A rumen virosphere with implications of contribution to fermentation and methane production, and endemism in cattle breeds and individuals.

Viruses have a potential to modify the ruminal digestion via infection and cell lysis of prokaryotes, suggesting that viruses are related to animal performance and methane production. This study aimed to elucidate the genome-based diversity of rumen viral communities and the differences in virus structure between individuals and cattle breeds and to understand how viruses influence on the rumen. To these ends, a metagenomic sequencing of virus-like particles in the rumen of 22 Japanese cattle, including Japanese Black (JB, n = 8), Japanese Shorthorn (n = 2), and Japanese Black sires × Holstein dams crossbred steers (F1, n = 12) was conducted. Additionally, the rumen viromes of six JB and six F1 that were fed identical diets and kept in a single barn were compared. A total of 8,232 non-redundant viral genomes (≥5-kb length and ≥50% completeness), including 982 complete genomes, were constructed, and rumen virome exhibited lysogenic signatures. Furthermore, putative hosts of 1,223 viral genomes were predicted using tRNA and clustered regularly interspaced short palindromic repeat (CRISPR)-spacer matching. The genomes included 1 and 10 putative novel complete genomes associated with Fibrobacter and Ruminococcus, respectively, which are the main rumen cellulose-degrading bacteria. Additionally, the hosts of 22 viral genomes, including 2 complete genomes, were predicted as methanogens, such as Methanobrevibacter and Methanomethylophilus. Most rumen viruses were highly rumen and individual specific and related to rumen-specific prokaryotes. Furthermore, the rumen viral community structure was significantly different between JB and F1 steers, indicating that cattle breed is one of the factors influencing the rumen virome composition.IMPORTANCEHere, we investigated the individual and breed differences of the rumen viral community in Japanese cattle. In the process, we reconstructed putative novel complete viral genomes related to rumen fiber-degrading bacteria and methanogen. The finding strongly suggests that rumen viruses contribute to cellulose and hemicellulose digestion and methanogenesis. Notably, this study also found that rumen viruses are highly rumen and individual specific, suggesting that rumen viruses may not be transmitted through environmental exposure. More importantly, we revealed differences of viral communities between JB and F1 cattle, indicating that cattle breed is a factor that influences the establishment of rumen virome. These results suggest the possibility of rumen virus transmission from mother to offspring and its potential to influence beef production traits. These rumen viral genomes and findings provide new insights into the characterizations of the rumen viruses.

Symptoms and health-related quality of life in patients with newly diagnosed multiple myeloma: a multicenter prospective cohort study.

OBJECTIVE: Existing cross-sectional observational studies indicate that patients with multiple myeloma experience negative physical and psychological symptoms and low health-related quality of life. The study aim was to determine symptom prevalence, health-related quality of life and symptoms associated with health-related quality of life in patients with newly diagnosed multiple myeloma. METHODS: This multicenter longitudinal cohort study was conducted in four hospitals in Japan. Patients with newly diagnosed multiple myeloma were asked to report their symptom intensity and health-related quality of life using validated questionnaires at three points: at diagnosis (T1), 1 month (T2) and 12 months after diagnosis (T3). Symptoms associated with health-related quality of life were explored using a mixed-effects model. RESULTS: A total of 106 patients completed the assessment at T1. The symptoms more than 30% of patients reported were pain, disturbed sleep and distress at T1, pain, dry mouth, disturbed sleep and fatigue at T2, fatigue, numbness of tingling and pain and numbness or tingling at T3. Pain and depression were significantly associated with health-related quality of life negatively. CONCLUSIONS: The finding suggests that more than 30% of multiple myeloma patients suffered from pain and various symptoms and they received suboptimal palliative care within a year after starting initial chemotherapy. Pain and depression should be the main targets of interventions to improve health-related quality of life in this population.

Mode-selective inhibitory effects of eugenol on the mouse TRPV1 channel.

The transient receptor potential vanilloid 1 (TRPV1) channel is a polymodal receptor in sensory nerves and involved in pain sensation. TRPV1 has at least three distinct activation modes that are selectively induced by different stimuli capsaicin, noxious heat, and protons. Although many mode-selective TRPV1 antagonists have been developed for their anticipated analgesic effects, there have been few successful reports because of adverse effects due to burn injuries and hyperthermia. Eugenol is a vanilloid that has been used as an analgesic in the dental treatment, and its TRPV1 activation ability has been reported. However, our knowledge about the underlying mechanisms of the antagonistic effects of eugenol on TRPV1 activation induced by three different modes is limited. Here, we show that eugenol dose-dependently inhibited the capsaicin-activated inward currents of mouse TRPV1 expressed in human embryonic kidney 293 (HEK293) cells. Under low pH conditions, low concentrations of eugenol only enhanced the proton-induced TRPV1 currents, whereas high eugenol concentrations initially potentiated but then immediately abrogated TRPV1 currents. Finally, eugenol had no modulatory effects on heat-activated TRPV1 in electrophysiological and Fura-2-based Ca2+ imaging experiments. Our results demonstrate that eugenol is a mode-selective antagonist of TRPV1 and can be evaluated as a lead compound of analgesics targeting TRPV1 without serious side effects.

Current trends in use of intracanal medications in dental care facilities: questionnaire-based survey on training dental hygienists at educational institutions.

The success of root canal therapy is dependent not only on removal of infected pulp (pulpectomy) followed by root canal enlargement, but also on the pharmacological effects of intracanal medications. Various intracanal medications are used. Formaldehyde preparations such as formocresol were common in the past, but these are no longer used in Europe or the US due to the biological toxicity of formaldehyde. In this study, a questionnaire was used to determine current trends in the use of intracanal medications at dental care facilities where dental hygiene students undergo practical training. The questionnaire comprised questions regarding the types of frequently used intracanal medications and their methods of application at dental care facilities in Saitama and Shizuoka prefectures. The results indicated that calcium hydroxide preparations were more commonly used in Europe or the US. However, these results also revealed that formaldehyde preparations were frequently used, which slightly differs from the scenario in Europe and the US. This study revealed that multiple intracanal medications were used for root canal therapy. Furthermore, it was also observed that cotton plugs were generally used as applicator tips for intracanal medications, whereas the use of absorbent paper points was relatively uncommon. The results suggest that the cost of absorbent paper points needs to be reduced.

Identification of 146 Metagenome-assembled Genomes from the Rumen Microbiome of Cattle in Japan.

The rumen contains a complex microbial ecosystem that degrades plant materials, such as cellulose and hemicellulose. We herein reconstructed 146 nonredundant, rumen-specific metagenome-assembled genomes (MAGs), with ≥50% completeness and <10% contamination, from cattle in Japan. The majority of MAGs were potentially novel strains, encoding various enzymes related to plant biomass degradation and volatile fatty acid production. The MAGs identified in the present study may be valuable resources to enhance the resolution of future taxonomical and functional studies based on metagenomes and metatranscriptomes.

Role of a long-chain alkyl group in sulfated alkyl oligosaccharides with high anti-HIV activity revealed by SPR and DLS.

Although a long-chain alkyl group in sulfated oligosaccharides can enhance the anti-HIV activity, the exact mechanism is unclear. To elucidate the role of the long-chain alkyl group, its interaction with a liposome (100 nm) as a HIV model was investigated by surface plasmon resonance and dynamic light scattering. The newly synthesized sulfated 1-(decadecyl-1, 2, 3-triazole)-1-deoxy- maltoheptaoside bearing the long-chain alkyl group was found to interact with the liposome. The particle size increased and the ζ potential was negative, indicating that the sulfated alkyl maltoheptaoside was attached to the liposome by the long-chain alkyl group and the fixed sulfated maltoheptaoside moiety was covered on the liposome. Thus, the long-chain alkyl group penetrates and is fixed into the lipid bilayer of HIV and the sulfated maltoheptaose moiety with negatively charged sulfate groups was electrostatically interacted with HIV gp120 molecule with positively charged amino acids to achieve the inhibition of HIV infection.

Interaction between sulfated 3-O-octadecyl-α-(1→6)-d-glucan and liposomes analyzed by surface plasmon resonance.

To elucidate the role of long alkyl group in sulfated poly- and oligosaccharides on anti-HIV activity, the interaction between sulfated 3-O-octadecyl-(1→6)-α-d-glucopyranan with potent anti-HIV activity and liposomes with diameters of 58 ± 20 nm and 107 ± 28 nm as models of HIV were investigated. SPR measurements of sulfated 3-O-octadecyl-(1→6)-α-d-glucopyranans bearing 2.8 mol% of the octadecyl group and the liposome (diameter = 58 ± 20.0 nm and ζ=0 mV) resulted in an apparent association- ka = 6 × 105 1/M, a dissociation-rate kd = 4 × 10-4 1/s, and a dissociation constants KD = 8 × 10-10 M. The particle size of the sulfated 3-O-octadecyl-(1→6)-α-d-glucopyranan (67 ± 14 nm) measured by DLS increased to 104 ± 25 nm, whereas the ζ potential (-29 mV) was unchanged (-33 mV). For dextran sulfate without an alkyl group, no interaction was observed. These results suggest that the long octadecyl group penetrated into the liposome and sulfated glucopyranan was covered on the liposome to increase the anti-HIV activity. The 107 nm liposome exhibited similar results.

Pentameric ellagitannin oligomers in melastomataceous plants--chemotaxonomic significance.

The pantropical plant family Melastomataceae produces characteristic hydrolyzable tannin oligomers. The latter in this family are distinguished from those in other plant families by the fact that the oligomers from dimers to tetramers are composed of two different alternating monomeric units: casuarictin and pterocaryanin C. These oligomers are metabolites that are produced by intermolecular C-O oxidative coupling between the monomers (or their desgalloyl-or des-hexahydroxydiphenoyl derivatives) forming a valoneoyl group as the link between monomers. The chemotaxonomically significant oligomerization pattern of melastomataceous plants provided helpful suggestions for determining the structures of new oligomers (nobotanins Q-T and melastoflorins A-D) isolated from Monochaetum multiflorum, which belongs to this family. Melastoflorins A-D were characterized as pentamers, which are the largest hydrolyzable tannins composed of different monomeric units.

Proanthocyanidin glycosides and related polyphenols from cacao liquor and their antioxidant effects.

Purification of polar fractions from cacao liquor extracts gave 17 phenolics including four new compounds. The new compounds were characterized as a C-glycosidic flavan, an O-glycoside of a dimeric and two O-glycosides of trimeric A-linked proanthocyanidins, on the basis of spectroscopic data. Isolated polyphenols showed inhibitory effects on nicotinamide adenine dinucleotide phosphate-dependent lipid peroxidation in microsomes and on the autoxidation of linoleic acid. These effects were attributed to the radical-scavenging activity in the peroxidation chain reactions, based on the findings that the cacao polyphenols effectively scavenged the 1,1-diphenyl-2-picrylhydrazyl radical.

Methotrexate-related Epstein-Barr virus-associated lymphoproliferative disorder occurring in the gingiva of a patient with rheumatoid arthritis.

It is well recognized that patients with immunodeficiency have a high risk of development of lymphoproliferative disorders (LPDs), and Epstein-Barr virus (EBV) is associated with the occurrence of LPDs. Methotrexate (MTX) is one of the common cause of iatrogenic-associated LPD, and approximately 40-50% of MTX-related LPD cases occur in extranodal sites. However, the occurrence of MTX-related LPD in the gingiva is extremely rare. Herein, we report the fourth documented case of MTX-related EBV-associated LPD occurring in the gingiva of a patient with rheumatoid arthritis (RA). A 76-year-old Japanese female with a 10-year history of RA, who was treated with MTX and infliximab, presented with a tumorous lesion in the gingiva. Biopsy of the gingiva tumor revealed diffuse proliferation of large-sized lymphoid cells with cleaved nuclei containing conspicuous nucleoli. These lymphoid cells were CD20- and EBER-positive. Therefore, a diagnosis of MTX-related EBV-associated LPD showing features of diffuse large B-cell lymphoma (DLBCL) that occurred in the gingiva was made. Although the occurrence of LPD in the oral region, as seen in the present case, is rare, the prevalence of this disorder may be on the rise due to the increased number of patients undergoing immunosuppression therapy. Moreover, immunosenescence can also be a cause of EBV-associated LPD. Therefore, recognition of the occurrence of this disorder in the oral cavity and consideration of the clinical history can facilitate the correct diagnosis.

Synthesis of oligosaccharide-branched ribofuranan by ring-opening polymerization of a new anhydroribo trisaccharide monomer.

A new anhydroribotrisaccharide monomer, A2B3LR (1), was synthesized and ROP was carried out to elucidate the polymerizability and to obtain oligosaccharide-branched polysaccharides with defined structures. The new trisaccharide monomer was found to be polymerized readily with BF(3) . OEt(2) as a catalyst at -40 degrees C to give a lactose-branched polymer. Copolymerization with ADBR gave the corresponding copolymers in good yields. After removal of protective benzyl groups, D-lactose-branched ribofuranans with free hydroxyl groups were obtained in good yields. The structure of polymers was analyzed by (1)H, 13C, and two-dimensional NMR measurements, suggesting that D-lactose-branched ribofuranans had (1 --> 5)-alpha stereoregularity.

Enzymatic synthesis of polyphenols from highly phenolic lignin-based polymers (lignophenols).

Peroxidase-catalyzed polymerization of lignin-based macromonomers (lignophenols), lignocatechol and lignocresol, prepared by phenolation of lignin with catechol or p-cresol, was carried out in aqueous organic solvent mixtures. The two lignophenols were polymerized to give cross-linked polymers. The highest yield of polymerization (83%, w/w) was obtained with lignocatechol, and the maximum yield for the polymerization of lignocresol was 55% (w/w). Pyrolysis GC-MS analysis of polymers indicated that the polymerization of lignophenols involved the oxidative coupling of the introduced phenol derivatives.