Association of intrauterine presence of Lactobacillus spp. with inflammation and pathogenic bacteria in the uterus in postpartum dairy cows.

The aim of this study was to clarify the influence of Lactobacillus spp. on the degree of endometrial inflammation in the postpartum period and the relationship between Lactobacillus spp. and pathogenic bacteria in the endometrium of postpartum dairy cows. Endometrial samples were collected from 41 Holstein-Friesian cows at 4 and 8 weeks postpartum using cytobrushes for polymorphonuclear neutrophil (PMN) count and bacterial culture to isolate Lactobacillus spp., Escherichia coli, and Trueperella pyogenes. The 4-week samples were divided into four groups (E+L+), (E+L-), (E-L+), (E-L-) according to whether endometritis was diagnosed (E+) and Lactobacillus spp. was isolated (L+). The diagnostic criterion for cytological endometritis was > 18% PMN. The average PMN% in the E+L+ group was lower than that in the E+L-group (P < 0.05) at 8 weeks postpartum. There were no significant correlations between the number of colonies of Lactobacillus spp. and E. coli or between that of Lactobacillus spp. and T. pyogenes. Lactobacillus spp. could reduce PMN% in dairy cows with endometritis during the puerperal period. In conclusion, the intrauterine presence of Lactobacillus spp. may have a positive effect on uterine involution in postpartum dairy cows.

Cardiovascular magnetic resonance feature tracking for characterization of patients with heart failure with preserved ejection fraction: correlation of global longitudinal strain with invasive diastolic functional indices.

BACKGROUND: Left ventricular (LV) diastolic dysfunction is the main cause of heart failure with preserved ejection fraction (HFpEF), and is characterized by LV stiffness and relaxation. Abnormal LV global longitudinal strain (GLS) is frequently observed l in HFpEF, and was shown to be useful in identifying HFpEF patients at high risk for a cardiovascular event. Cardiovascular magnetic resonance (CMR) feature tracking (CMR-FT) enables the reproducible and non-invasive assessment of global strain from cine CMR images. However, the association between GLS and invasively measured parameters of diastolic function has not been investigated. We sought to determine the prevalence and severity of GLS impairment in patients with HFpEF by using CMR-FT, and to evaluate the correlation between GLS measured by CMR-FT and that measured by invasive diastolic functional indices. METHODS: Eighteen patients with HFpEF and 18 age- and sex-matched healthy control subjects were studied. All subjects underwent cine, pre- and post-contrast T1 mapping and late gadolinium-enhancement CMR. In the HFpEF patients, invasive pressure-volume loops were obtained to evaluate LV diastolic properties. GLS was quantified from cine CMR, and extracellular volume fraction (ECV) was quantified from pre- and post-contrast T1 mapping as a known imaging biomarker for predicting LV stiffness. RESULTS: GLS was significantly impaired in patients with HFpEF (- 14.8 ± 3.3 vs.-19.5 ± 2.8%, p < 0.001). Thirty nine percent (7/18) of HFpEF patients showed impaired GLS with a cut-off of - 13.9%. Statistically significant difference was found in ECV between HFpEF patients and controls (32.2 ± 3.8% vs. 29.9 ± 2.6%, p = 0.044). In HFpEF patients, the time constant of active LV relaxation (Tau) was strongly correlated with GLS (r = 0.817, p < 0.001), global circumferential strain (GCS) (r = 0.539, p = 0.021) and global radial strain (GRS) (r = - 0.552, p = 0.017). Multiple linear regression analysis revealed GLS as the only independent predictor of altered Tau (beta = 0.817, p < 0.001) among age, LV end-diastolic volume index, LV end-systolic volume index, LV mass index, GCS, GRS and GLS. CONCLUSIONS: CMR-FT is a noninvasive approach that enables identification of the subgroup of HFpEF patients with impaired GLS. CMR LV GLS independently predicts abnormal invasive LV relaxation index Tau measurements in HFpEF patients. These findings suggest that feature-tracking CMR analysis in conjunction with ECV, may enable evaluation of diastolic dysfunction in patients with HFpEF.

Real-time decision-making during emergency disease outbreaks.

In the event of a new infectious disease outbreak, mathematical and simulation models are commonly used to inform policy by evaluating which control strategies will minimize the impact of the epidemic. In the early stages of such outbreaks, substantial parameter uncertainty may limit the ability of models to provide accurate predictions, and policymakers do not have the luxury of waiting for data to alleviate this state of uncertainty. For policymakers, however, it is the selection of the optimal control intervention in the face of uncertainty, rather than accuracy of model predictions, that is the measure of success that counts. We simulate the process of real-time decision-making by fitting an epidemic model to observed, spatially-explicit, infection data at weekly intervals throughout two historical outbreaks of foot-and-mouth disease, UK in 2001 and Miyazaki, Japan in 2010, and compare forward simulations of the impact of switching to an alternative control intervention at the time point in question. These are compared to policy recommendations generated in hindsight using data from the entire outbreak, thereby comparing the best we could have done at the time with the best we could have done in retrospect. Our results show that the control policy that would have been chosen using all the data is also identified from an early stage in an outbreak using only the available data, despite high variability in projections of epidemic size. Critically, we find that it is an improved understanding of the locations of infected farms, rather than improved estimates of transmission parameters, that drives improved prediction of the relative performance of control interventions. However, the ability to estimate undetected infectious premises is a function of uncertainty in the transmission parameters. Here, we demonstrate the need for both real-time model fitting and generating projections to evaluate alternative control interventions throughout an outbreak. Our results highlight the use of using models at outbreak onset to inform policy and the importance of state-dependent interventions that adapt in response to additional information throughout an outbreak.

Extended-spectrum ß-lactamase-producing Escherichia coli isolated from healthy Thoroughbred racehorses in Japan.

Extended-spectrum ß-lactamase-producing Escherichia coli (ESBLEC) have become a major health concern in both human and veterinary medicine. These bacteria could become a critical problem in equine medicine due to the limited number of antimicrobial drugs available. However, there are no previous reports of ESBLEC isolated from horses in Japan. The objectives of this study were to investigate the occurrence of ESBLEC isolated from feces in healthy Thoroughbred racehorses in Japan. Feces samples were collected from 147 healthy Thoroughbred racehorses by equine veterinarians at the Japan Racing Association (103 from Miho Training Center and 44 from Ritto Training Center) between March 2017 and April 2018. Samples were screened for ESBLECs using MacConkey agar supplemented with 1 µg/ml cefotaxime. Detection of ESBL genes was performed by PCR and confirmed by DNA sequencing. Horizontal transmission was demonstrated by conjugation assay. In this study, 24 ESBLECs were isolated from twelve horse feces samples (8.2%). All ESBLECs harbored blaCTX-M-2, and both blaTEM-1 and blaCTX-M-2 were detected in nine isolates (37.5%). ESBLECs showed resistance to all ß-lactam antibiotics (100%) tested, followed by trimethoprim (66.7%), streptomycin (62.5%), tetracycline (25.0%), and oxytetracycline (25.0%). Horizontal transmission was successfully demonstrated by conjugation assay in eight of 13 isolates, and blaCTX-M-2 was detected by PCR in all transconjugants. This study showed that racehorses in Japan are potential reservoirs of ESBLECs.

Diagnostic Accuracy of Endocardial-to-Epicardial Myocardial Blood Flow Ratio for the Detection of Significant Coronary Artery Disease With Dynamic Myocardial Perfusion Dual-Source Computed Tomography.

BACKGROUND: Previous dynamic stress computed tomography perfusion (CTP) studies used absolute myocardial blood flow (MBF in mL/100 g/min) as a threshold to discriminate flow-limiting coronary artery disease (CAD), but absolute MBF can be vary because of multiple factors. The aim of this study was to compare the diagnostic performance of absolute MBF and the transmural perfusion ratio (TPR) for the detection of flow-limiting CAD, and to clarify the influence of CT delayed enhancement (CTDE) on the diagnostic performance of CTP.Methods and Results:We retrospectively enrolled 51 patients who underwent dual-source CTP and invasive coronary angiography (ICA). TPR was defined as the endocardial MBF of a specific segment divided by the mean of the epicardial MBF of all segments. Flow-limiting CAD was defined as luminal diameter stenosis >90% on ICA or a lesion with fractional flow reserve ≤0.8. Segmental presence and absence of myocardial scar was determined by CTDE. The area under the receiver-operating characteristics curve (AUC) of TPR was significantly greater than that of MBF for the detection of flow-limiting CAD (0.833 vs. 0.711, P=0.0273). Myocardial DE was present in 27 of the 51 patients and in 34 of 143 territories. When only territories containing DE were considered, the AUC of TPR decreased to 0.733. CONCLUSIONS: TPR calculated from absolute MBF demonstrated higher diagnostic performance for the discrimination of flow-limiting CAD when compared with absolute MBF itself.

Disagreement between parents on assessment of child temperament traits.

BACKGROUND: Accuracy of temperament assessment is a prerequisite in research studies. To identify the extent to which parental assessment of child temperament is biased by their personal attributes, we proposed a new structural equation model, in which biases of parental attributes in their assessment of child temperament can be separated from the true (i.e. non-biased) associations between the two. METHODS: We examined 234 father-mother pairs using questionnaires including Emotionality, Activity, Sociability, and Impulsivity; Social Desirability Scale; Hospital Anxiety and Depression Scale; Temperament and Character Inventory; and State-Trait Anger Expression Inventory. RESULTS: Paternal Depression and Persistence, maternal Trait Anger, and parental Novelty Seeking showed significant bias in assessment of Emotionality. Maternal Self-transcendence showed significant bias in assessment of child Impulsivity. CONCLUSION: Researchers should be cautious about biases in parental assessment of children's Emotionality and Impulsivity, but other temperament traits may be free from such biases.

Comparative analysis of LTR and structural genes in an equine infectious anemia virus strain isolated from a feral horse in Japan.

Although equine infectious anemia virus (EIAV) poses a major threat to the equine industry worldwide, the molecular epidemiology of this virus is poorly understood. Recently, an EIAV strain (EIAVMiyazaki2011-A) representing a new monophyletic group was discovered in feral horses in southern Japan. In the present study, the EIAVMiyazaki2011-A proviral genome is compared with evolutionarily divergent EIAV isolates to investigate conservation of functional elements or motifs within the long terminal repeats (LTRs) and structural genes. This analysis represents a significant step forward in increasing understanding of the molecular conservation and variation between geographically distinct strains of this equine lentivirus.

Bos grunniens papillomavirus type 1: a novel deltapapillomavirus associated with fibropapilloma in yak.

Papillomaviruses (PVs) have been widely identified among vertebrates, but have not yet been reported to infect yaks. We report, for the first time, a novel deltapapillomavirus that was associated with fibropapilloma in yak herds on the Qinghai-Tibetan Plateau. Six skin papilloma samples were collected and examined using histopathology, immunohistochemistry and PCR assays. The samples were identified as fibropapilloma and were found to contain PV antigens. Sequencing of diagnostic PCR products and the full-length genome revealed that all samples were infected with the same PV type. The whole virus genome was 7946 bp in length and possessed the common PV genomic organization. The virus was identified as a novel PV type and designated Bos grunniens papillomavirus type 1 (BgPV-1) based on the nucleotide sequence alignment of the L1 ORF. It is classified in the Delta-4 species of the genus Deltapapillomavirus based on phylogenetic analysis of the L1 ORF. Identification of this novel PV type provides further information about the pathology, development of diagnostic methods and evolutionary studies of the family Papillomaviridae.

Identification of a novel equine infectious anemia virus field strain isolated from feral horses in southern Japan.

Although equine infectious anemia (EIA) was described more than 150 years ago, complete genomic sequences have only been obtained from two field strains of EIA virus (EIAV), EIAV(Wyoming) and EIAV(Liaoning). In 2011, EIA was detected within the distinctive feral Misaki horse population that inhabits the Toi-Cape area of southern Japan. Complete proviral sequences comprising a novel field strain were amplified directly from peripheral blood of one of these EIAV-infected horses and characterized by nucleotide sequencing. The complete provirus of Miyazaki2011-A strain is 8208 bp in length with an overall genomic organization typical of EIAV. However, this field isolate possesses just 77.2 and 78.7 % nucleotide sequence identity with the EIAV(Wyoming) and EIAV(Liaoning) strains, respectively, while similarity plot analysis suggested all three strains arose independently. Furthermore, phylogenetic studies using sequences obtained from all EIAV-infected Misaki horses against known viral strains strongly suggests these Japanese isolates comprise a separate monophyletic group.

Characterization of novel bovine papillomavirus type 12 (BPV-12) causing epithelial papilloma.

Bovine papillomavirus type 12 (BPV-12, putative type BAA1) was detected in epithelial papilloma located on the tongue of an infected cow. Then, the whole genome was sequenced, and phylogenetic analysis illustrated that it should be classified as a member of the genus Xipapillomavirus. The viral genome is 7197 base pairs in length and contains five early ORFs (E1, E2, E4, E7 and E8), three late ORFs (L1, L2 and L3), and a long control region that possesses replication regulatory elements. Meanwhile, mRNA of each gene was detected in the papilloma sample. The papilloma was identified as epithelial papilloma by histological and immunohistochemical examination. Based on the genome information and pathological properties, BAA1 was designated as BPV-12 in this study.

Development of a nested PCR assay to detect equine infectious anemia proviral DNA from peripheral blood of naturally infected horses.

Equine infectious anemia (EIA) has posed a major challenge and caused significant losses to the equine industry worldwide. PCR detection methods have considerable potential as an adjunct to conventional serological diagnostic techniques. However, most published PCR methods, including that recommended by the OIE, were designed using laboratory-adapted virus strains and do not function with field isolates of EIA virus (EIAV). In the present study, a nested PCR assay for detection of EIAV proviral DNA in peripheral blood cells of naturally infected horses was developed. Primer sets were designed based on conserved 5' regions of the viral genome extending from the LTR to the tat gene. Preliminary studies demonstrated that the method has a detection limit of 10 genomic copies and, when applied to a naturally EIAV-infected feral horse population, shows 100 % correlation with conventional serological diagnostic techniques. This assay provides a powerful new tool in the control of EIAV.

Single application of A2 NTX, a botulinum toxin A2 subunit, prevents chronic pain over long periods in both diabetic and spinal cord injury-induced neuropathic pain models.

Botulinum toxin type A is a unique candidate for inhibition of pain transmission. In the present study we attempted to see the beneficial actions of A2 neurotoxin (NTX), an active subunit of botulinum toxin type A. Intraplantar injection of A2 NTX significantly suppressed mechanical allodynia and hypersensitivities to A-fiber stimuli in the diabetic neuropathic pain model. Spinal application of A2 NTX also showed a potent suppression of thermal hyperalgesia and mechanical allodynia in the spinal cord injury-induced neuropathic pain model. A2 NTX seems to be a long-lasting treatment for diabetic and spinal cord injury-induced neuropathic pain.

Intergenerational transmission of child-rearing styles mediated by adult attachment: A structural equation model approach.

Aim: Parenting styles are determinants of psychosocial maladjustment in the subsequent generation. Modifiable factors that positively impact intergenerational transmission should be identified. Thus, this study investigated the mediating effects of adult attachment on the transmission of child-rearing styles and the combined impact of fathers' and mothers' styles on their offspring's parenting. Methods: Overall, 396 fathers and 733 mothers participated. They were married or cohabiting when their children aged 0-10 years attended a pediatric office in Japan. The pediatricians distributed the self-report questionnaires to examine perceived rearing (the Parental Bonding Instrument), adult attachment (the Relationship Questionnaire), and demographic characteristics. We then analyzed the data using structural equation modeling that illustrated intergenerational transmission of rearing styles between grandparents and parents. Results: Overall, 385 fathers (M age = 35.5 years, SD = 6.0) and 699 mothers (M age = 33.5 years, SD = 5.1) satisfied the eligibility criteria. The path models showed that adult attachment to partners mediated grandparental and parental care. However, no mediating effect was observed in other intergenerational patterns that transmitted overprotective rearing styles. Additionally, combined grandfather and grandmother care were positively associated with parental overprotection. Conclusion: This study suggested the potential of adult attachment in mediating loving, empathic, and warm rearing styles and the combined effects of both parents' styles on promoting the next-generation parents' overprotective styles. Our findings clarify how to terminate the negative chain of the parent-child transmission.

Usefulness of a new DUV-LED device for the control of infection by Escherichia coli, Staphylococcus aureus, mycobacteria and spore-forming bacteria.

Reliable disinfection and sterilization technologies are needed to deal with the various infectious diseases spreading around the world. Furthermore, bacteria that are difficult to eliminate by ordinary disinfection are also a problem in the medical environment. We examined the germicidal effect of a newly developed deep-ultraviolet light-emitting diode (DUV-LED) prototype device (wavelength of 280 ± 5 nm; power of 0.9 to 1.4 mW/cm2) for floor sterilization against Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), Mycobacterium gordonae (M. gordonae), and Bacillus subtilis (B. subtilis). This prototype device is equipped with highly practical DUV-LEDs with a high output efficiency and a long life, and was designed with consideration of the irradiation distance and the angle of the DUV-LEDs to provide a uniform irradiation rate. We found a statistically significant reduction of ≥90% in the infectious titers of both E. coli and S. aureus after irradiation for 2 s. Although acid-fast bacilli and spore-type bacilli are generally thought to be resistant to UV light irradiation compared to general bacteria, the acid-fast bacillus M. gordonae was inactivated after irradiation for 10 s, and spore-type cells of the bacillus B. subtilis were inactivated by ≥90% after irradiation for 30 s. We also found that the effects were cumulative when irradiation was performed at intervals. In the future, the usefulness of this device as an infection control measure will be evaluated in daily medical practice.

Late catch-up phenomenon associated with stent fracture after sirolimus-eluting stent implantation: incidence and outcome.

OBJECTIVE: To examine the long-term outcome of the stent fracture (SF) and the potential predictive factors contributing to in-stent restenosis (ISR) in the fractured stent. BACKGROUND: The SF is thought to be a higher risk of ISR in drug-eluting stent, although SF does not always develop ISR. METHODS: The consecutive 1,228 de novo lesions in 1,079 patients who underwent sirolimus-eluting stents implantation and assessed by 8 months follow-up coronary angiography were retrospectively analyzed. RESULTS: One hundred and seventeen SFs (9.5%) were identified in 100 patients and 22 (18.8%) SFs revealed ISR at the first follow-up. In addition, 16 (13.7%) developed new ISRs from 95 residual SFs without ISR prior to the second follow-up. Overall, 38 (32.5%) of all 117 SFs developed ISR, and 16 (42.1%) of 38 SFs occurred in a late phase beyond the first 8 months follow-up. A higher risk of ISR in the SF site was associated with the chronic total occlusion (ISR vs. no ISR: 34.2% vs. 16.5%, P = 0.0304), calcified lesions (55.3% vs. 34.2%, P = 0.0299), and correspondence 89.5% versus 43.0%, P < 0.0001 (SF site occurring at the original target lesion site) in the univariate analysis. The correspondence was identified as the only strong predictive factor for ISR at the SF site according to a multivariate logistic regression analysis (odds ratio 12.6, 95% confidence interval 3.82-53.5, P < 0.0001). CONCLUSIONS: SF occurring at the site of the original target lesion was a strong independent predictor of ISR. This indicates the need for a careful, long-term follow-up in those situations, even when no significant ISR is initially detected.

Type A1 but not type A2 botulinum toxin decreases the grip strength of the contralateral foreleg through axonal transport from the toxin-treated foreleg of rats.

The adverse effects of botulinum LL toxin and neurotoxin produced by subtype A1 (A1LL and A1NTX) are becoming issues, as the toxins could diffuse from the toxin-treated (ipsilateral) to contralateral muscles. We have attempted to produce neurotoxin from subtype A2 (A2NTX) with an amino acid sequence different from that of neurotoxin subtype A1. We measured the grip strength on the contralateral foreleg as an indicator of toxin spread from the ipsilateral to contralateral muscles. Doses of 0.30 log U or above of A1LL and A1NTX reduced the contralateral grip strength, whereas a dose of 0.78 log U of A2NTX was required to do so. We investigated the route of toxin spread using denervated, colchicine-treated, and antitoxin-treated rats. A1LL was transported via axons at doses higher than 0.30 log U and via both axons and body fluid at about 0.80 log U or a higher dose. Interestingly, A2NTX was transported via body fluid at about 0.80 log U or a higher dose, but not via axons to the contralateral side. It was concluded that A1LL and A1NTX decreased the grip strength of the toxin-untreated foreleg via both axonal transport and body fluids, while A2NTX was only transported via the body fluid.

Rapid and sensitive detection of bovine coronavirus and group a bovine rotavirus from fecal samples by using one-step duplex RT-PCR assay.

Bovine coronavirus (BCoV) and group A bovine rotavirus (BRV) are two of major causes for neonatal calf diarrhea. In the present study, a one-step duplex RT-PCR was established to detect and differentiate BCoV and group A BRV from fecal samples. The sensitivity of this method for BCoV and group A BRV was 10 PFU/100 µl and 1 PFU/100 µl, respectively. Twenty-eight diarrhea fecal samples were detected with this method, the result showed that 2 samples were identified as co-infected with BCoV and group A BRV, 26 samples were group A BRV positive, and 2 samples were negative. It proved that this method is sensitive for clinical fecal samples and is worth applying to laboratory diagnosis for BCoV and group A BRV.

Suppurative necrotizing bronchopneumonia caused by Nocardia cyriacigeorgica infection in a stranded striped dolphin (Stenella coeruleoalba) in Japan.

On a coastline in Miyazaki Prefecture, Japan, a wild subadult female striped dolphin was found dead. Necropsy revealed poor nutritional status and bilateral pneumonia, which was histologically diagnosed as severe suppurative necrotizing bronchopneumonia. Special staining detected numerous intralesional filamentous, branching bacteria, which was identified as Nocardia cyriacigeorgica by sequencing of 16S ribosomal RNA and gyrB genes. Other main histological findings included lymphoid depletion in the spleen and superficial cervical and pulmonary lymph nodes. Suppurative nocardiosis without a granulomatous reaction is uncommon, and it is assumed its pathogenesis was related to the host's immune status. This paper discusses the variable inflammatory response to nocardiosis and describes the first case of N. cyriacigeorgica infection in a wild striped dolphin in Japan.

Evaluation of a booster tuberculosis vaccine containing mycobacterial DNA-binding protein 1 and CpG oligodeoxynucleotide G9.1 using a Guinea pig model that elicits immunity to Bacillus Calmette-Guérin.

Tuberculosis is a major threat to global health and its increased incidence in adolescents as well as onset in the elderly presents a serious problem. One strategy to control tuberculosis involves taking advantage of Bacillus Calmette-Guérin's (BCG) superior effects on childhood tuberculosis. Accordingly, here we aimed to develop a booster vaccine for adults who received the BCG vaccine during early childhood. Therefore, we first devised a system to assess the efficacy of a candidate booster vaccine. Specifically, variant strain BCG-II, a minor component of BCG-Tokyo strain, which elicits weak immunity, was administered to guinea pigs. Vaccine-induced immunity and protection against Mycobacterium tuberculosis (Mtb) infection were evaluated using skin delayed-type hypersensitivity (DTH) and Mtb colony forming unit counts in organs, respectively. Candidate booster vaccine containing the mycobacterial DNA-binding protein 1 (MDP1) as antigen and CpG oligodeoxynucleotide G9.1 as adjuvant increased T-bet expression and IFN-γ production in human peripheral blood mononuclear cells. Intradermal administration of MDP1 or MDP1 and G9.1 to unimmunized guinea pigs produced DTH on MDP1-inoculated skin. Boosting BCG-II-primed guinea pigs with this protocol effectively enhanced DTH against MDP1 and protection against Mtb infection, particularly when combined with G9.1. The candidate vaccine may contribute to efforts to prevent tuberculosis.