Starmerella kisarazuensis f.a., sp. nov., a novel yeast isolated from Trifolium pratense flowers.

Three yeast isolates, NBRC 115909T, NBRC 115910 and NBRC 116270, were isolated from Trifolium pratense (red clover) flowers collected from Kisarazu, Chiba, Japan. Analysis of the sequences of the D1/D2 domains of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) regions revealed that these isolates represent a single novel species within the genus Starmerella. Also, no ascospore formation was observed. The yeast isolates were closely related to Starmerella vitae UWOPS 00-107.2T and Starmerella bombi NRRL Y-17081T. They differed from S. vitae, the most closely related species with a validly published name, by ten nucleotide substitutions with two gaps in the D1/D2 domains and 20 nucleotide substitutions in the ITS region. Moreover, the three isolates exhibited distinct phenotypic characteristics from the closely related species. Therefore, we suggest that these three isolates represent a novel species, designated as Starmerella kisarazuensis f.a., sp. nov. The holotype is NBRC 115909T (isotype: CBS 18485T).

Role of DLA-DRB1 amino acids outside the shared epitope in dachshund susceptibility to immune-mediated polyarthritis.

Canine immune-mediated polyarthritis (IMPA) is an idiopathic disorder encompassing both erosive and non-erosive forms of rheumatoid arthritis (RA), with a clinical picture similar to human RA. Resemblance in major histocompatibility complex (MHC)-associated risk between the two was first noted within the specific amino acid motif known as the shared epitope (SE) on human leukocyte antigen DRB1. Following further identification of amino acids conferring risk for human RA outside the SE, this study was designed to examine amino acids both within and outside the classic SE in dachshunds, a breed with reported susceptibility to IMPA in Japan. Genome-wide association studies have linked positions 11, 13 and 71 with strong risk for human RA and important roles in antigen presentation to T cells. Sequence based genotyping of 16 case and 64 control dachshunds revealed strong associations comparable to human RA between IMPA risk and valine at position 11 (Val-11), phenylalanine at 13 (Phe-13), and arginine at 71 (Arg-71) on the dog leukocyte antigen (DLA)-DRB1 molecule (OR 2.89, 95%CI 1.3-6.4, p = 0.009), while association with the classic SE was significant only regarding homozygote frequency of the QRRAA haplotype-also carrying Val 11 and Phe 13 outside the SE (p = 0.04). Moreover, limited range in possible combinations of amino acids at positions 11, 13 and 71 starting with Val-11 among all DLA-DRB1 alleles registered with the GenBank and IPD-MHC canine databases, suggested potential of further single-breed analyses in dachshunds to clarify the disorder in terms of diagnosis, treatment, and epigenetic control, while clinical and immunopathogenetic similarities between human and dachshund RA also suggested the possibility of gaining insight into RA per se through study of canine IMPA as a spontaneous model of human RA.

North-to-South diversity of lipomycetaceous yeasts in soils evaluated with a cultivation-based approach from 11 locations in Japan.

To understand the species distribution, diversity, and density of lipomycetaceous yeasts in soil based on their north-to-south location in Japan, 1146 strains were isolated from soil samples at 11 locations from Hokkaido to Okinawa Prefecture and taxonomically characterized. Lipomycetaceous yeast strains were isolated efficiently from soil by selecting watery mucoid-like colonies on agar plates with nitrogen-depleted medium. Twenty-four (80%) of the 30 known species of the genus Lipomyces were isolated from the soil samples collected in Japan, including species recently proposed. Among the species isolated, L. starkeyi was the most predominant in Japan, except on Iriomote Island, Okinawa, and accounted for 60-98% of the isolated strains. Lipomyces yarrowii was the dominant species on Iriomote Island (64%). The second most dominant species were L. chichibuensis in Saitama Prefecture and L. doorenjongii from Yamaguchi to Okinawa Prefecture. The species diversity of lipomycetaceous yeasts was in Japan and the significant correlation with the latitude of the sampling sites was revealed.

Mn-Si-based nanoparticles-enhanced inhibitory effect on tumor growth and metastasis in photo-immunotherapy.

The anticancer effect of phototherapy has been limited by some factors, including the easy degradation of photo agents, the complex tumor microenvironment, and the limited immune activation capacity, which impedes its efficiency in inhibiting tumor growth and tumor metastasis. Herein, Mn-doped mesoporous silica nanoparticles were synthesized to load the photo agent of IR 780, which were further coated with Mn (IMM). Notably, the combination of IMM and an 808 nm laser irradiation simultaneously inhibited the growth of primary tumors and distant untreated tumors in a bilateral animal model, which could be attributed to the protection of IMM to IR 780, the regulation functions to the tumor microenvironment, as well as the enhanced immune activation capacity. This work highlighted an alternative strategy for enhancing the inhibitory effect on both tumor growth and tumor metastasis in the combinational anticancer therapy of phototherapy and immunotherapy (photo-immunotherapy).

Mid-infrared laser heterodyne spectrometer by hollow optical fiber and its newly designed coupler.

We demonstrate a newly designed, to the best of our knowledge, hollow optical fiber coupler for a mid-infrared (IR) laser heterodyne spectrometer that mixes a targeted light source with local oscillator (LO) light. The hollow fiber achieves a high transmission efficiency ∼80-90%/m, not only for a coherent laser source but also for an incoherent blackbody source. The branching characteristics of the hollow optical fiber coupler are found to be strongly dependent on the curvature and length of the input port fiber, indicating that the branching ratio could be designed independently for each input port. Our laboratory measurements demonstrate that the branching ratio and transmittance of the coupler can be varied by coupling a flexible fiber to the input side owing to the excitation of higher-order modes. Using the hollow optical fiber coupler, a high-resolution emission spectrum of the quantum cascade laser at 10.3 µm for our C O 2 laser-based heterodyne spectrometer is successfully achieved. Using a C O 2 laser with a hollow fiber and a blackbody as a direct input signal in free space, we obtain the sensitivity performance of IR laser heterodyne spectrometer as 2000-3000 K of the system noise temperature. This suggests that the transmission of a coherent LO laser through a hollow optical fiber has almost the same sensitivity for the IR heterodyne detection as that without a fiber.

Alternate oscillations of Martian hydrogen and oxygen upper atmospheres during a major dust storm.

Dust storms on Mars play a role in transporting water from its lower to upper atmosphere, seasonally enhancing hydrogen escape. However, it remains unclear how water is diurnally transported during a dust storm and how its elements, hydrogen and oxygen, are subsequently influenced in the upper atmosphere. Here, we use multi-spacecraft and space telescope observations obtained during a major dust storm in Mars Year 33 to show that hydrogen abundance in the upper atmosphere gradually increases because of water supply above an altitude of 60 km, while oxygen abundance temporarily decreases via water ice absorption, catalytic loss, or downward transportation. Additionally, atmospheric waves modulate dust and water transportations, causing alternate oscillations of hydrogen and oxygen abundances in the upper atmosphere. If dust- and wave-driven couplings of the Martian lower and upper atmospheres are common in dust storms, with increasing escape of hydrogen, oxygen will less efficiently escape from the upper atmosphere, leading to a more oxidized atmosphere. These findings provide insights regarding Mars' water loss history and its redox state, which are crucial for understanding the Martian habitable environment.

A pilot study of microRNA assessment as a means to identify novel biomarkers of spontaneous osteoarthritis in dogs.

MicroRNAs (miRNAs) are important regulators of intercellular signaling and are promising biomarkers in osteoarthritis (OA). In this study, comprehensive analysis was performed to identify miRNAs involved in the pathogenesis of spontaneous OA in dogs. Dogs diagnosed with OA based on radiography and arthroscopy of the stifle joint were included in the OA group. Dogs without any evidence of orthopedic disease were included in the unaffected group. To investigate miRNA expression levels, RNA sequencing analysis (RNA-seq) was performed in synovial tissue (OA group: n = 3, Unaffected group: n = 3) and RT-qPCR was performed in synovial tissue, synovial fluid and serum (OA group: n = 17, Unaffected group: n = 6), and compared between the two groups. The RNA-seq results showed that 57 miRNAs were significantly upregulated and 42 were significantly downregulated in the OA group. Specifically, miR-542 and miR-543 expression levels in the synovial tissue, synovial fluid, and serum were consistently higher in the OA group than in the unaffected group, suggesting that these miRNAs may be used as biomarkers for detecting canine OA. This is the first report to comprehensively analyze the expression patterns of miRNAs in the synovial tissue of dogs with spontaneous OA.

Tumor microenvironment regulation - enhanced radio - immunotherapy.

Radiotherapy (RT) is frequently utilized for cancer treatment in clinical practice and has been proved to have immune stimulation potency in recent years. However, its inhibitory effect on tumor growth, especially on tumor metastasis, is still limited by many factors, including the complex tumor microenvironment (TME). Therefore, the TME - regulating SiO2@MnO2 nanoparticles (SM NPs) were prepared and applied to the combination of RT and immunotherapy. In a bilateral animal model, SM NPs not only enhanced the inhibitory effect of RT on primary tumor growth, but also strengthened the abscopal effect to inhibit the growth of distant untreated tumors. As for the distant untreated tumor, 40% of mice showed complete inhibition of tumor growth and 40% showed a suppressed tumor growth. Moreover, SM NPs showed modulation functions for TME through inducing the increase in intracellular levels of oxygen and reactive oxygen species after their reaction with hydrogen peroxide and the main antioxidative agent glutathione in TME. Lastly, SM NPs also effectively induced the increase in the amounts of cytokines secreted by macrophage - like cells, indicating modulation functions for immune responses. This work highlighted a potential strategy of simultaneously inhibiting tumor growth and metastasis through the regulation of TME and immune responses by SM NPs - enhanced radio - immunotherapy.

Step-by-step protocols for non-viral derivation of transgene-free induced pluripotent stem cells from somatic fibroblasts of multiple mammalian species.

Potentials of immortal proliferation and unlimited differentiation into all the three germ layers and germ cells in induced pluripotent stem cells (iPSCs) render them important bioresources for in vitro reconstitution and modeling of intravital tissues and organs in various animal models, thus contributing to the elucidation of pathomechanisms, drug discovery and stem cell-based regenerative medicine. We previously reported promising approaches for deriving transgene-free iPSCs from somatic fibroblasts of multiple mammalian species by episomal vector or RNA transfection, although the respective step-by-step protocols and the combinatorial usage of these methods, which achieved high induction efficiency, have not been described in the literature so far. Here, we provide a detailed step-by-step description of these methods with critical tips and slight modifications (improvements) to previously reported methods. We also report a novel method for the establishment of iPSCs from the Syrian hamster (also known as golden hamster; Mesocricetus auratus), a unique animal model of hibernation. We anticipate this methodology will contribute to stem cell biology and regenerative medicine research.

Tumor microenvironment-regulated nanoplatforms for the inhibition of tumor growth and metastasis in chemo-immunotherapy.

Chemotherapy is one of the major clinical anticancer therapies. However, its efficiency is limited by many factors, including the complex tumor microenvironment (TME). Herein, manganese-doped mesoporous silica nanoparticles (MM NPs) were constructed and applied to regulate the TME and enhance the efficiency of the combination of chemotherapy and immunotherapy (chemo-immunotherapy). Notably, the combination of MM NPs, doxorubicin hydrochloride, and immune checkpoint inhibitors enhanced the synergistic efficiency of chemo-immunotherapy in a bilateral animal model, which simultaneously inhibited the growth of primary tumors and distant untreated tumors. Moreover, Mn-doping endowed MSNs with six new regulatory functions for the TME by inducing glutathione depletion, ROS generation, oxygenation, cell-killing effect, immune activation, and degradation promotion. These results demonstrated that MM NPs with TME regulatory functions can potentially improve the efficiency of chemo-immunotherapy.

Evaluation of Hindlimb Deformity and Posture in Dogs with Grade 2 Medial Patellar Luxation during Awake Computed Tomography Imaging while Standing.

OBJECTIVE: The aim of this study was to determine the degree of bone deformities and hindlimb postural abnormalities in a standing position in awake Toy poodles with and without grade 2 medial patellar luxation (MPL) using high speed 320-row computed tomography (CT). METHODS: The limbs with grade 2 MPL (MPL-G2 group) and without any orthopaedic disorders (control group) were imaged in a standing position, without sedation or anaesthesia, using CT. In MPL-G2 group, images were obtained when the patella was luxated (G2-L group) and reduced (non-luxation, G2-NL group). Bone morphologies of the femur and tibia were quantified three-dimensionally. Hindlimb standing posture was evaluated by measuring femoral rotation and abduction angles, tibial rotation angle, metatarsal rotation angle, foot rotation angle, angle between the femoral anatomical axis and the mechanical axis of hindlimb and stifle joint line convergence angle. RESULTS: There were no significant differences in bone morphologic parameters between the MPL-G2 group (5 limbs) and the control group (6 limbs). In the G2-NL group, there were no significant hindlimb postural abnormalities. In contrast, in the G2-L group, significant hindlimb postural abnormalities including external rotation of femur, internal rotation of tibia and foot, external rotation of tarsal joint, large stifle joint convergence angle, genu varum and toe-in standing were observed. CONCLUSION: Dogs with grade 2 MPL have no bone deformities but show abnormal standing posture when the patella is luxated.

The nightside cloud-top circulation of the atmosphere of Venus.

Although Venus is a terrestrial planet similar to Earth, its atmospheric circulation is much different and poorly characterized1. Winds at the cloud top have been measured predominantly on the dayside. Prominent poleward drifts have been observed with dayside cloud tracking and interpreted to be caused by thermal tides and a Hadley circulation2-4; however, the lack of nightside measurements over broad latitudes has prevented the unambiguous characterization of these components. Here we obtain cloud-tracked winds at all local times using thermal infrared images taken by the Venus orbiter Akatsuki, which is sensitive to an altitude of about 65 kilometres5. Prominent equatorward flows are found on the nightside, resulting in null meridional velocities when these are zonally averaged. The velocity structure of the thermal tides was determined without the influence of the Hadley circulation. The semidiurnal tide was found to have an amplitude large enough to contribute to the maintenance of the atmospheric superrotation. The weakness of the mean meridional flow at the cloud top implies that the poleward branch of the Hadley circulation exists above the cloud top and that the equatorward branch exists in the clouds. Our results should shed light on atmospheric superrotation in other celestial bodies.

Analysis of the light regulatory mechanism in carotenoid production in Rhodosporidium toruloides NBRC 10032.

Light stimulates carotenoid production in an oleaginous yeast Rhodosporidium toruloides NBRC 10032 by promoting carotenoid biosynthesis genes. These genes undergo two-step transcriptional activation. The potential light regulator, Cryptochrome DASH (CRY1), has been suggested to contribute to this mechanism. In this study, based on KU70 (a component of nonhomologous end joining (NHEJ)) disrupting background, CRY1 disruptant was constructed to clarify CRY1 function. From analysis of CRY1 disruptant, it was suggested that CRY1 has the activation role of the carotenogenic gene expression. To obtain further insights into the light response, mutants varying carotenoid production were generated. Through analysis of mutants, the existence of the control two-step gene activation was proposed. In addition, our data analysis showed the strong possibility that R. toruloides NBRC 10032 is a homo-diploid strain.

Functional Anatomy of the Craniolateral and Caudomedial Bundles of the Caudal Cruciate Ligament in Beagle Dogs.

OBJECTIVE: The aim of this study was to investigate functional anatomy of the craniolateral bundle (CrLB) and caudomedial bundle (CdMB) of caudal cruciate ligament in normal Beagles. MATERIALS AND METHODS: Twelve stifle joints of 12 Beagle cadavers that were euthanatized for reasons other than musculoskeletal disease were used. Anatomical characteristics of femoral and tibial attachment regions of the caudal cruciate ligament were described, and location and area of the footprints of the CrLB and CdMB were objectively determined. The CrLB was artificially divided into two equal fibre bundle units and the CdMB was artificially divided into four equal bundle units. Changes in the fibre arrangement were recorded during stifle range of motion, and the tension of each fibre bundle unit at various stifle joint angles was objectively measured. RESULTS: The CrLB was attached to the intercondylar fossa of the femur, and the CdMB was attached to the medial surface of the medial condyle of the femur. The centre of the CrLB attachment site was located craniomedial to that of the CdMB on the tibia. During stifle range of motion, each divided fibre bundle unit in the CrLB and CdMB was twisted externally. The tension of entire CdMB tended to be higher than that of entire CrLB at 180 to 135 degrees of the stifle angles. In full extension angle, the tension of entire CdMB was 1.55 times higher than that of entire CrLB. CLINICAL SIGNIFICANCE: This study contributes to understanding of the biomechanics and the function of the caudal cruciate ligament, which help to elucidate the pathophysiology of caudal cruciate ligament tear and to develop stifle joint stabilization surgery.

Variations in gene expression levels with severity of synovitis in dogs with naturally occurring stifle osteoarthritis.

Osteoarthritis (OA) is one of the major causes of chronic pain in dogs. However, the pathogenesis of OA has not been fully understood in dogs. The objective of this study was to comprehensively investigate the mRNA expression levels of proinflammatory cytokines, inflammatory mediators, nerve growth factor and its receptor, and matrix metalloproteinases in the synovium of dogs with spontaneous OA as well as to elucidate their relationships with the severity of synovitis. Dogs that were diagnosed with stifle OA on the basis of radiographic findings were included, and the degree of synovitis was observed using stifle arthroscopy. The dogs were assigned to two different groups depending on their synovitis scores: the low-grade group (score of 1 or 2; n = 8) and high-grade group (score of 3 to 5; n = 18). The dogs showing no evidence of orthopedic disease were included in the control group (n = 6). Synovial tissue samples were collected from the sites at which synovitis scores were assessed using arthroscopy. Total RNA was extracted from the collected synovial tissue, and cDNA was synthesized. Subsequently, RT-qPCR were performed using canine-specific primer sets for IL1B, IL6, CXCL8, TNF, TGFB1, PTGS2, PTGES, MMP3, MMP13, NGF, NTRK1, and PTGER4. Expression levels of IL1B, IL6, CXCL8, and MMP13 were significantly higher in the high-grade group than in the control group. In addition, expression levels of IL1B, CXCL8, TNF, and PTGS2 were significantly higher in the high-grade group than in the low-grade group. Expression levels of IL1B, IL6, CXCL8, TNF, PTGS2, and PTGER4 showed significant positive correlation with synovitis score. In conclusion, all mRNA expression levels in the synovial membrane varied according to the degree of synovitis in dogs with spontaneous OA. Thus, this study may partially elucidate the pathogenesis of synovitis in dogs with spontaneous OA.

Treatment of methicillin-resistant Staphylococcus aureus infection following tibial plateau leveling osteotomy in a dog.

BACKGROUND: In recent years, surgical site infections caused by drug-resistant pathogens have emerged as a cause of concern in small animal practice. In this report, methicillin-resistant Staphylococcus aureus (MRSA) infections associated with tibial plateau leveling osteotomy (TPLO) is reported. However, there have been no reports on the treatment of MRSA infection following TPLO in dogs. This case report describes the use of a combination of vancomycin and rifampicin to treat MRSA infection following TPLO in a dog. CASE DESCRIPTION: A 7-year-old spayed female American cocker spaniel was referred for right hind limb lameness that did not improve with conservative treatment. The dog was diagnosed with cranial cruciate ligament rupture, for which TPLO was performed. Once the surgical wound was closed, the dog licked the skin on the surgical site, causing the injury to dehisce. MRSA was detected from the purulent discharge, and chloramphenicol was then administered based on the drug sensitivity test results. Because of the continued drainage, the implants were removed after the bone union of the osteotomy site was observed. Since this did not provide any relief to the existing condition, the antibiotic was changed to vancomycin at 132 days after TPLO surgery, and the infected location was cleaned many times through a drain tube placed into the tibia. However, the infection could not be controlled. Thus, a rifampicin and vancomycin combination was started. As a result, the purulent discharge disappeared and the fistula entirely closed on the 154th day after TPLO surgery. CONCLUSION: A combination of rifampicin and vancomycin may be effective for treating MRSA infection at the surgical site following TPLO surgery that does not heal despite implant removal and administration of vancomycin.

First experimental verification of the neutron field of Nagoya University Accelerator-driven neutron source for boron neutron capture therapy.

As the commissioning phase of the Nagoya University Accelerator-driven Neutron Source for boron neutron capture therapy, in-phantom thermal neutron flux measurements were conducted using a small 6LiF/Eu:CaF2 scintillator detector and activation foils. The spatial distribution of the measured thermal neutron flux agreed with the Monte Carlo simulation results. Based on this comparison, the free-in-air neutron spectrum, calculated at the exit aperture, was verified and the epithermal neutron flux, at a 0.25 mA proton current, was evaluated as (1.49 ± 0.10) × 107 n/(cm2 s).

Design for stray-light reduction to a Martian ionospheric imager.

We demonstrate a new design high-contrast optics suitable for use on a stray-light reduction. Two-dimensional distribution of diffracted light was numerically analyzed and confirmed by a laboratory measurement to determine its characteristics. Laboratory measurements confirm stray-light performance with ∼10-8 rejection viewing as close as 1.0° to a bright-light source. The present improvement replaces the conventional normal-edge vane by an edge with microscopic Gaussian-shaped structures. This permits simplified fabrication, provides even more weight reduction, and delivers improved performance for a given size.

Utility of a novel activity monitor assessing physical activities and sleep quality in cats.

Osteoarthritis in cats is more prevalent because cats are living longer with advancement in veterinary medicine. Objective evaluation of behavioral changes in cats with osteoarthritis can facilitate an early diagnosis. The objective of this study was to investigate the utility of a novel activity monitor for analyzing physical activities and sleep quality in cats. First, a novel activity monitor (Plus Cycle®; JARMeC, Kanagawa, Japan), with a built-in three-directional accelerometer and an air pressure sensor specifically designed for cats, was compared to a human activity monitor (Actical®; Philips Respironics, OR, USA) previously used in cats (n = 10). Second, the validity of the measurement accuracy of the amount of physical activity, the number of vibrations, the number of jumps, and the resting and sleeping time was evaluated using Plus Cycle® in healthy cats (n = 6). Finally, the effects of gender and age of cats and time of day on the amount of physical activity, the number of vibrations, the number of jumps, and the resting and sleeping time were investigated in client-owned cats (n = 61). There were strong correlations between Plus Cycle® and Actical® in total activity (p < 0.05) and activity intensity (p < 0.05). When the physical activities were measured using Plus Cycle® in healthy cats, those data were quantified with high accuracy. In addition, it was also found to be very accurate in discriminating the resting and sleeping conditions of cats. In client-owned cats, there were no significant differences with respect to gender in any measured traits. The amount of physical activity and the number of jumps significantly decreased with the age of the cat. In contrast, the resting and sleeping times significantly increased with the age of the cat. In conclusion, Plus Cycle® can accurately and objectively assess physical activities and sleep quality with age of the cat, suggesting that this novel activity monitor can be used to manage the feline musculoskeletal health.

How waves and turbulence maintain the super-rotation of Venus' atmosphere.

Venus has a thick atmosphere that rotates 60 times as fast as the surface, a phenomenon known as super-rotation. We use data obtained from the orbiting Akatsuki spacecraft to investigate how the super-rotation is maintained in the cloud layer, where the rotation speed is highest. A thermally induced latitudinal-vertical circulation acts to homogenize the distribution of the angular momentum around the rotational axis. Maintaining the super-rotation requires this to be counteracted by atmospheric waves and turbulence. Among those effects, thermal tides transport the angular momentum, which maintains the rotation peak, near the cloud top at low latitudes. Other planetary-scale waves and large-scale turbulence act in the opposite direction. We suggest that hydrodynamic instabilities adjust the angular-momentum distribution at mid-latitudes.