods before Calving Using a Multimodal Tail-Attached Device Equipped with a Thermistor and 3-Axis Accelerometer through Supervised Machine Learning.

In this study, we developed calving prediction models for 24-h and 6-h periods before calving using data on physiological (tail skin temperature) and behavioral (activity intensity, lying time, posture change, and tail raising) parameters obtained using a multimodal tail-attached device (tail sensor). The efficiencies of the models were validated under tethering (tie-stall) and untethering (free-stall and individual pen) conditions. Data were collected from 33 and 30 pregnant cattle under tethering and untethering conditions, respectively, from approximately 15 days before the expected calving date. Based on pre-calving changes, 40 features (8 physiological and 32 behavioral) were extracted from the sensor data, and one non-sensor-based feature (days to the expected calving date) was added to develop models using a support vector machine. Cross-validation showed that calving within the next 24 h under tethering and untethering conditions was predicted with a sensitivity of 97% and 93% and precision of 80% and 76%, respectively, while calving within the next 6 h was predicted with a sensitivity of 91% and 90% and precision of 88% and 90%, respectively. Calving prediction models based on the tail sensor data with supervised machine learning have the potential to achieve effective calving prediction, irrespective of the cattle housing conditions.

Plasma profile of follicle-stimulating hormone and sex steroid hormones after a single epidural administration of follicle-stimulating hormone via caudal vertebrae in Holstein dry cows.

The conventional follicle-stimulating hormone (FSH) treatment for bovine superstimulation involves multiple intramuscular injections, which is stressful for animals and onerous. We herein investigated whether a single epidural injection of porcine FSH (pFSH) can induce superovulation and peripheral concentrations of pFSH and steroid hormones after the treatment in Holstein dry cows. We intramuscularly administered pFSH twice daily to three cows for 3 days (control) or a single epidural pFSH administration (epidural). Numbers of follicles (≥10 mm in diameter) at estrus and corpora lutea at luteal phase were counted by ultrasonography. Blood was sampled from 0 to 104 h after the first pFSH administration and plasma pFSH, progesterone, androstenedione, testosterone, and estradiol-17ß concentrations were measured. Numbers of follicles (control: 18.3 ± 7.5, epidural: 15.7 ± 4.0; mean ± SD) and corpora lutea (control: 7.3 ± 4.2, epidural: 8.0 ± 2.6) were similar between both treatments. Plasma pFSH concentrations were higher in epidural than in control (p < 0.01). Although no significant differences were observed in progesterone, androstenedione, or estradiol-17ß concentrations between the groups, testosterone concentrations were slightly lower with the epidural treatment than with the control treatment (p = 0.08). In conclusion, superovulation was induced by a single epidural injection of pFSH, which achieved higher pFSH level than the multiple injections in Holstein dry cows.

Estrus Detection Using Background Image Subtraction Technique in Tie-Stalled Cows.

In this study, we determined the applicability of the background image subtraction technique to detect estrus in tie-stalled cows. To investigate the impact of the camera shooting direction, webcams were set up to capture the front, top, and rear views of a cow simultaneously. Video recording was performed for a total of ten estrous cycles in six cows. Standing estrus was confirmed by testing at 6 h intervals. From the end of estrus, transrectal ultrasonography was performed every 2 h to confirm ovulation time. Foreground objects (moving objects) were extracted in the videos using the background subtraction technique, and the pixels were counted at each frame of five frames-per-second sequences. After calculating the hourly averaged pixel counts, the change in values was expressed as the pixel ratio (total value during the last 24 h/total value during the last 24 to 48 h). The mean pixel ratio gradually increased at approximately 48 h before ovulation, and the highest value was observed at estrus, regardless of the camera shooting direction. When using front-view videos with an appropriate threshold, estrus was detected with 90% sensitivity and 50% precision. The present method in particular has the potential to be a non-contact estrus detection method for tie-stalled cows.

An attempt at estrus detection in cattle by continuous measurements of ventral tail base surface temperature with supervised machine learning.

We aimed to determine the effectiveness of estrus detection based on continuous measurements of the ventral tail base surface temperature (ST) with supervised machine learning in cattle. ST data were obtained through 51 estrus cycles on 11 female cattle (six Holsteins and five Japanese Blacks) using the tail-attached sensor. Three estrus detection models were constructed with the training data (n = 17) using machine learning techniques (random forest, artificial neural network, and support vector machine) based on 13 features extracted from sensing data (indicative of estrus-associated ST changes). Estrus detection abilities of the three models on test data (n = 34) were not statistically different among models in terms of sensitivity and precision (range 50.0% to 58.8% and 60.6% to 73.1%, respectively). The relatively poor performance of the models might indicate the difficulty of separating estrus-associated ST changes from estrus-independent fluctuations in ST.

Production of piglets from in vitro-produced blastocysts by ultrasound-guided ovum pick-up from live donors.

The objective of this research was to develop a system for piglet production by transvaginal ultrasound-guided ovum pick-up (OPU), in vitro production (IVP) of embryos and embryo transfer. First, to establish a culture system for a small number of oocytes or embryos, we evaluated the effect of different incubation volumes and culture densities on fertilizing ability and developmental competence in vitro. Porcine oocytes derived from slaughterhouse ovaries were matured, fertilized and then cultured in vitro in groups as follows: 50 oocytes in 500 µL medium for IVM, 20 oocytes in 100 µL medium for IVF and 20 embryos in 40 µL medium for IVC (Group I); 20 in 100 µL for IVM, 20 in 100 µL for IVF and 20 in 40 µL for IVC (Group II); and 10 in 100 µL for IVM, 10 in 100 µL for IVF and 10 in 40 µL for IVC (Group III). Percentages of sperm penetration, cleavage and blastocyst formation did not differ among the groups. Second, to increase the collection efficiency of porcine oocytes by transvaginal ultrasound-guided OPU, the effects of aspiration pressure on follicular oocyte collection were assessed. Oocytes were aspirated from ovaries of live sows using 80 or 100 mmHg. The recovered oocytes were divided into four categories according to the surrounding cumulus cells and quality of oocytes. The number of oocytes recovered using 100 mmHg pressure was significantly higher than with 80 mmHg pressure. However, there were no significant differences in the population of oocytes grouped by the morphological criteria, number of blastocysts per session and the total cell number in blastocysts between the two vacuum pressures. Finally, 81 oocytes obtained by OPU from five donor sows were subjected to IVP and 47 transferable embryos (9.4 ±â€¯4.0 [mean ±â€¯SD] morulae/blastocysts per session) were obtained at 5 days after IVF. When they were transferred into five recipient gilts (5-16 embryos per recipient), three of five recipients became pregnant and farrowed a total of 12 live piglets. The present results demonstrate that porcine blastocysts can be produced by OPU-IVP and develop to full term after embryo transfer.

Relationships between the antral follicle count, steroidogenesis, and secretion of follicle-stimulating hormone and anti-Müllerian hormone during follicular growth in cattle.

BACKGROUND: The antral follicle count (AFC) in mammalian ovaries positively correlates with female fertility. To clarify the causes of differences in fertility between low and high AFC cows, we investigated follicular growth dynamics and hormone concentrations in plasma, follicular fluid, and in vitro growth (IVG) media at different stages of follicular growth. METHODS: Seven cows were divided into high AFC (n = 4, > 30 follicles) and low AFC (n = 3, < 30 follicles) groups based on the peak AFC detected by ultrasonography. These cows were subjected to estrous synchronization, daily ovarian ultrasonography, and blood collection. Their follicular fluid was collected from dominant follicles at different stages (selection, luteal, and ovulatory phases). In another experiment, we cultured oocyte-cumulus-granulosa cell complexes collected from early antral follicles (< 1 mm) for 12 days. Estradiol-17ß (E2), testosterone (T), progesterone (P4), and anti-Müllerian hormone (AMH) concentrations in follicular fluids and plasma were measured. Plasma follicle-stimulating hormone (FSH) concentrations were examined. E2, P4, and AMH concentrations were also measured in IVG media. RESULTS: The numbers of small (< 4 mm) and intermediate (4-8 mm) follicles were larger in the high AFC group than in the low AFC group (P < 0.05). The number of intermediate follicles was stable in the low AFC group, indicating consistent development. However, the number of these follicles fluctuated in the high AFC group. Plasma FSH concentrations were higher, whereas E2 and T concentrations were lower in the low AFC group (P < 0.05). E2 concentrations and the E2/P4 ratio in ovulatory follicles and IVG media on day 8 were higher in the high AFC group (P < 0.05). AMH concentrations in plasma and IVG media (P < 0.01) were higher in the high AFC group. CONCLUSIONS: The weaker response to FSH of granulosa cells caused low E2 production in the low AFC group, resulting in high FSH concentrations and the consistent development of intermediate follicles. Conversely, higher E2 concentrations suppressed FSH secretion in the high AFC group. Granulosa cells in the high AFC group had the ability to produce more AMH than those in the low AFC group throughout IVG culture.

Estrous detection by continuous measurements of vaginal temperature and conductivity with supervised machine learning in cattle.

This study aimed to evaluate the effectiveness of estrous detection technique based on continuous measurements of vaginal temperature (VT) and conductivity (VC) with supervised machine learning in cattle. The VT and VC of 17 cows in tie-stalls were measured using our developed wearable vaginal sensor from Day 11 (Day 0 = ovulation day) to Day 11 of the subsequent estrous cycle at 15-min interval. After the maximum VT and VC were extracted hourly, their changes were expressed as residual VT (rVT = actual VT - mean VT for the same time on the previous 3 days) and as VC ratio (VCr = actual VC/mean VC for the same time on Day 11-13), respectively, and were used for analysis. Trans-rectal ultrasonography was performed to monitor ovarian structure changes. The plasma concentrations of reproductive hormones (progesterone: P4, estradiol-17ß: E2, and LH) were measured in the experimental period. Standing estrus was confirmed by testing with herd mates at 3-h interval. The rVT decreased transiently, which coincided with decreasing P4 a few days before estrus, and a sharp increase was associated with LH surge during estrus. The VCr increased as estrus approached, corresponding with decreasing P4 and increasing E2 and LH. After noise reduction, features, possible to follow-up estrus-associated changes in rVT and VCr, were extracted and used for developing estrous detection models; 9 models were developed with 3 feature sets (features extracted from rVT alone, VCr alone, and combination of rVT and VCr) and 3 machine learning algorithms (decision tree: DT, support vector machine: SVM, and artificial neural network: ANN). Cross-validation showed that models using the features from the combination of rVT and VCr showed better performance in terms of sensitivity and precision than those using features from VCr alone, and precision than those of using features from rVT alone. Within the models using the features from the combination of rVT and VCr, sensitivity and precision of the model generated by ANN were numerically, but not statistically, higher than those generated by DT and SVM. Of 17 estruses, 16 were detected, with one false positive, when the best model was used. Furthermore, both mean and variance of the interval from the beginning of the estrous detection alert to ovulation (27.3 ±â€¯6.7 h, mean ±â€¯SD of 16 estruses) were not significantly different to those from the onset of standing estrus to ovulation (30.8 ±â€¯5.8 h, n = 17), indicating that the estrus can be detected real-time by the present technique. Hence, the estrous detection technique based on continuous measurements of VT and VC with supervised machine learning has a potential for efficient and accurate estrous detection in cattle.

[Factors influencing parental medication adherence in pediatric asthma].

PURPOSE: To identify the factors influencing parental medication control behaviors (inhaling corticosteroids and medication-taking) in pediatric asthma management. METHODS: A specially-designed questionnaire survey was conducted on 942 parents with asthmatic children in hospitals and elementary schools. RESULTS: Factor analysis on inhalation behaviors resulted in five factors: understanding of benefit, mastering on inhalation skills and medication management, family support, anxiety of side effects, and explanation from a doctor (cumulative contribution ratio=51.3%). Factor analysis on medication-taking behavior resulted in five factors: understanding of medication effectiveness and benefit, family support, anxiety of side effects, skills on giving medicines, and family routine (cumulative contribution ratio=50.6%). CONCLUSION: The results indicate the importance of recognizing factors influencing parental medication control behaviors in developing education strategies to maintain and reinforce their asthma management behaviors.

Evaluation of relative potencies for in vitro transactivation of the baikal seal aryl hydrocarbon receptor by dioxin-like compounds.

To evaluate the sensitivity and responses to dioxins and related compounds (DRCs) via aryl hydrocarbon receptor (AHR) in Baikal seals (Pusa sibirica), we constructed an in vitro reporter gene assay system. Baikal seal AHR (BS AHR) expression plasmid and a reporter plasmid containing CYP1A1 promoter were transfected in COS-7 cells. The cells were treated with six representative congeners, and dose-dependent responses were obtained for all the congeners. EC50 values of 2,3,7,8-TCDD, 1,2,3,7,8-PeCDD, 2,3,7,8-TCDF, 2,3,4,7,8-PeCDF, and PCB126 were found to be 0.021, 1.8, 0.16, 2.4, and 2.5 nM, respectively. As the response did not reach the maximal plateau, EC50 value for PCB118 could not be obtained. The TCDD-EC50 for BS AHR was as high as that for dioxin sensitive C57BL/6 mouse AHR. The in vitro dose responses were further analyzed following an established systematic framework and multiple (20, 50, and 80%) relative potencies (REPs) to the maximum TCDD response. The estimates revealed lower REP ranges (20-80%) of PeCDD and PeCDF for BS AHR than for mouse AHR. Average of the 20, 50, and 80% REPs was designated as Baikal seal specific TCDD induction equivalency factor (BS IEF). The BS IEFs of PeCDD, TCDF, PeCDF, PCB126, and PCB118 were estimated as 0.010, 0.018, 0.0078, 0.0059, and 0.00010, respectively. Total TCDD induction equivalents (IEQs) that were calculated using BS IEFs and hepatic concentrations in wild Baikal seals corresponded to only 12-31% of 2005 WHO TEF-derived TEQs. Nevertheless, about 50% of Baikal seals accumulated IEQs over the TCDD-EC50 obtained in this study. This assessment was supported by the enhanced CYP1A1 mRNA expression found in 50% of the specimens contaminated over the TCDD-EC50. These findings suggest that the IEFs proposed from this in vitro assay could be used to predict AHR-mediated responses in wild seals.

[Prospective observational study of clinical course after stopping inhaled corticosteroids in child asthma].

BACKGROUND: Prospective study was carried out to observe the clinical course and find out the clinical index to predict the outcome after stopping of inhaled corticosteroids (ICS). SUBJECTS: One hundred twelve asthmatic children (aged 12.9+/-9.6 years old) who were totally controlled with 100 microg/day of inhaled corticosteroids and have not experienced asthma symptom over 3 months, were enrolled. METHODS: Patients with over PC20 0.5 mg/mL of methacholine challenge test (MCT) were recommended to stop ICS therapy. Clinical course, lung function and bronchial hyperreactivity were followed at 7, 16 and 24 months after stopping of ICS. RESULTS: Eighty-one percent of 63 patients followed up to 24 months run on as symptom less than one time per month without remarkable change of lung function. PC20 in group of no asthma symptom during 24 months (group A) decreased weakly. That of other groups decreased remarkably. Days using ICS and plasma IgE antibody at stopping of ICS were different in clinical course at 24 months after stopping ICS. CONCLUSION: Over 80% of totally controlled asthmatic children with MCT PC20 over 0.5 mg/mL could stop ICS therapy and run on without relapse.

[Clinical features of severe infant atopic dermatitis with hypoproteinemia].

OBJECTIVE: In recent years, there have been sporadic reports of severe atopic dermatitis (AD) with hypoproteinemia and growth impairment. The present study was conduced in order to ascertain the characteristics of patients with severe infant AD with hypoproteinemia at the initial visit and their treatment courses. SUBJECTS: Of AD patients younger than 1 year of age who visited the department over a 27-month period from March 2002, subjects were those with severe AD accompanied by hypoproteinemia. The clinical characteristics of these patients at the initial visit and the changes in symptoms and laboratory findings were statistically analyzed. RESULTS: Of the total of 119 AD patients younger than 1 year of age visited to the department during the above-mentioned period, 15 patients had severe AD with hypoproteinemia. The height and body weight of approximately half of the patients were less than 3rd percentile, and 10% and more of the patients had severe hyponatremia or hyperpotassemia. The platelet count for 60% of the patients exceeded 800 x 10(3)/microl. After visiting the department, therapy involving the use of skin care products and topical steroids and the removal of exacerbation factors quickly improved dermal symptoms and laboratory findings. CONCLUSION: Severe AD is a disease that should be cautiously treated because of the risk of hypoproteinemia, growth impairment, electrolyte abnormalities, and thrombocytosis; however, it should be noted that appropriate treatments can improve this condition.

Aryl hydrocarbon receptor (AHR) and AHR nuclear translocator (ARNT) expression in Baikal seal (Pusa sibirica) and association with 2,3,7,8-TCDD toxic equivalents and CYP1 expression levels.

Most toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related planar halogenated aromatic hydrocarbons (PHAHs) are mediated by ligand-activated aryl hydrocarbon receptor (AHR) signaling pathway. To understand the regulation mechanism of AHR and AHR nuclear translocator (ARNT) expression in wild Baikal seal (Pusa sibirica) population contaminated by PHAHs, the present study investigated hepatic mRNA expression levels of AHR and its heterodimer, ARNT genes, in association with biological index (age, gender and body weight), PHAH accumulation and expression levels of cytochrome P450 (CYP) 1A and 1B. While there was no gender difference, the AHR mRNA expression levels were increased with ages (p = 0.014) and body weights (p = 0.015), indicating that AHR expression might be affected by these biological factors. The AHR mRNA expression levels exhibited significant positive correlations with total TEQs and most of individual congener TEQs derived from polychorinated dibenzo-p-dioxins, dibenzofurans and non-ortho coplanar polychorinated biphenyls (PCBs), indicating the transcriptional up-regulation of AHR expression by these congeners. On the other hand, there was no significant correlation between individual TEQs from mono-ortho coplanar PCBs and AHR expression. These results imply the structure-related transcriptional activity of AHR among PHAHs congeners. AHR mRNA levels showed positive correlations with both CYP1A protein (p = 0.039) and CYP1A1 mRNA expression levels (p = 0.046). In contrast to AHR expression, neither the total nor individual congener TEQs influenced ARNT at the transcriptional level. ARNT mRNA showed significant negative correlations with CYP1A/1B protein (p = 0.027 and p = 0.006) and CYP1A1 mRNA expression levels (p = 0.039), implying the existence of different transcriptional regulation between AHR and ARNT genes and negative regulation by CYP1A/1B-mediated signaling pathways. The present findings may render significant insight on the basic mechanisms underlying regulation of AHR and ARNT expressions associated with biological factors and PHAH exposure in wild mammalian populations.

Analysis of gene expressions of T cells from children with acute exacerbations of asthma.

BACKGROUND: Little is known about airway inflammation in childhood asthma. The aims of this study were to analyze the expression of a wide range of mediators of airway inflammation in childhood asthma. METHODS: Eight asthmatic children with acute exacerbations were recruited for the study. Peripheral blood was drawn from the patients at the time of exacerbation and after improvement. Total RNA was extracted from the isolated T lymphocytes. The differential display (DD) RT-PCR method was used to detect expressed genes, and the quantification of candidate gene expression was performed using real-time quantitative RT-PCR. RESULTS: The only gene for which significant expression differences were detected in both DD analysis and quantitative RT-PCR was lipocortin II (annexin II) (exacerbation > remission, p < 0.05). In quantitative RT-PCR of interleukin-4 (IL-4), IL-5, interferon-gamma, IL-12 receptor-beta and integrin alpha6, a significant difference was found only in the expression of IL-4 mRNA (exacerbation > remission, p < 0.05). The IL-4 plasma concentration tended to be higher in exacerbation than in remission. CONCLUSIONS: Our findings suggest activation of T cells and IL-4 production may be involved not only in the basic pathogenesis of childhood asthma but also in its acute exacerbation, and that lipocortin II may be a marker or contribute to asthma exacerbation.