mRNA expression of immune factors by milk somatic cells from healthy Holstein lactating cows.

This study investigated the mRNA of immune factors expressed by milk somatic cells from 72 healthy lactating Holstein cows on 1 farm. Milk samples were collected aseptically from the right front mammary gland before milking. The milk samples that had a negative reaction to the California mastitis test were used to analyze the mRNA of immune factors. Cows were divided into 2 groups based on the detection of bacteria in milk samples: positive group (n = 22 cows), which showed bacteria in cultures, and negative group (n = 50 cows), which did not show bacteria in cultures. There were significant positive correlations among the relative mRNA levels of interleukin (IL)-6, IL-8, arginase 1, chemokine (C-C motif) ligand (CCL) 1, and chemokine (C-X-C motif) ligand (CXCL) 13, as well as among the relative mRNA levels of IL-10, pentraxin 3, CCL5, and CCL14. Significantly high levels of IL-1ß, IL-6, IL-8, arginase 1, Batf, CCL1, CXCL14, and toll-like receptor 4 in the positive group were discovered compared to the negative group. These results suggest that the presence of bacteria in lactating healthy dairy cows may affect mRNA levels of inflammatory mediators expressed by somatic cells.

Cette étude a examiné l'ARNm des facteurs immunitaires exprimés par les cellules somatiques du lait de 72 vaches Holstein en lactation en bonne santé dans une ferme. Des échantillons de lait ont été prélevés aseptiquement du quartier avant droit de la glande mammaire avant la traite. Les échantillons de lait ayant eu une réaction négative au test de mammite de Californie ont été utilisés pour analyser l'ARNm des facteurs immunitaires. Les vaches ont été divisées en deux groupes sur la base de la détection de bactéries dans les échantillons de lait : groupe positif (n = 22 vaches), qui a montré la présence de bactéries lors des cultures, et groupe négatif (n = 50 vaches), qui n'a pas montré de bactéries lors des cultures. Il y avait des corrélations positives significatives entre les niveaux relatifs d'ARNm de l'interleukine (IL)-6, de l'IL-8, de l'arginase 1, du ligand de chimiokine (motif C-C) (CCL) 1 et du ligand de chimiokine (motif C-X-C) (CXCL) 13, ainsi que parmi les niveaux relatifs d'ARNm d'IL-10, de pentraxine 3, de CCL5 et de CCL14. Des niveaux significativement élevés d'IL-1ß, d'IL-6, d'IL-8, d'arginase 1, de Batf, de CCL1, de CXCL14 et de récepteur de type Toll 4 dans le groupe positif ont été découverts par rapport au groupe négatif. Ces résultats suggèrent que la présence de bactéries chez des vaches laitières saines en lactation peut affecter les niveaux d'ARNm des médiateurs inflammatoires exprimés par les cellules somatiques.(Traduit par Docteur Serge Messier).

Whole-Genome Sequencing of Pasteurella multocida Strain Pm1, Isolated from a Calf.

Bovine pneumonia is a disease that causes significant economic losses in livestock industries and is vital for animal welfare. The whole-genome sequence of Pasteurella multocida strain Pm1, isolated from a calf suffering from pneumonia in Japan, is reported here.

Intra-macrophage expression of ArtAB toxin gene in Salmonella.

Salmonella enterica subspecies enterica serovar Typhimurium (S. Typhimurium) definitive phage type 104 (DT104), S. Worthington, and S. bongori produce ArtAB toxin, which catalyses ADP-ribosylation of pertussis toxin-sensitive G protein. ArtAB gene (artAB) is encoded on a prophage in Salmonella, and prophage induction by SOS-inducing agents is associated with increases in ArtAB production in vitro. However, little is known about the expression of artAB in vivo. Here, we showed a significant increase in artAB transcription of DT104 within macrophage-like RAW264.7 cells. Intracellular expression of ArtAB was also observed by immunofluorescence staining. The induced expression of artAB in DT104 and S. bongori was enhanced by treatment of RAW264.7 cells with phorbol 12-myristate 13-acetate (PMA), which stimulates the production of reactive oxygen species (ROS); however, such induction was not observed in S. Worthington. Upregulation of oxyR, a major regulator of oxidative stress, and cI, a repressor of prophage induction, was observed in S. Worthington within RAW264.7 cells treated with PMA but not in the DT104 strain. Although the expression of oxyR was increased, artAB was upregulated in S. bongori, which lacks the cI gene in the incomplete artAB-encoded prophage. Taken together, oxidative stress plays a role in the production of artAB toxins in macrophages, and high expression levels of oxyR and cI are responsible for the low expression of artAB. Therefore, strain variation in the level of artAB expression within macrophages could be explained by differences in the oxidative stress response of bacteria and might be reflected in its virulence.

Relationship between Escherichia coli virulence factors, notably kpsMTII, and symptoms of clinical metritis and endometritis in dairy cows.

Although Escherichia coli is a commensal bacterium of the bovine vaginal microbiota, it is an important pathogenic bacterium that causes diseases of the reproductive tract and sub-fertility. Recent studies have focused on virulence factors (VFs) of intrauterine E. coli; however, actual endometrial VFs have not been clearly identified. The purpose of this study was to identify the VFs of E. coli associated with clinical metritis and endometritis. Thirty-two strains of E. coli and four mixed Trueperella pyogenes (TP) strains were detected in the uterus of 19 Holstein dairy cows with obvious clinical signs (between 8 and 66 days postpartum). The presence of six E. coli VFs (fimH, fyuA, kpsMTII, hra1, csgA, and astA) was examined by PCR, and clinical signs and reproductive performance (mixed TP, the percentage of polymorphonuclear neutrophils [PMN%], days to uterine involution, etc.) were evaluated. Four VFs (fimH, hra1, csgA, and astA) were detected in all E. coli strains, whereas fyuA and kpsMTII were detected in 94% and 50% of strains, respectively. Cows with E. coli strains harboring kpsMTII exhibited significantly severe clinical scores (vaginal discharge score, PMN%, uterine involution), suggesting that kpsMTII is a key VF for progression of clinical metritis and endometritis. In the present study, we clearly identified six VFs associated with clinical metritis and endometritis. In addition, E. coli strains with kpsMTII probably play a crucial role in the progression of clinical metritis and endometritis.

SOMO-HOMO Conversion in Triplet Cyclopentane-1,3-diyl Diradicals.

According to the Aufbau principle, singly occupied molecular orbitals (SOMOs) are energetically higher lying than a highest doubly occupied molecular orbital (HOMO) in the electronically ground state of radicals. However, in the last decade, SOMO-HOMO-converted species have been reported in a limited group of radicals, such as distonic anion radicals and nitroxides. In this study, SOMO-HOMO conversion was observed in triplet 2,2-difluorocyclopentane-1,3-diyl diradicals DR3F1, DR4F1, and 2-fluorocyclopentante-1,3-diyl diradical DR3HF1, which contain the anthracyl unit at the remote position. The high HOMO energy in the anthracyl moiety and the low-lying SOMO-1 due to the fluoro-substituent effect are the key to the SOMO-HOMO conversion phenomenon. Furthermore, the cation radical DR3F1 + generated through the one-electron oxidation of DR3F1 was found to be a SOMO-HOMO-converted monoradical.

Dispersion of Complex Refractive Indices for Intense Vibrational Bands. I. Quantitative Spectra.

This paper reviews the dispersion of complex refractive indices for representative intense vibrational bands of organic liquids in the absolute scale. Conspicuous variations in both real and imaginary refractive indices over the intense bands are precisely determined as a function of infrared wavenumber by attenuated total reflection infrared absorption measurement. The accurate spectral data offer an excellent reference to calibrate the absolute infrared intensities by various quantum chemical calculations, and thus, critical comparison between the present experiment and computation was reported to clarify the general accuracy of the quantum chemical calculations. The precise data of the dispersion will be utilized to clarify their impact on the analysis of vibrational spectroscopy of interfaces in the subsequent paper.

Dispersion of Complex Refractive Indices for Intense Vibrational Bands. II. Implication to Sum Frequency Generation Spectroscopy.

Sharp and intense vibrational bands are characterized with conspicuous dispersion of complex refractive indices. Based on the quantitative data of dispersion in the preceding paper, this paper clarifies the influence of the dispersion on the sum frequency generation (SFG) spectroscopy. As a consequence of the large dispersion, the lineshapes of SFG spectra could be influenced by the frequency dependence of the Fresnel factor as well as the nonlinear susceptibility. This paper argues the relative importance of the two factors in general cases and provides a useful criterion to evaluate their importance. The effect of Fresnel dispersion becomes significant when the SFG spectrum involves a sharp and intense vibrational band as well as a large non-resonant background susceptibility, typically in some solid-liquid interfaces. A possible way to correct the effect of Fresnel dispersion is suggested using the heterodyne measurement.

SOMO-HOMO Conversion in Triplet Carbenes.

In this study, the SOMO-HOMO conversion has been shown for the first time in triplet carbenes embedded in cycloparaphenylene units. The high-lying HOMO originating from the curved π-conjugated system and the low-lying SOMO-1 originating due to the small carbene angle are the key to endowing this interesting electronic configuration. Furthermore, simple planar triplet carbenes such as fluorenylidene were found to possess SOMO-HOMO energy-converted electronic configurations.

Changes in mRNA of immune factors expressed by milk somatic cells of Holstein cows with hypocalcemia after calving.

Changes in immune factors expressed by milk somatic cells from Holstein cows with hypocalcemia after calving were investigated in this study. Fourteen multiparous Holstein cows after their 3rd or 4th calving in one farm were used. The cows were divided into 2 groups: 7 cows needing treatment due to onset of hypocalcemia (hypocalcemia group; age = 5.53 ± 0.27 years, parity = 3.14 ± 0.14) and 7 cows without health problems (control group; age = 5.88 ± 0.31 years, parity = 3.57 ± 0.26). Milk samples were collected aseptically using a cannula and mRNA of immune factors expressed by milk somatic cells were analyzed. Milk samples (50 mL) were collected from the right rear mammary gland of cows before milking at day 1 and weeks 1, 2, 4, and 8 after calving. All milk samples showed a negative reaction to the California Mastitis Test. Levels of relative interleukin (IL)-6 and cathelicidin in the hypocalcemia group were lower than those in the control group in weeks 1 to 8. A significant difference in relative IL-6 levels was found in week 4 (P < 0.05). These results suggest that levels of IL-6 expressed by milk somatic cells may be affected by hypocalcemia in dairy cows.

Dans la présente étude les modifications des facteurs immunitaires exprimées par les cellules somatiques du lait de vaches Holstein présentant une hypocalcémie après le vêlage ont été examinées. Quatorze vaches Holstein multipares après leur 3e ou 4e vêlage provenant d'une ferme ont été utilisées. Les vaches ont été réparties en deux groupes : sept vaches nécessitant un traitement en raison de l'apparition d'une hypocalcémie (groupe hypocalcémie; âge = 5,53 ± 0,27 ans, parité = 3,14 ± 0,14) et sept vaches sans problème de santé (groupe témoin; âge = 5,88 ± 0,31 ans, parité = 3,57 ± 0,26). Des échantillons de lait ont été prélevés de manière aseptique à l'aide d'une canule et l'ARNm des facteurs immunitaires exprimés par les cellules somatiques du lait a été analysé. Des échantillons de lait (50 mL) ont été prélevés dans la glande mammaire arrière droite des vaches avant la traite au jour 1 et aux semaines 1, 2, 4 et 8 après le vêlage. Tous les échantillons de lait ont montré une réaction négative au California Mastitis Test. Les niveaux relatifs d'interleukine (IL)-6 et de cathélicidine dans le groupe hypocalcémie étaient inférieurs à ceux du groupe témoin au cours des semaines 1 à 8. Une différence significative des taux relatifs d'IL-6 a été observée à la semaine 4 (P < 0,05). Ces résultats suggèrent que les taux d'IL-6 exprimés par les cellules somatiques du lait peuvent être affectés par l'hypocalcémie chez les vaches laitières.(Traduit par Docteur Serge Messier).

Influence of SOS-inducing agents on the expression of ArtAB toxin gene in Salmonella enterica and Salmonella bongori.

Salmonella enterica subspecies enterica serovar Typhimurium (S. Typhimurium) definitive phage type 104 (DT104), S. enterica subspecies enterica serovar Worthington (S. Worthington) and S. bongori produce ArtA and ArtB (ArtAB) toxin homologues, which catalyse ADP-ribosylation of pertussis toxin-sensitive G protein. ArtAB gene (artAB) is encoded on prophage in DT104 and its expression is induced by mitomycin C (MTC) and hydrogen peroxide (H2O2) that trigger the bacterial SOS response. Although the genetic regulatory mechanism associated with artAB expression is not characterized, it is thought to be associated with prophage induction, which occurs when the RecA-mediated SOS response is triggered. Here we show that subinhibitory concentration of quinolone antibiotics that are SOS-inducing agents, also induce ArtAB production in these Salmonella strains. Both MTC and fluoroquinolone antibiotics such as enrofloxacin-induced artA and recA transcription and artAB-encoding prophage (ArtAB-prophage) in DT104 and S. Worthington. However, in S. bongori, which harbours artAB genes on incomplete prophage, artA transcription was induced by MTC and enrofloxacin, but prophage induction was not observed. Taken together, these results suggest that SOS response followed by induction of artAB transcription is essential for ArtAB production. H2O2-mediated induction of ArtAB prophage and efficient production of ArtAB was observed in DT104 but not in S. Worthington and S. bongori. Therefore, induction of artAB expression with H2O2 is strain-specific, and the mode of action of H2O2 as an SOS-inducing agent might be different from those of MTC and quinolone antibiotics.

Prevalence of sub-clinical mastitis and its association with milking practices in an intensive dairy production region of Uganda.

A cross-sectional study was conducted to investigate the risk factors for sub-clinical mastitis (SCM) in Mbarara District, an intensive dairy production region of Uganda where hand-milking is dominant. In 30 farms, herd-level milking practices and SCM prevalence were studied. The SCM prevalences were 68.6% (417/608, 95% confidence interval (CI): 64.9-72.2%) and 39.2% (946/2,411, 37.3-41.2%) at the cow- and quarter-levels, respectively. A preventive factor for SCM was cow calmness at the end of milking (OR: 0.20, 95%CI: 0.05-0.79, P=0.021); a risk factor was rough teat-end (OR: 1.75, 95%CI: 1.14-2.68, P=0.011). Good cow hygiene was negatively associated with environmental mastitis (P=0.002). Appropriate hand-milking practices that avoid teat damage are expected to reduce SCM in Uganda.

Primary intrahepatic squamous cell carcinoma in a sika deer.

A white nodule was detected in the liver of a wild female sika deer. The nodule was histologically diagnosed as squamous cell carcinoma (SCC), and it transitioned into a hyperplastic and chronically inflamed intrahepatic bile duct showing Fasciola infection. Therefore, the tumor was demonstrated to have originated from the biliary epithelium of the intrahepatic bile duct. Hyperplastic and chronic inflammatory changes of the biliary epithelium might have contributed the carcinogenesis of the present case, as proposed in human primary intrahepatic SCC cases. To the best of our knowledge, this is the first reported case of primary intrahepatic SCC in an animal.

Relationship Between mRNA of Immune Factors Expressed by Milk Somatic Cells and Bacteria Present in Healthy Lactating Holstein Cows.

INTRODUCTION: The characteristics of immune factors in somatic cells from lactating dairy cows and their association with commensal bacteria in normal milk have not been clarified. This study investigated the relationship between the pathogenic bacteria in milk and somatic cell immune factors in healthy lactating cows. MATERIAL AND METHODS: In total 44 healthy Holstein cows were studied on one farm. Milk samples were collected aseptically using a cannula and these samples were cultured for detection of bacteria and analysis of mRNA of immune factors expressed by somatic cells. Cows were divided into two groups based on the microbial status of their milk samples: 12 cows showed bacteria in cultures (positive group), and the other 32 cows did not (negative group). RESULTS: The mRNA levels of IL-6, lactotransferrin, and cathelicidin expressed by somatic cells after milking decreased significantly compared to those before milking in both groups (P < 0.05). There were significantly lower mRNA levels of IL-6 and cathelicidin in the positive group compared to those in the negative group before milking. CONCLUSION: These results suggest that mRNA levels of IL-6 and cathelicidin expressed by the somatic cells may be affected by the presence of bacteria in healthy lactating dairy cows.

[Next-generation Sequencing Analysis of Bacterial Flora in Bovine Prototheca Mastitic Milk].

Prototheca zopfii is an achlorophyllic algae that causes bovine mastitis, resulting in a reduction in milk production and the secretion of thin, watery milk with white flakes. The aim of the present study was to evaluate the bacterial flora in the udder environment in protothecal mastitis. We used metagenomic next-generation sequencing (NGS) analysis to identify 16S rRNA genes from bacterial flora present in milk samples from protothecal mastitic dairy cows.Seven clinical strains of P. zopfii genotype 2 were isolated from 7 milk samples from 7 cases (Holstein cow) of protothecal mastitis; another 9 milk samples were obtained from 9 normal Holstein cows. The samples were collected in 2017 from cows in one dairy located in the Kushiro region in Hokkaido, Japan, which had a history of protothecal mastitis infection.The NGS produced 10,000 to 15,000 sequences in each DNA sample. To facilitate comparison, we grouped the sequencing results according to the culture-based protothecal mastitis diagnosis; sequences derived from the milk samples obtained from healthy cows were grouped separately.Sequences classified as Streptococcus spp., Pseudomonas spp. and Sphingomonas spp., Caulobacter segnis, Macrococcus caseolyticus, Methylobacterium tarhaniae, and Sphingomonas leidyi were the main sequences detected in the groups of samples from cows characterized by culture as having protothecal mastitis. Notably, Calothrix desertica (a cyanobacterium) sequences showed higher prevalence in these samples.To our knowledge, this is the first study to report that C. desertica sequences, effectively absent in the samples derived from healthy cows, are detected at high prevalence in samples from protothecal mastitic animals.

Universality of the Phytophthora mating hormones and diversity of their production profile.

Sexual reproduction of the plant pest Phytophthora is regulated by two mating hormones α1 and α2, which are acyclic oxygenated diterpenes first isolated from P. nicotianae A1 and A2 mating types, respectively. A previous report suggested the universality of these factors within this genus. To confirm this concept, we investigated 80 strains (19 species) of Phytophthora and a related genus, not only for the responsiveness to mating hormones but also for their productivity. The results indicated that among the 55 heterothallic strains, 24 (44%) responded to a mating hormone and 40 (73%) produced one or both hormones. These findings demonstrate the interspecies universality of mating hormones within the genus Phytophthora. Hormone productivity was found to be highly diverse and dependent on the strains used. Although the A2 mating type has been regarded as the α2 producer, 19 (59%) of the 32 A2-type strains produced both the hormones and two A2-type strains exclusively produced α1 in high yields. These results indicate that hormone biosynthesis in Phytophthora is universal but highly diverse and complex, and varies with culture conditions, providing us valuable information for future studies on the mechanism of mating hormone biosynthesis of Phytophthora.

Highly chemoselective and versatile method for direct conversion of carboxylic acids to ketones utilizing zinc ate complexes.

Various carboxylic acids were directly transformed into the corresponding ketones by utilizing organozinc ate complexes, which provide high chemoselectivity without any overreaction to the undesired tertiary carbinol, owing to formation of a stable tetrahedral zincioketal intermediate. This method offers good overall atom/step/pot economy and operational simplicity.

Biogenic polyamines capture CO2 and accelerate extracellular bacterial CaCO3 formation.

Bacteria, including cyanobacteria, as well as some fungi, are known to deposit calcium carbonate (CaCO(3)) extracellularly in calcium-containing artificial medium. Despite extensive investigation, the mechanisms involved in extracellular formation of CaCO(3) by bacteria have remained unclear. The ability of synthetic amines to remove carbon dioxide (CO(2)) from natural gas led us to examine the role of biogenic polyamines in CaCO(3) deposition by bacteria. Here, we demonstrated that biogenic polyamines such as putrescine, spermidine, and spermine were able to react with atmospheric CO(2) and the resultant carbamate anion was characterized by using nuclear magnetic resonance (NMR) analysis. Biogenic polyamines accelerated the formation of CaCO(3), and we artificially synthesized the dumbbell-shaped calcites, which had the same form as observed with bacterial CaCO3 precipitates, under nonbacterial conditions by using polyamines. The reaction rate of calcification increased with temperature with an optimum of around 40 °C. Our observation suggests a novel scheme for CO(2) dissipation that could be a potential tool in reducing atmospheric CO(2) levels and, therefore, global warming.

Relationship Between Tightness of the Hip Joint and Elbow Pain in Adolescent Baseball Players.

BACKGROUND: Repetitive tensile stresses from valgus torque can induce elbow injury in adolescent baseball players. Insufficient hip range of motion (ROM) can change throwing mechanics, reducing the transfer of energy from the lower to the upper extremities. Thus, hip ROM limitations may force the upper extremities to bear the burden of a strong throw. Improper pitching mechanics caused by insufficient hip ROM are thought to increase valgus torque on the elbow when throwing, increasing the risk of elbow injury. PURPOSE: To investigate the relationship between elbow pain and hip ROM in adolescent baseball players. STUDY DESIGN: Cross-sectional study; Level of evidence, 3. METHODS: A total of 122 adolescent baseball players with a mean age of 12.0 years (range, 6-14 years) participated in this study. Elbow pain, hip flexion angle, and the internal rotation angles of the hip at 0° and 90° of flexion were assessed. Participants were divided into a pain group and a normal group based on the pain assessment, and each hip angle was compared between groups using Student t tests. P values <.05 were considered statistically significant. RESULTS: Thirty-one of 122 players had elbow pain. The hip flexion angle of the trail leg was 121.9° ± 12.3° for the normal group and 111.2° ± 11.3° for the pain group (P = .0001). The plant leg hip flexion angles were 122.0° ± 12.4° and 113.6° ± 11.3° (P = .0014) for the normal and pain groups, respectively. The internal rotation angle at 0° of hip flexion of the trail leg was 49.4° ± 12.6° and 45.6° ± 8.8° (not significant), and of the plant leg was 49.1° ± 12.5° and 48.7° ± 11.5° (not significant), for the normal and pain groups, respectively. The internal rotation of the trail leg at 90° of hip flexion was 46.9° ± 13.3° in the normal group and 36.1° ± 15.7° in the pain group (P = .0005). In the plant leg, the internal rotation angle at 90° of hip flexion was 46.9° ± 12.2° and 36.4° ± 18.1° for the normal and pain groups, respectively (P = .0013). CONCLUSION: Limitations to hip flexion and internal rotation at 90° of hip flexion were risk factors for elbow injury. Differences in internal rotation angles between 0° and 90° of hip flexion may be important criteria for identifying adolescent baseball players at risk of elbow pain.