Glutamate in the medium of Lactiplantibacillus plantarum FL-664 affects the production of IL-12(p40) on murine spleen cells.

Lactic acid bacteria (LAB) have been attracting attention for their effects on innate immunity, and therefore, it is required to develop an efficient culturing method while maintaining their functionality. In this study, first, we compared the growth and functionality of LAB cultured on food grade (FG) medium with those on standard LAB medium and found that LAB cultured in the FG medium were smaller in cell size with high yield and had a higher ability to induce IL-12(p40) production by murine spleen cells in vitro. Moreover, the higher the glutamate concentration in the medium, the smaller the cell size, and the higher the yield and the higher the ability to induce IL-12 production. Addition of glutamate to the culture medium changes the size of LAB and affects their ability to induce IL-12(p40) production. In conclusion, regulating the concentration of glutamate would be important in the efficient culturing of functional LAB.

Airway Compromise in Mandibular Fracture.

ABSTRACT: Airway compromise is life threatening complication in the maxillofacial fracture. However, the incident of the airway compromise following mandibular fracture was little known. Maxillofacial fractures during the past 3 years from 2017 to 2020 were investigated retrospectively. There were 154 patients with maxillofacial fractures were noted in the past 3 years. Mandibular fractures accounted for 93% of the maxillofacial fracture. The most common cause of injury was falls (75%), followed by sports (12%). Of these, 3 cases (1.9%) needed an airway management at the time of initial treatment, and all were mandibular fractures. The average age was 55.7 ±â€Š10 years and all were male. The cause of the injury was high-energy trauma including traffic accident and work-related injury. Endotracheal intubation was performed at the time of initial treatment. All cases were communicated mandibular fracture included symphysis and para-symphysis region, and the bone fragments including the genial tubercle were largely displaced inward.

Sulfonamide antibiotics inhibit RNAi by binding to human Argonaute protein 2 PAZ.

We found that sulfisomidine, a sulfonamide antibiotic, potently binds to the Piwi/Argonaute/Zwille (PAZ) domain of human Argonaute protein 2 and inhibits RNA interference (RNAi). To elucidate the effect on RNAi of strong affinity of the 3'-ends in small interfering RNA (siRNA) to the PAZ domain, chemically modified siRNAs bearing sulfisomidine at the 3'-end were synthesized.

Adjuvant Activity Enhanced by Cross-Linked CpG-Oligonucleotides in ß-Glucan Nanogel and Its Antitumor Effect.

Cancer vaccine has the ability to directly eradicate tumor cells by creating and activating cytotoxic T lymphocytes (CTLs). To achieve efficient CTL activity and to induce Th1 responses, it is essential to administer an appropriate adjuvant as well as an antigen. CpG-ODN is known as a ligand of Toll-like receptor 9 (TLR9) and strongly induces Th1 responses. In our previous study, we developed a CpG-ODN delivery system by use of the formation of complexes between ODN and a ß-glucan SPG, denoted as CpG/SPG, and demonstrated that CpG/SPG induces high Th1 responses. In this study, we created a nanogel made from CpG/SPG complexes through DNA-DNA hybridization (cross-linked (CL)-CpG). Immunization with CL-CpG induced much stronger antigen-specific Th1 responses in combination with the antigenic protein ovalbumin (OVA) than that with CpG/SPG. Mice preimmunized with CL-CpG and OVA exhibited a long delay in tumor growth and an improved survival rate after tumor inoculation. These immune inductions can be attributed to the improvement of cellular uptake by the combination of increased size and the cluster effect of the ß-glucan recognition site in the nanogel structure. In other words, the particle nature of CL-CpG, instead of the semiflexible rod conformation of CpG/SPG, enhanced the efficacy of a cancer vaccine. The present results indicate that CL-CpG can be used as a potent vaccine adjuvant for the treatment of cancers and infectious diseases.

Gene silencing using a conjugate comprising Tat peptide and antisense oligonucleotide with phosphorothioate backbones.

Antisense oligonucleotides (ASOs) have a great therapeutic potential for the modulation of gene expression because of the high specificity. The major obstacles for clinical application are enzymatic degradation and low uptake into cells in vivo. In this study, we prepared the conjugate comprising Tat peptide and ASO with phosphorothioate linkages in a simple manner; azide alkyne Huisgen cycloaddition using a copper catalyst. The obtained conjugate showed a high stability in serum, compared with the conjugate with phosphodiester linkages. The conjugates with antisense for c-myb that is transcriptional factor concerning cell growth inhibited the cell proliferation in a dose dependent manner sequence-specifically. These findings suggest Tat-mediated ASOs delivery is useful for the treatment of various diseases.

Characterization of disease-specific alterations in metabolites and effects of mesenchymal stromal cells on dystrophic muscles.

Introduction: Duchenne muscular dystrophy (DMD) is a genetic disorder caused by mutations in the dystrophin-encoding gene that leads to muscle necrosis and degeneration with chronic inflammation during growth, resulting in progressive generalized weakness of the skeletal and cardiac muscles. We previously demonstrated the therapeutic effects of systemic administration of dental pulp mesenchymal stromal cells (DPSCs) in a DMD animal model. We showed preservation of long-term muscle function and slowing of disease progression. However, little is known regarding the effects of cell therapy on the metabolic abnormalities in DMD. Therefore, here, we aimed to investigate the mechanisms underlying the immunosuppressive effects of DPSCs and their influence on DMD metabolism. Methods: A comprehensive metabolomics-based approach was employed, and an ingenuity pathway analysis was performed to identify dystrophy-specific metabolomic impairments in the mdx mice to assess the therapeutic response to our established systemic DPSC-mediated cell therapy approach. Results and Discussion: We identified DMD-specific impairments in metabolites and their responses to systemic DPSC treatment. Our results demonstrate the feasibility of the metabolomics-based approach and provide insights into the therapeutic effects of DPSCs in DMD. Our findings could help to identify molecular marker targets for therapeutic intervention and predict long-term therapeutic efficacy.

Innsbruck-style Retromandibular Anterior Trans-parotid Approach for Condylar Fractures: A Retrospective Review of 39 Fractures.

The retromandibular anterior trans-parotid (RAT) approach and a triangular-positioned double mini-plate osteosynthesis (TDO) technique have been reported from Innsbruck Medical University. This minimally invasive technique involves direct visualization of the condyle and is associated with lower incidence of facial palsy. Methods: A retrospective review was performed on the RAT approach and TDO technique conducted by a surgeon and team at two hospitals in Tokyo during a period of 3 years and 10 months. Results: This technique was performed on 35 patients with 39 condylar fractures. Sixty-nine percent of cases were due to accidental fall, 17% to traffic accidents, and 9% to sports. Furthermore, 92% cases were condylar base fractures. Nighty-seven percent of cases achieved good occlusion. The mean maximum mouth opening was 49 ± 1.3 mm. Postoperatively, facial palsy developed in three patients (7.7%), and two of them developed Frey syndrome at approximately 2.5 years postoperatively (5.1%). All patients completely recovered within 3 months postoperatively. One case each of salivary fistula, visible scar, and condylar resorption was found (2.6%). No case of massive bleeding during surgery, hematoma, or TMJ pain after surgery was found. Conclusion: This technique could achieve good occlusion with low incidence of complications and could contribute to early social reintegration among patients.

[Five-year visual outcomes of typical age-related macular degeneration and/or polypoidal choroidal vasculopathy patients who received photodynamic therapy (PDT) as initial treatment in comparison with patients prior to the PDT era].

PURPOSE: To evaluate five-year visual outcomes of typical age-related macular degeneration (AMD) and polypoidal choroidal vasculopathy (PCV) in patients who received photodynamic therapy (PDT) as initial treatment compared with the outcomes of patients prior to the PDT era. SUBJECTS AND METHODS: Twenty-three eyes observed for 5 years before PDT was available (group A: typical AMD/PCV 16 eyes/7 eyes) and 61 eyes which had been observed for 5 years after PDT with additional treatment as needed (group B: typical AMD/PCV 25 eyes/36 eyes). The visual changes in these groups were retrospectively compared. RESULTS: In group A of typical AMD patients, the mean visual acuity (VA, logMAR) was significantly worse at the 3-year visit and later. In group B of typical AMD patients, the VA was stabilized after 2 years and no significant mean VA deterioration was observed for 5 years. More patients in group B retained a logMAR of less than 1.0 (43% vs. 25%) than in group A. Those patients in group B with PCV, maintained the VA for one year, but it gradually worsened thereafter. CONCLUSION: The PDT shortened the duration of VA deterioration in typical AMD patients from 5 to 2 years with no significant VA decrease for 5 years. The positive effect of PDT on PCV eyes was temporary.

Oligonucleotide delivery to antigen presenting cells by using schizophyllan.

Schizophyllan (SPG), a member of the ß-glucan family, can form novel complexes with homo-polynucleotides such as poly(dA) through hydrogen bonding between two main chain glucoses and the one nucleotide base. Dectin-1, one of the major receptors for ß-glucans, is known to be expressed on antigen presenting cells (APCs) such as macrophages and dendritic cells. This suggests that the above-mentioned complexes could deliver bound functional oligonucleotides (ODNs) including antisense (AS)-ODNs, small interfering RNA, and CpG-ODNs to the APCs. Analysis using a quartz crystal microbalance revealed that a complex consisting of SPG and dA60 with a phosphorothioate backbone was recognized by recombinant Dectin-1 protein. Treatment with this complex containing an AS-ODN for tumor necrosis factor alpha protected mice against lipopolysaccharide-induced hepatitis at a very low AS-ODN dose. Moreover, immunization with CpG-ODN/SPG complex and antigenic proteins induced potent antigen specific immune responses. The present review also represents peptide delivery by conjugation with dA60 and the preparation of a nanogel using DNA-DNA hybridization. These findings indicate that the delivery of a specific ODN using ß-glucans could be used for treating various diseases caused by APCs and for activating antigen specific immune responses.

Intraocular gas dynamics after 20-gauge and 23-gauge vitrectomy with sulfur hexafluoride gas tamponade.

PURPOSE: The purpose of this study was to evaluate the intraocular gas dynamics after 23-gauge transconjunctival sutureless vitrectomy (TSV) as compared with 20-gauge pars plana vitrectomy (PPV). METHODS: A consecutive series of 290 eyes that experienced 20-gauge or 23-gauge vitrectomy with 25% sulfur hexafluoride (SF6) gas tamponade were retrospectively reviewed. Intraocular gas bubble size on postoperative Day 1 and Gas50, the interval to dissipate to a 50% gas fill, were evaluated. RESULTS: The mean intraocular bubble size on postoperative Day 1 was 92.0 ± 8.3% in the 20-gauge PPV cases and 83.8 ± 13.7% in the 23-gauge TSV cases (P < 0.001). The mean Gas50 was 8.6 ± 1.6 days in the 20-gauge PPV cases and 6.6 ± 2.2 days in the 23-gauge TSV cases (P < 0.001). Thorough peripheral vitrectomy and 23-gauge TSV were significantly associated with Gas50 ≤ 4 days (odds ratio, 4.62 and 16.8; P = 0.036 and P = 0.007, respectively). Among thoroughly vitrectomized eyes, 13 eyes treated with 23-gauge PPV with intraoperative suture placement at the sclerotomy sites had gas longevity comparative to those with 20-gauge PPV. CONCLUSION: Eyes treated with 23-gauge TSV tend to have earlier gas disappearance or incomplete gas fill. Intraoperative suture placement would be a solution.

Practical and reliable synthesis of 2',3',5',5″-tetradeuterated uridine derivatives.

Deuterated drugs are valuable in the fields of drug discovery and medicinal chemistry. 2',3',5',5″-tetradeuterated uridine derivatives were synthesised from 2,3,5,5'-selectively tetradeuterated ribose using Sajiki's H-D exchanged Ru/C-H2-D2O-NaOH system and silyl-Hilbert-Johnson methods. The total deuterium content of the tetradeuterated uridines was over 92% using either basic or acidic reaction conditions. These derivatives would be expected as building blocks for the synthesis of deuterium-substituted nucleic acid probes for tracking the pharmacokinetics of nucleic acid drugs.

Polarization-sensitive optical coherence tomography for estimating relative melanin content of autologous induced stem-cell derived retinal pigment epithelium.

Transplantation of autologous human induced pluripotent stem cell-derived retinal pigment epithelial (hiPSC-RPE) sheets is a promising therapy for age-related macular degeneration (AMD). As melanin content is a representative feature of healthy RPE, we used polarization-sensitive optical coherence tomography (PS-OCT) to estimate the relative melanin content of RPE in diseased and non-diseased area, and in human iPSC-RPE sheets in vitro and in vivo by evaluating the randomness of polarization (entropy). Two aged Japanese women, one with neovascular AMD that underwent transplantation of an autologous hiPSC-RPE cell sheet and another with binocular dry AMD, were selected for this study. Entropy value was minimal in cells containing no melanin, whereas that of human RPE and hiPSC-RPE sheets was high. En face entropy of the cultured hiPSC-RPE sheet was compared with its grey-scale photo and its values were found to be inversely correlated with the extent of absence of pigmentation in vitro. En face entropy maps were compared to colour fundus photographs, fundus autofluorescence images, and fluorescein angiography images from patients. Entropy values of intact and defective RPEs and of iPSC-RPE transplant areas were determined in vivo using PS-OCT B-scan images. PS-OCT was found to be applicable in the estimation of relative melanin content of cultured and transplanted RPEs in regenerative medicine.

Melanin concentration and depolarization metrics measurement by polarization-sensitive optical coherence tomography.

Imaging of melanin in the eye is important as the melanin is structurally associated with some ocular diseases, such as age-related macular degeneration. Although optical coherence tomography (OCT) cannot distinguish tissues containing the melanin from other tissues intrinsically, polarization-sensitive OCT (PS-OCT) can detect the melanin through spatial depolarization of the backscattered light from the melanin granules. Entropy is one of the depolarization metrics that can be used to detect malanin granules in PS-OCT and valuable quantitative information on ocular tissue abnormalities can be retrived by correlating entropy with the melanin concentration. In this study, we investigate a relationship between the melanin concentration and some depolarization metrics including the entropy, and show that the entropy is linearly proportional to the melanin concentration in double logarithmic scale when noise bias is corrected for the entropy. In addition, we also confirm that the entropy does not depend on the incident state of polarization using the experimental data, which is one of important attributes that depolarization metrics should have. The dependence on the incident state of polarization is also analyzed for other depolarization metrics.

Long-term results of photodynamic therapy or ranibizumab for polypoidal choroidal vasculopathy in LAPTOP study.

BACKGROUND/AIM: We previously reported that ranibizumab performed better on visual prognosis than photodynamic therapy (PDT) in a Ranibizumab (Lucentis) And Photodynamic Therapy On Polypoidal choroidal vasculopathy (LAPTOP) study. To determine if the first-choice treatment, either PDT or intravitreal ranibizumab, has a long-term effect in patients with polypoidal choroidal vasculopathy (PCV). METHODS: We reviewed medical records of patientsrandomised to either PDT (29 eyes) or ranibizumab (27 eyes) from July 2009 to June 2011 in LAPTOP study. Retreatment or switching to other treatments were at the investigator's discretion after release from the 2-year LAPTOP study up to 5years. We evaluated visual acuity (VA), continuity of initial treatment, percentage of dry macula achievement and macular atrophy at 5 years. RESULTS: The logarithm of minimal angle of resolution VA was 0.56 in the PDT and 0.44 in the ranibizumab groups at baseline (p=0.101) and was 0.55 and 0.28 at 5years, respectively (p<0.05). More than 70% of the patients converted to aflibercept in following years. Achievement percentages of dry macula were 74% (PDT) and 63% (ranibizumab) at 5years, and macular atrophy was detected in 78% (PDT) and 60% (ranibizumab) with a mean area of 7.7 and 3.5 mm2, respectively (p=0.155). CONCLUSIONS: The better VA in the initial ranibizumab group compared with the PDT group at 2 years was retained at the 5-year follow-up.

Physiological responses of simulated karate sparring matches in young men and boys.

The purpose of this study was to investigate the duration of each series of offensive and defensive techniques and the cardiovascular, metabolic, and perceptual responses during 2- and 3-minute bouts of simulated karate sparring. Six young men (age, 18-20 years) and 6 boys (age, 16-17 years) participated in this study. We formed 3 pairs of men and 3 pairs of boys to create a demanding competitive environment. After a rest period, each pair performed a 2-minute bout of sparring, sat quietly for 60 minutes, and then performed 3-minute bout of sparring. We measured oxygen uptake (Vo2), heart rate (HR), and blood lactate responses and ascertained the rate of perceived exertion (RPE) and energy expenditure (EE) during these sparring bouts. The ventilatory threshold was estimated from ventilatory equivalent and Vo2 obtained during the treadmill test. The duration of each series of offensive and defensive techniques was videotaped. During the 2- and 3-minute bouts of sparring, the duration of longest series of offensive and/or defensive combination techniques performed were 2.1 +/- 1.0 and 1.8 +/- 0.4 seconds, respectively; the mean total times of performing offensive and defensive techniques were 13.3 +/- 3.3 and 19.4 +/- 5.5 seconds, respectively. The mean oxygen uptake (Vo2), the percentage of maximum oxygen uptake (%Vo2max), HR, percentage of maximum HR, RPE, and EE for a 3-minute bout of sparring were significantly higher than for a 2-minute bout of sparring. The mean %Vo2max values for these bouts of sparring were below the ventilatory threshold. It is recommended that karate practitioners perform more specific weight training, plyometric exercises, and interval training to increase the ability to buffer acid muscle and blood concentrations and to build lean body mass, strength, and power to develop the specific motor skills required in sparring.

Designing an immunocyte-targeting delivery system by use of beta-glucan.

A ß-1,3-d-glucan called Schizophyllan (SPG) can form a novel complex with homo oligodeoxynucleotides (ODNs) via the combination of hydrogen bonding and hydrophobic interactions. Dectin-1 is a major receptor involved in the recognition of ß-1,3-d-glucans and expressed on antigen presenting cells (APCs) including macrophages, dendritic cells, monocytes, neutrophils, and a subset of T cells. Therefore, the SPG/ODN complex can be used as APCs cell-specific delivery of functional ODNs including unmethylated CpG sequences (CpG-ODNs). In fact, CpG-ODN/SPG complex induced high antibody titers when it was administered to cynomolgus monkeys as adjutant of influenza vaccine. These results indicate that SPG can be an excellent immunocyte-targeting drug delivery system.

Restoration of Photoreceptor Outer Segments up to 24 Months after Pars Plana Vitrectomy in Patients with Diabetic Macular Edema.

PURPOSE: To evaluate the ellipsoid zone (EZ) and external limiting membrane (ELM) status on optical coherence tomography images in patients with diabetic macular edema (DME) before and after pars plana vitrectomy (PPV). DESIGN: Retrospective observational analysis. PARTICIPANTS: Sixty-one eyes of 49 patients with DME. METHODS: We retrospectively reviewed the spectral-domain optical coherence tomography images of 61 eyes of 49 patients with DME who were treated with PPV and evaluated the percentage disruption of the EZ and ELM before and up to 24 months after PPV. MAIN OUTCOME MEASURES: Longitudinal changes in visual acuity (VA), central retinal thickness (CRT), and EZ and ELM spectral-domain optical coherence tomography findings in patients with DME. RESULTS: The logarithm of the minimum angle of resolution VA was 0.477 before PPV and then improved to 0.372 and 0.344 at 12 and 24 months after PPV, respectively (P < 0.005 and P < 0.001, respectively). The CRT was 505.5 µm before PPV and then decreased gradually to 345.0 and 301.4 µm at 12 and 24 months, respectively (P < 0.001 for both). The percentage disruption of the EZ was 29.3% and ELM was 9.6% before PPV and then deteriorated to 41.6% (P < 0.001) and 15.1% (P < 0.05), respectively, 1 month after PPV. The disruption of the EZ and ELM decreased gradually. The percentage disruption of the ELM recovered but did not improve by 24 months, compared with baseline (11.6%; P = 0.875), whereas the percentage disruption of the EZ recovered to baseline by 6 months (29.4%; P = 1.000) and then decreased significantly at 24 months (21.3%; P < 0.05). CONCLUSIONS: Both the EZ and ELM deteriorated after PPV. Whereas the ELM recovered but did not improve by 24 months when compared with baseline, the EZ and VA improved up to 24 months.

A two-component micelle with emergent pH responsiveness by mixing dilauroyl phosphocholine and deoxycholic acid and its delivery of proteins into the cytosol.

Providing appropriate pH responsiveness for drug delivery nanoparticles is one of the major issues in developing a new generation of delivery systems. This paper reports that, when phosphocholine and a bile acid were mixed, the resultant two-component micelle gained pH responsiveness, while the individual components did not show any such responsiveness. The pH responsiveness was shown to be determined by the chemical structure, especially the positions and chirality of the OH groups, of the bile acid, and the sensitivity was determined by the alkyl chain length of the phosphocholine. The best combination for evading endocytosis was dilauroyl phosphocholine (DLPC) and deoxycholic acid (DA). Small-angle X-ray scattering revealed that the pH responsiveness was related to the change of surface hydrophobicity, namely, decreasing pH led to protonation of the carboxylic acid, resulting in aggregation of the preceding micelles. We assume that particles that become hydrophobic in this way can start interacting with the endocytotic bilayer, which eventually leads to rupture of the endocytotic vesicle. This mechanism is well supported by the finding that fluorescein-conjugated ovalbumin proteins were transported into the cytosol when they were co-administered with DLPC/DA.