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Carbon dioxide-dependent Proteus mirabilis has been isolated from clinical specimens. It is not clear whether mutations in carbonic anhydrase are responsible for the carbon dioxide dependence of P. mirabilis. The pathogenicity of carbon dioxide-dependent P. mirabilis also remains unclear. The purpose of this study was to determine the cause carbon dioxide dependence of P. mirabilis and its pathogenicity. The DNA sequence of can encoding carbonic anhydrase of a carbon dioxide-dependent P. mirabilis small colony variant (SCV) isolate was analyzed. To confirm that impaired carbonic anhydrase activity is responsible for the formation of the carbon dioxide-dependent SCV phenotype of P. mirabilis , we performed complementation experiments using plasmids with intact can. Additionally, mouse infection experiments were performed to confirm the change in virulence due to the mutation of carbonic anhydrase. We found that the can gene of the carbon dioxide-dependent P. mirabilis SCV isolate showed had a frameshift mutation with a deletion of 1 bp (c. 173delC). The can of P. mirabilis encodes carbonic anhydrase was also found to function in Escherichia coli. The cause of the carbon dioxide-dependent SCV phenotype of P. mirabilis was an abnormality in carbonic anhydrase. Nevertheless, no changes were observed in virulence due to the mutation of carbonic anhydrase in mouse infection experiments. The can gene is essential for the growth of P. mirabilis in ambient air. The mechanisms underlying this fitness advantage in terms of infection warrant further investigation. [ABSTRACT FROM AUTHOR]

• Boosting with S-268019-b enhanced immunity against SARS-CoV-2 in macaques. • Squalene-based adjuvant was essential for the booster effect of S-268019-b. • Boosting with S-268019-b enhanced neutralization breadth against Omicron subvariants. • The antigen specific memory B cells expanded after the booster dose of S-268019-b. SARS-CoV-2 Omicron subvariants such as BA.2.12.1, BA.4 and BA.5 have been spreading rapidly and become dominant worldwide. Here we report the homologous or heterologous booster effects of S-268019-b, a recombinant spike protein vaccine with the squalene-based adjuvant A-910823 in cynomolgus macaques. In macaques which had been primed with S-268019-b or mRNA vaccines, boosting with S-268019-b enhanced neutralizing antibodies (NAb) against ancestral SARS-CoV-2. Since boosting with the antigen without adjuvant did not efficiently restore NAb titers, adjuvant A-910823 was essential for the booster effect. Importantly, boosting with S-268019-b enhanced NAb against all of the Omicron subvariants we tested, including BA.2.12.1, BA.4 and BA.5, in comparison to two vaccine doses. Additionally, expansion of Omicron-specific B cells was confirmed after boosting with S-268019-b. These results indicate that a booster dose of S-268019-b with the adjuvant enhances the neutralization breadth. [ABSTRACT FROM AUTHOR]

• A novel SARS-CoV-2 recombinant spike (S)-protein vaccine S-268019-b was developed. • S-268019-b induced antibodies against S-protein and its receptor binding domain. • Neutralizing antibody was induced against SARS-CoV-2 and pseudovirus variants. • S-268019-b demonstrated protective efficacy against SARS-CoV-2 challenge. • S-268019-b was safe for use in cynomolgus macaques. The vaccine S-268019-b is a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S)-protein vaccine consisting of full-length recombinant SARS-CoV-2 S-protein (S-910823) as antigen, mixed with the squalene-based adjuvant A-910823. The current study evaluated the immunogenicity of S-268019-b using various doses of S-910823 and its vaccine efficacy against SARS-CoV-2 challenge in cynomolgus monkeys. The different doses of S-910823 combined with A-910823 were intramuscularly administered twice at a 3-week interval. Two weeks after the second dosing, dose-dependent humoral immune responses were observed with neutralizing antibody titers being comparable to that of human convalescent plasma. Pseudoviruses harboring S proteins from Beta and Gamma SARS-CoV-2 variants displayed approximately 3- to 4-fold reduced sensitivity to neutralizing antibodies induced after two vaccine doses compared with that against ancestral viruses, whereas neutralizing antibody titers were reduced >14-fold against the Omicron variant. Cellular immunity was also induced with a relative Th1 polarized response. No adverse clinical signs or weight loss associated with the vaccine were observed, suggesting safety of the vaccine in cynomolgus monkeys. Immunization with 10 µg of S-910823 with A-910823 demonstrated protective efficacy against SARS-CoV-2 challenge according to genomic and subgenomic viral RNA transcript levels in nasopharyngeal, throat, and rectal swab specimens. Pathological analysis revealed no detectable vaccine-dependent enhancement of disease in the lungs of challenged vaccinated monkeys. The current findings provide fundamental information regarding vaccine doses for human trials and support the development of S-268019-b as a safe and effective vaccine for controlling the current pandemic, as well as general protection against SARS-CoV-2 moving forward. [ABSTRACT FROM AUTHOR]

The usefulness of depression scales for patients with cancer based on item response theory (IRT) and computer adaptive testing (CAT) has not yet been fully explored. This study thus aimed to develop an IRT-based tool for measuring depression in patients with cancer. We analyzed data from 393 patients with cancer from four tertiary centers in Japan who had not received psychiatric treatment. They answered 62 questions across five categories regarding their psychiatric status over the previous week. We selected 28 items that satisfied the assumptions of IRT, fitted a graded response model to these items, and performed CAT simulations. The CAT simulation used an average of 6.96 items and showed a Pearson's correlation coefficient of 0.916 (95% confidence interval, 0.899–0.931) between the degree of depression estimated by simulation and that estimated using all 28 items. The measurement precision of CAT with only four items was superior to that of the estimation using the calibrated Patient Health Questionnaire-9. These results imply that this scale is useful and accurate for measuring depression in patients with cancer. [ABSTRACT FROM AUTHOR]

Background Transforaminal endoscopic lumbar diskectomy (TELD) is a widely used minimally invasive surgical procedure to treat herniated disks. Postoperative dysesthesia (POD) is a significant complication that affects patient satisfaction. Several anatomical and magnetic resonance (MR) imaging studies of the intervertebral foramina showed that TELD should be avoided in patients with small Kambin's triangular working zones. Recently, some reports indicated the usefulness of three-dimensional (3D) computed tomography/magnetic resonance (CT/MR) fusion images. To date, no articles have been published in the English literature regarding the use of 3D CT/MR fusion images before TELD to evaluate Kambin's triangular working zone. Our objective was to examine clinical outcomes when preoperative Kambin's triangular working zone measurements from 3D CT/MR fusion images were used to determine the approach to TELD. Patients and Methods We included 31 patients who underwent TELD. We rotated the 3D CT/MR fusion images from the posteroanterior view on the approach side to the angle at which Kambin's triangular working zone appeared the largest. This was used to determine the intraoperative insertion angle for the working cannula. When the perpendicular line extending from the exiting nerve at the intervertebral disk level to the upper margin of the superior articular process (exiting nerve-superior articular process distance [ESD]) was less than or equal to 7 mm, an approach that combined foraminoplasty with an outside-in technique (F + outside-in) was used. We compared ESD and clinical outcomes, such as POD, between the approaches. Results Surgical plans were based on ESD values from 22 patients. ESD was 7 mm in 21 patients, all of whom underwent F + outside-in. The inside-out approach was used in eight of nine patients who did not have ESD measurements. Of these, five patients had retrospective ESD measurements of 7 mm. The mean ESD was 6.3 ± 1.0 mm when inside-out was used and 4.4 ± 1.6 mm when F + outside-in was used, a significant difference. Significant improvements were observed in the visual analog scale scores for low back pain, lower limb pain, and lower limb paresthesia. There were no incidences of POD or intraoperative complications. Conclusion Based on preoperative Kambin's triangular working zone measurements from 3D CT/MR fusion images, F + outside-in should be selected when the working zone is smaller than the cannula diameter. This method successfully avoided POD in our study. Preoperative Kambin's triangular working zone measurements from 3D CT/MR fusion imaging can enhance patient safety during TELD. [ABSTRACT FROM AUTHOR]

Background: Urinary tract infections (UTIs) are common bacterial infections, primarily caused by uropathogenic Escherichia coli (UPEC), leading to significant health issues and economic burden. Although antibiotics have been effective in treating UPEC infections, the rise of antibiotic-resistant strains hinders their efficacy. Hence, identifying novel bacterial targets for new antimicrobial approaches is crucial. Bacterial factors required for maintaining the full virulence of UPEC are the potential target. MepM, an endopeptidase in E. coli, is involved in the biogenesis of peptidoglycan, a major structure of bacterial envelope. Given that the bacterial envelope confronts the hostile host environment during infections, MepM's function could be crucial for UPEC's virulence. This study aims to explore the role of MepM in UPEC pathogenesis. Results: MepM deficiency significantly impacted UPEC's survival in urine and within macrophages. Moreover, the deficiency hindered the bacillary-to-filamentous shape switch which is known for aiding UPEC in evading phagocytosis during infections. Additionally, UPEC motility was downregulated due to MepM deficiency. As a result, the mepM mutant displayed notably reduced fitness in causing UTIs in the mouse model compared to wild-type UPEC. Conclusions: This study provides the first evidence of the vital role of peptidoglycan endopeptidase MepM in UPEC's full virulence for causing UTIs. MepM's contribution to UPEC pathogenesis may stem from its critical role in maintaining the ability to resist urine- and immune cell-mediated killing, facilitating the morphological switch, and sustaining motility. Thus, MepM is a promising candidate target for novel antimicrobial strategies. [ABSTRACT FROM AUTHOR]

Background With effective preoperative planning, percutaneous endoscopic diskectomy (PED) via an interlaminar approach (PED-IL) can reduce the need for partial laminectomy. Our aim was to assess the clinical outcomes of PED-IL, planned using three-dimensional fusion of computed tomography and magnetic resonance (3D CT/MR) images. Material and Methods Our retrospective analysis was based on data from 102 patients (66 were men) treated by PED-IL. Preoperative planning was based on the positional relationship between the nerve root and the lumbar disk herniation, visualized on 3D CT/MR fusion images through a simulated IL window. Two approaches were planned: type I, via a partial window opening of the ligamentum flavum, and type II, via a partial IL window opening and partial laminectomy. Decisions were then made to approach the herniation from the shoulder (subtype s) or axillary (subtype a) portion of the nerve, or we used a combination of both approaches (subtype a + s). Operative time and the pre- to postsurgical change in pain and pain-related disability scores (Japanese Orthopaedic Association Back Pain Evaluation Questionnaire scores) were evaluated. The number of cases in which the preoperative planned surgery was modified intraoperatively and the incidence of complications were also evaluated. Results A type I approach was planned in 30 cases (29.4%) and type II in 72 cases (70.6%). Intraoperative change from type I to type II was required in two cases. The mean operative time was longer for type II than type I procedures. Pain and pain-related disability scores improved in all patients, and only one case of dural sleeve damage was identified. Conclusions Preoperative planning using a simulated IL window to visualize the 3D regional anatomy is effective in predicting the feasibility of a PED-IL. [ABSTRACT FROM AUTHOR]

Pseudoaneurysm is one of the most serious complications of femoral trochanteric fracture surgery. Since the rupture of pseudoaneurysm may lead to death, early detection is important. We report the case of an 80-year-old male who developed pain in the proximal thigh and severe swelling after internal fixation of a femoral trochanteric fracture with a hip nail. Angiography revealed a pseudoaneurysm of a branch of the deep femoral artery near the interlocking screw. The vascular lesion was immediately treated by transcatheter embolization, and the vascular leakage was completely resolved with catheter embolization. After embolization, the patient’s clinical state improved rapidly, and the laboratory values improved to normal after several weeks. The current case study reminds us that pseudoaneurysm can occur after intramedullary nail placement to treat a femoral trochanteric fracture. [ABSTRACT FROM AUTHOR]

Purpose: To assess the feasibility of Four-Dimensional Ultra-Low-Dose Computed Tomography (4D-ULDCT) for distinguishing pleural aspects with localized pleural adhesion (LPA) from those without.Methods: Twenty-seven patients underwent 4D-ULDCT during a single respiration with a 16cm-coverage of the body axis. The presence and severity of LPA was confirmed by their intraoperative thoracoscopic findings. A point on the pleura and a corresponding point on the outer edge of the costal bone were placed in identical axial planes at end-inspiration. The distance of the two points (PCD), traced by automatic tracking functions respectively, was calculated at each respiratory phase. The maximal and average change amounts in PCD (PCDMCA and PCDACA) were compared among 110 measurement points (MPs) without LPA, 16MPs with mild LPA and 10MPs with severe LPA in upper lung field cranial to the bronchial bifurcation (ULF), and 150MPs without LPA, 17MPs with mild LPA and 9MPs with severe LPA in lower lung field caudal to the bronchial bifurcation (LLF) using the Mann-Whitney U test.Results: In the LLF, PCDACA as well as PCDMCA demonstrated a significant difference among non-LPA, mild LPA and severe LPA (18.1±9.2, 12.3±6.2 and 5.0±3.3mm) (p<0.05). Also in the ULF, PCDACA showed a significant difference among three conditions (9.2±5.5, 5.7±2.8 and 2.2±0.4mm, respectively) (p<0.05), whereas PCDMCA for mild LPA was similar to that for non-LPA (12.3±5.9 and 17.5±11.0mm).Conclusions: Four D-ULDCT could be a useful non-invasive preoperative assessment modality for the detection of the presence or severity of LPA. [ABSTRACT FROM AUTHOR]

The Da Vinci Surgical System is an ergonomically devised and excellent surgical support device. However, surgeon skill is of paramount importance since human error cannot be completely eliminated. We report a case of bleeding from the pulmonary artery due to a footswitch misstep. A 72-year-old male with suspected right upper lobe lung cancer underwent robot-assisted thoracoscopic surgery (RATS). While avoiding the pulmonary artery with the right arm spatula and trying to cauterize V2t with the left arm bipolar-forceps, the footswitch was accidently activated and the spatula was energized, resulting in pulmonary artery trauma and blood loss. After this case, we changed the surgical procedure from a monopolar–bipolar combination use to a bipolar-only use and noted no significant difference in the console duration, and less intraoperative blood loss. Human errors can occur anytime. Especially for surgeons new to RATS, simplified foot management should be considered until RATS mastery is achieved. [ABSTRACT FROM AUTHOR]

This document is a correction notice for an article titled "Immune response and protective efficacy of the SARS-CoV-2 recombinant spike protein vaccine S-268019-b in mice" published in Scientific Reports. The correction addresses an error in Figure 6 of the original article, where panel (e) was omitted. The corrected figure and accompanying legend are provided in the notice. The original article has been updated to reflect this correction. [Extracted from the article]

The Ygj D protein is essential for the synthesis of the universal t RNA modification, N6-threonylcarbamoyladenosine (t6 A), which is necessary for the decoding of ANN codons. We isolated a suppressor ( ygj Dsup) of the ygj Dts mutant by its permissive growth at high temperature in Escherichia coli. Resequencing of the ygj Dsup mutant genome showed the presence of a complicated chromosome rearrangement, an inverse insertion of a large duplicated region ( c. 450 kb) into a small deleted region. The temperature-resistant growth associated with ygj Dsup was due to the presence of multicopy suppressor genes, yjeE and groL, of the ygj Dts mutation in the duplicated region. This DNA rearrangement was not simply mediated by IS 1 transposition, but the duplicated region was flanked by IS 1. We showed that the frequency of IS 1 transposition was increased in ygj Dts mutants. The transposase of IS 1 is coded for by the insB gene, and its translation occurs through a frameshift of a ribosome translating upstream of the insA gene. We showed that this frameshifting frequency was increased by the ygj Dts mutation. These results indicated that the mutation of the gene for t RNA modification, t6 A, affected IS 1 transposition. [ABSTRACT FROM AUTHOR]

The riboflavin analog roseoflavin is an antibiotic produced by Streptomyces davawensis. Riboflavin transporters are responsible for roseoflavin uptake by target cells. Roseoflavin is converted to the flavin mononucleotide (FMN) analog roseoflavin mononucleotide (RoFMN) by flavokinase and to the flavin adenine dinucleotide (FAD) analog roseoflavin adenine dinucleotide (RoFAD by FAD synthetase. In order to study the effect of RoFMN and RoFAD in the cytoplasm of target cells, Escherichia coli was used as a model. E. coli is predicted to contain 38 different FMN- or FAD-dependent proteins (flavoproteins). These proteins were overproduced in recombinant E. coli strains grown in the presence of sublethal amounts of roseoflavin. The flavoproteins were purified and analyzed with regard to their cofactor contents. It was found that 37 out of 38 flavoproteins contained either RoFMN or RoFAD. These cofactors have different physicochemical properties than FMN and FAD and were reported to reduce or completely abolish flavoprotein function. [ABSTRACT FROM AUTHOR]

Bacterial peptidoglycan acts as an exoskeleton to protect the bacterial cell. Although peptidoglycan biosynthesis by penicillin- binding proteins is well studied, few studies have described peptidoglycan disassembly, which is necessary for a dynamic structure that allows cell growth. In Bacillus subtilis, more than 35 genes encoding cell wall lytic enzymes have been identified; however, only two D,L-endopeptidases (lyre and cwlO) are involved in cell proliferation. In this study, we demonstrated that the D,L-endopeptidase activity at the lateral cell wall is essential for cell proliferation. Inactivation of LytE and CwlO by point mutation of the catalytic residues caused cell growth defects. However, the forced expression of LytF or CwlS, which are paralogs of LytE, did not suppress lyre cwlO synthetic lethality. Subcellular localization studies of these D,L-endopeptidases showed LytF and CwlS at the septa and poles, CwlO at the cylindrical part of the cell, and LytE at the septa and poles as well as the cylindrical part. Furthermore, construction of N-terminal and C-terminal domain-swapped enzymes of LytE, LytF, CwlS, and CwlO revealed that localization was dependent on the N-terminal domains. Only the chimeric proteins that were enzymatically active and localized to the sidewall were able to suppress the synthetic lethality, suggesting that the lack of D,L-endopeptidase activity at the cylindrical part of the cell leads to a growth defect. The functions of LytE and CwlO in cell morphogenesis were discussed. [ABSTRACT FROM AUTHOR]

Suppressor of cytokine signaling-1 (SOCS1) has been shown to be an essential negative regulator of cytokine responses, including those of IFNγ, IL-2, IL-4 and IL-7. SOCS1 deficiency resulted in hyperactivation not only of T cells in general but also of NKT cells specifically. Consistent with previous reports, T- and NKT-cell-specific deletion of Socs1 in mice resulted in enhanced sensitivity to ConA-induced hepatitis. Compared with wild-type (WT) NKT cells, SOCS1-deficient NKT cells produced larger quantities of IFNγ in response to ConA and proliferated faster in response to IL-2 and IL-15. To our surprise, however, SOCS1-deficient NKT cells did not respond to the synthetic glycolipid ligand alpha-galactosylceramide (α-GalCer), though they did respond to sulfatide. α-GalCer-CD1d-tetramer-positive type I NKT [invariant NKT (iNKT)] cells were marginally detected in the periphery of SOCS1-conditional knockout (cKO) mice, suggesting that most of the SOCS1-deficient NKT cells at the periphery were type II NKT cells. Consistently, invariant Vα14 expression was much lower in SOCS1-deficient NKT cells than in WT NKT cells, indicating that iNKT cell homeostasis was abnormal in SOCS1-cKO mice. This reduction in iNKT cells was not observed in mice of an IFNγ-deficient background. These results suggest that SOCS1 is an important regulator of the balance between type I and type II NKT cells at the periphery. [ABSTRACT FROM AUTHOR]

Abstract: In light of reports that the administration of fasudil, a Rho-kinase inhibitor, improved rats locomotor abilities following spinal cord injury, we hypothesized that combining fasudil with another type of therapy, such as stem cell transplantation, might further improve the level of locomotor recovery. Bone marrow stromal cells (BMSCs) are readily available for stem cell therapy. In the present study, we examined whether fasudil combined with BMSC transplantation would produce synergistic effects on recovery. Adult female Sprague–Dawley rats were subjected to spinal cord contusion injury at the T10 vertebral level using an IH impactor (200 Kdyn). Immediately after contusion, they were administrated fasudil intrathecally for 4 weeks. GFP rat-derived BMSCs (2.5×106) were injected into the lesion site 14 days after contusion. Locomotor recovery was assessed for 9 weeks with BBB scoring. Sensory tests were conducted at 8 weeks. Biotinylated dextran amine (BDA) was injected into the sensory-motor cortex at 9 weeks. In addition to an untreated control group, the study also included a fasudil-only group and a BMSC-only group in order to compare the effects of combined therapy vs. single-agent therapy. Animals were perfused transcardially 11 weeks after contusion, and histological examinations were performed. The combined therapy group showed statistically better locomotor recovery than the untreated control group at 8 and 9 weeks after contusion. Neither of the two single-agent treatments improved open field locomotor function. Sensory tests showed no statistically significant difference by treatment. Histological and immunohistochemical studies provided some supporting evidence for better locomotor recovery following combined therapy. The average area of the cystic cavity was significantly smaller in the fasudil+BMSC group than in the control group. The number of 5-HT nerve fibers was significantly higher in the fasudil+BMSC group than in the control group on the rostral side of the lesion site. BDA-labeled fibers on the caudal side of the lesion epicenter were observed only in the fasudil+BMSC group. On the other hand, only small numbers of GFP-labeled grafted cells remained 9 weeks after transplantation, and these were mainly localized at the site of injection. Double immunofluorescence studies showed no evidence of differentiation of grafted BMSCs into glial cells or neurons. The Rho-kinase inhibitor fasudil combined with BMSC transplantation resulted in better locomotor recovery than occurred in the untreated control group. However, the data failed to demonstrate significant synergism from combined therapy compared with the levels of recovery following single-agent treatment. [Copyright &y& Elsevier]

Th cells have long been divided into two subsets, Th1 and Th2; however, recently, Th17 and inducible regulatory T (iTreg) cells were identified as new Th cell subsets. Although Th1- and Th2-polarizing cytokines have been shown to suppress Th17 and iTreg development, transcriptional regulation of Th17 and iTreg differentiation by cytokines remains to be clarified. In this study, we found that expression of the growth factor independent 1 (Gfi1) gene, which has been implicated in Th2 development, was repressed in Th17 and iTreg cells compared with Th1 and Th2 lineages. Gfi1 expression was enhanced by the IFN-γ/STAT1 and IL-4/STAT6 pathways, whereas it was repressed by the transforming growth factor-β1 stimulation at the promoter level. Over-expression of Gfi1 strongly reduced IL-17A transcription in the EL4 T cell line, as well as in primary T cells. This was due to the blockade of recruitment of retinoid-related orphan receptor γt to the IL-17A promoter. In contrast, IL-17A expression was significantly enhanced in Gfi1-deficient T cells under Th17-promoting differentiation conditions as compared with wild-type T cells. In contrast, the impacts of Gfi1 in iTregs were not as strong as in Th17 cells. Taken together, these data strongly suggest that Gfi1 is a negative regulator of Th17 differentiation, which represents a novel mechanism for the regulation of Th17 development by cytokines. [ABSTRACT FROM PUBLISHER]

Background: Placement of aortic stent-valves in the proper position is difficult due to the high velocity and oscillation of blood flow and anatomical characteristics. To address this problem, a re-positionable stent-valve was designed and examined in a preliminary animal experiment. Methods: Protocols for this animal study were approved by the institutional Animal Care and Use Committee. An umbrella-shaped polyurethane valve was attached at the tip of newly devised re-positionable metallic stent. The prosthesis (re-positionable stent-valve) was implanted in 5 female pigs via the right carotid artery using a 10-F introducer (Group A). As a control, the stent-without-valve was implanted in 4 female pigs (Group B). Efficacy of the valve was assessed by aortography and aortic pressure measurement. Animals were sacrificed 1 hour after stent-valve implantation for macroscopic examination. Results: The stent-valve was successfully implanted in all pigs. In Group A, aortography showed good function of the polyurethane valve. No significant change in diastolic pressure was noted during valve testing. In Group B, aortography and aortic pressure measurement showed massive aortic regurgitation following dysfunction of the native aortic leaflets. Post-mortem examination showed small amounts of thrombus inside the polyurethane umbrella in Group A. Conclusion: Placement of the re-positionable stent-valve seems feasible and effective. However, the problem of thrombus formation inside the umbrella valve should be solved in future studies. [ABSTRACT FROM AUTHOR]

Threed,l-endopeptidases, LytE, LytF and CwlS, are involved in the vegetative cell separation in Bacillus subtilis. A novel cell surface protein, IseA, inhibits the cell wall lytic activities of thesed,l-endopeptidases in vitro, and IseA negatively regulates the cell separation enzymes at the post-translational level. Immunofluorescence microscopy indicated that the IseA−3xFLAG fusion protein was specifically localized at cell separation sites and poles on the vegetative cell surface in a similar manner of thed,l-endopeptidases. Furthermore, pull-down assay showed that IseA binds to the catalytic domain of LytF, indicating that IseA is localized on the cell surface through the catalytic domain of LytF. Overexpression of IseA caused a long-chained cell morphology in the exponential growth phase, indicating that IseA inhibits the cell separationd,l-endopeptidases in vivo. Besides, overexpression of IseA in a cwlO disruptant affected cell growth, implying that IseA is also involved in the cell elongation event. However, although IseA inhibits the activities of LytE, LytF, CwlS and CwlO in vitro, it is unlikely to inhibit CwlS and CwlO in vivo. This is the first demonstration that the cell separation event is post-translationally controlled through a direct interaction between cell separation enzymes and a specific novel inhibitor in bacteria. [ABSTRACT FROM AUTHOR]

Osteopontin (OPN) is expressed in many tissues during inflammatory responses. After spinal cord injury, microglia expresses OPN at the site of injury during the early to subacute stages. However, the function of OPN in spinal cord injury is not well understood. This study examines the responses of OPN knock-out (KO) and wild-type (WT) mice to spinal cord contusion injury. KO and WT mice were injured with a modified New York University impactor. Weights of 10 or 5.6 g were dropped 6.25 mm onto the T13 spinal cord under isoflurane anesthesia. At 24 h, homogenized spinal cords were analyzed for total potassium concentration to estimate lesion volumes. Expression of apoptotic genes, proinflammatory cytokines, and nerve growth factors was measured by reverse transcription (RT)- PCR and Western blot. In a series of animals, locomotor recovery was assessed with the Basso mouse scale (BMS) for 6 weeks, and histological analyses was performed to determine tissue preservation. Lesion volume showed no significant differences between KO and WT mice at 24 h. RT-PCR indicated that KO mice had significantly less Bcl- 2, tumor necrosis factor-α, interleukin-1β, and interleukin-6 mRNA compared with WT controls. Western blot also showed that KO had significantly less Bcl-27 d after spinal cord injury. KO mice had significantly worse BMS locomotor scores than WT at 6 weeks. KO mice also had a significantly reduced area of spared white matter and fewer neuronal-specific nuclear protein-positive neurons in the spinal cord surrounding the impact site. This result supports a potential neuroprotective role for OPN in the inflammatory response to spinal cord injury. [ABSTRACT FROM AUTHOR]

A carbaryl hydrolase gene (cahA) encoded on the plasmid pRC1 in Arthrobacter sp. RC100 was cloned and sequenced. The entire region of the deduced amino acid sequence was found to be homologous to that of an amidase family. Parts of the consensus sequences of the amidase gene have been identified in CahA from strain RC100. CahA was overexpressed in Escherichia coli JM109, and the enzyme was purified to homogeneity by protamine sulfate treatment, ammonium sulfate precipitation, and hydrophobic and anion-exchange chromatographies. The purified enzyme showed hydrolase activity toward 1-naphthylacetamide and isobutyramide but showed no activity toward 1-naphthylacetate. This is the first report of an amidase that is able to hydrolyze N-methylcarbamate pesticides. [Copyright &y& Elsevier]

The Escherichia coli hda gene codes for a DnaA-related protein that is essential for the regulatory inactivation of DnaA (RIDA), a system that controls the initiation of chromosomal replication. We have identified the ygfZ gene, which encodes a folate-binding protein, as a suppressor of hda mutations. The ygfZ null mutation suppresses an hda null mutation. The over-initiation and abortive elongation phenotypes conferred by the hda mutations are partially suppressed in an hda ygfZ background. The accumulation of the active form of DnaA, ATP-DnaA, in the hda mutant is suppressed by the disruption of the ygfZ gene, indicating that YgfZ is involved in regulating the level of ATP-DnaA. Although ygfZ is not an essential gene, the ygfZ disruptant grows slowly, especially at low temperature, demonstrating that this gene is important for cellular proliferation. We have identified mnmE ( trmE) as a suppressor of the ygfZ disruption. This gene encodes a GTPase involved in tRNA modification. Examination of RNA modification in the ygfZ mutant reveals reduced levels of 2-methylthio-6-iodeadenosine, indicating that YgfZ participates in the methylthio-group formation of this modified nucleoside in some tRNAs. These results suggest that YgfZ is a key factor in regulatory networks that act via tRNA modification. [ABSTRACT FROM AUTHOR]

The purpose of this study was to use a cDNA microarray to identify new genes involved in healing of spinal cord injury. C57BL/6 mice (7-8 weeks, male) were subjected to spinal cord compression injury (SCI) at the T7/8 level (20 g, 5 min; SCI group). For the control group, mice underwent only laminectomy. Mice were killed at 1, 3 and 7 days. cDNA transcribed from mRNA was hybridized to NIA mice 15K microarrays at each time point. We found 84 genes showing significant expressional changes, including higher and lower expression levels in the SCI groups than in the control [more than 1.0 or less than -1.0 using log ratio (base 2)]. Five genes were selected for further quantitative gene expression analysis by real-time reverse transcription (RT)-PCR. For histological examination, we applied in situ hybridization and fluorescence immunohistochemistry. Cathepsin D, metallothionein-1 (MT-1), metallothionein-2 (MT-2), osteopontin (OPN), and tenascin-C were selected for quantitative and histological analysis. Microarray analysis revealed that SCI led to the up-regulation of OPN and cathepsin D expression at 7 days and also of MT-1, MT-2, and tenascin-C expression at 1 day. Tenascin-C was re-up-regulated at 7 days. These values agreed with those of real-time RT-PCR analysis. By double labeling with in situ hybridization and fluorescence immunohistochemistry, MT-1, MT-2 and tenascin-C expression was observed in neurons and glial cells at 1 day, whereas at 7 days the main MT-2 and tenascin-C expression was found in fibronectin-positive fibroblasts. The main cathepsin D and OPN expression was observed in activated macrophages/microglia at 3 and 7 days. The five genes picked up by microarray gene expression profiling were shown to exhibit temporal and spatial changes of expression after SCI. This system is potentially useful for identifying genes that are involved in the response to SCI. [ABSTRACT FROM AUTHOR]

The minimization of a genome is necessary to identify experimentally the minimal gene set that contains only those genes that are essential and sufficient to sustain a functioning cell. Recent developments in genetic techniques have made it possible to generate bacteria with a markedly reduced genome. We developed a simple system for formation of markerless chromosomal deletions, and constructed and characterized a series of large-scale chromosomal deletion mutants ofEscherichia colithat lack between 2.4 and 29.7% of the parental chromosome. Combining deletion mutations changes cell length and width, and the mutant cells with larger deletions were even longer and wider than the parental cells. The nucleoid organization of the mutants is also changed: the nucleoids occur as multiple small nucleoids and are localized peripherally near the envelope. Inhibition of translation causes them to condense into one or two packed nucleoids, suggesting that the coupling of transcription and translation of membrane proteins peripherally localizes chromosomes. Because these phenotypes are similar to those of spherical cells, those may be a consequence of the morphological change. Based on the nucleoid localization observed with these mutants, we discuss the cellular nucleoid dynamics. [ABSTRACT FROM AUTHOR]

HAuCl4 was reduced by ethylene glycol, in the presence of polyvinylpyrrolidone (PVP) under microwave (MW) heating in a continuous wave (CW) mode for 2 min. Dominant products were polygonal nanoplates and close-to-spherical nanoparticles of gold. In addition, small amounts of single crystalline gold nanorods and nanowires (0.5–3% of total number of products) with diameters of 20–100 nm and lengths of 0.6–5 μm were produced. The diameter and length of gold nanorods and nanowires could be controlled by changing the HAuCl4·4H2O/PVP ratio. The formation mechanism of anisotropic gold nanostructures was discussed. [Copyright &y& Elsevier]

Semaphorin 3A (Sema3A) is a secreted repulsive axon guidance protein. It appears to play important roles in axon fasciculation, branching, neuronal migration, and tissue differentiation during embryonic development. In adults, Sema3A is expressed in spinal motoneurons and in some neurons in the brain. Here, we demonstrate changes in Sema3A expression in the spinal cord after complete transection and in the brain after spinal cord hemisection at the Th8 level in laboratory rats. Semi-quantitative reverse transcriptase-PCR analysis showed that the expression of Sema3A mRNA, which was present in the normal spinal cord, rapidly decreased after transection, reaching its lowest level 1 day after injury. Thereafter, Sema3A expression levels recovered and reached four-fifths of the normal level at 28 days. Double staining by in situ hybridization and fluorescence immunohistochemistry showed that Sema3A was expressed in NeuN-positive neurons, but not in glia in the spinal cord. Sema3A expression was up-regulated in the contralateral cerebral cortex and in the ipsilateral spinal trigeminal nucleus 1–3 days after spinal cord hemisection. It is likely that the up-regulation occurred in neurons whose descending fibers were transected. These results suggest that Sema3A is regulated differently in spinal motoneurons and brain neurons following axonal injury. [ABSTRACT FROM AUTHOR]

Osteopontin (OPN) is a secretory adhesive glycoprotein that is expressed in various tissues and plays a role in inflammation and tissue repair. It has been suggested that OPN plays a role in inflammation and wound healing after spinal cord injury; however, the expression of OPN and its function in the spinal cord under normal conditions and following spinal motoneuron injury have not been well characterized. Here we examined the expression of OPN mRNA before and after spinal root avulsion. OPN mRNA was detected at a low level in the normal spinal cord in a Northern blot analysis, but dramatically increased following avulsion. In situ hybridization and immunohistochemical studies demonstrated that OPN was present only in a subset of spinal motoneurons before avulsion. After avulsion, the number of OPN-expressing motoneurons increased, although the total number of motoneurons was reduced. OPN expression also became apparent in activated microglia/macrophages and astrocytes. These data suggest that the upregulation of OPN after spinal root avulsion is involved in two events, the protection of neurons and the post-traumatic inflammatory response in microglia/macrophages and astrocytes. [ABSTRACT FROM AUTHOR]

Investigates the indispensability of the Escherichia coli carbonic anhydrases YadF and CynT in cell proliferation at a low carbon dioxide partial pressure. Importance of YadF in cell growth in the absence of CynT; Growth of mutant strains lacking both of the anhydrases at high carbon dioxide concentrations.

Examines the nucleotide sequence and genetic structure of a novel carbaryl hydrolase gene. Persistence of agricultural pesticides in soil; Cloning of carbaryl degenerative genes; Presence of multiple isozymes on yeasts and other eukaryotes.

Develops an expression system for C-terminal chitin-binding domain of chitinase A1 (ChBDchiA1) produced from Escherichia coli. Non-interference of soluble substrates with the binding of ChBDchiA1; Recognition of ChBDchiA1 of a structure in chitin; Exhibition of ChBDchiA1 of binding activity over a wide range of hydrogen-ion concentration.

• Cutamesine treatment up-regulates BDNF expression in the lumbar spinal cord in normal rats. • Cutamesine treatment increased the number of BDNF-positive cells in injured rat spinal cord. • Cutamesine treatment significantly increased the number of serotonin boutons in the injured spinal cord. • Cutamesine treatment alone did not promote significant locomotor recovery. Cutamesine, a sigma-1 receptor agonist, functions in both neuroprotection and neurite outgrowth. We assessed the therapeutic effects of cutamesine in a rodent spinal cord injury (SCI) model to demonstrate pre-clinical proof-of-concept. First of all, in order to determine optimal cutamesine dose, cutamesine was administered to normal rats and BDNF protein levels in the lumbar spinal cord were assessed by Western blot. Next, for the SCI model, spinal cords of adult female Sprague-Dawley rats were contused using an Infinite Horizon Impactor. Two weeks post-injury, rats were randomly assigned to receive daily subcutaneous injections of either cutamesine (3.0 mg/kg/day) or saline (as a control) for another two weeks. Immunohistochemistry for BDNF and 5-HT was assessed at four and twelve weeks post-injury in the lumbar spinal cord. Locomotor function was assessed weekly using the BBB locomotor scale until twelve weeks after SCI and CatWalk XT 10.5 gait analysis was conducted at twelve weeks after SCI. In normal rats, cutamesine treatment (3.0 mg/kg/day) significantly up-regulated BDNF expression in the lumbar spinal cord. In SCI rats, cutamesine treatment (3.0 mg/kg/day) significantly increased the fluorescence intensity of neuronal BDNF and serotonin boutons in the injured spinal cord compared to saline. However, cutamesine treatment did not promote significant locomotor recovery. Recent work indicates that cutamesine treatment alone did not promote locomotor recovery in spite of immunohistological changes. Future work will explore the influence of combining cutamesine with other treatment promoting plasticity (e.g. rehabilitative training) in SCI rats. [ABSTRACT FROM AUTHOR]

Vaccines that efficiently target severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the etiological agent for coronavirus disease (COVID-19), are the best means for controlling viral spread. This study evaluated the efficacy of the COVID-19 vaccine S-268019-b, which comprises the recombinant full-length SARS-CoV-2 spike protein S-910823 (antigen) and A-910823 (adjuvant). In addition to eliciting both Th1-type and Th2-type cellular immune responses, two doses of S-910823 plus A-910823 induced anti-spike protein IgG antibodies and neutralizing antibodies against SARS-CoV-2. In a SARS-CoV-2 challenge test, S-910823 plus A-910823 mitigated SARS-CoV-2 infection-induced weight loss and death and inhibited viral replication in mouse lungs. S-910823 plus A-910823 promoted cytokine and chemokine at the injection site and immune cell accumulation in the draining lymph nodes. This led to the formation of germinal centers and the induction of memory B cells, antibody-secreting cells, and memory T cells. These findings provide fundamental property of S-268019-b, especially importance of A-910823 to elicit humoral and cellular immune responses. [ABSTRACT FROM AUTHOR]

Purpose: To assess the usefulness of software analysis using dynamic-ventilation CT for localized pleural adhesion (LPA).Materials and Methods: Fifty-one patients scheduled to undergo surgery underwent both dynamic-ventilation CT and static chest CT as preoperative assessments. Five observers independently evaluated the presence and severity of LPA on a three-point scale (non, mild, and severe LPA) for 9 pleural regions (upper, middle, and lower pleural aspects on ventral, lateral, and dorsal areas) on the chest CT by three different methods by observing images from: static high-resolution CT (static image); dynamic-ventilation CT (movie image), and dynamic-ventilation CT while referring to the adhesion map (movie image with color map), which was created using research software to visualize movement differences between the lung surface and chest wall. The presence and severity of LPA was confirmed by intraoperative thoracoscopic findings. Parameters of diagnostic accuracy for LPA presence and severity were assessed among the three methods using Wilcoxon signed rank test in total and for each of the three pleural aspects.Results: Mild and severe LPA were confirmed in 14 and 8 patients. Movie image with color map had higher sensitivity (56.9 ± 10.7 %) and negative predictive value (NPV) (91.4 ± 1.7 %) in LPA detection than both movie image and static image. Additionally, for severe LPA, detection sensitivity was the highest with movie image with color map (82.5 ± 6.1 %), followed by movie image (58.8 ± 17.0 %) and static image (38.8 ± 13.9 %). For LPA severity, movie image with color map was similar to movie image and superior to static image in accuracy as well as underestimation and overestimation, with a mean value of 80.2 %.Conclusion: Software-assisted dynamic-ventilation CT may be a useful novel imaging approach to improve the detection performance of LPA. [ABSTRACT FROM AUTHOR]

Attribute-Based Group Signature (ABGS) schemes permit any group member with required attributes to generate signatures for the sake of the group anonymously. Even though existing ABGS schemes with Verifier-local Revocation (VLR) method facilitate efficient user and attribute revocation, they cannot achieve stronger security for the users and user attributes. In this paper, we present a new approach to overcome this weakness delivering a new ABGS scheme with VLR that achieves stronger security, almost full anonymity, for both users and their attributes. We construct our scheme from lattices as lattice cryptography is quantum resist. Moreover, we present a simple member joining protocol and a new zero-knowledge argument system that supports the new scheme. Finally, we prove that the proposed scheme meets the security requirements of almost full anonymity, traceability, and non-frameability. [ABSTRACT FROM AUTHOR]

Background: Localized pleural adhesion (LPA) evaluation in the apical region is difficult even with four-dimensional ultra-low-dose computed tomography (4D-ULDCT) in the supine position due to smaller pleural movements.Purpose: To assess usability of 4D-ULDCT in the lateral decubitus (LD) position for LPA detection in the apical region.Material and Methods: Forty-seven patients underwent 4D-ULDCT of a single respiration cycle with 16-cm coverage of body axis in supine and LD positions with the affected lung uppermost. Intraoperative thoracoscopic findings confirmed LPA presence. A pleural point and a corresponding point on costal outer edge were placed in identical axial planes at end-inspiration. Pleuro-chest wall distance between two points (PCD) was calculated at each respiratory phase. In the affected lung, average change in amount of PCD (PCDACA) was compared between patients with and without LPA in total and two sub-groups (non-COPD and COPD, non-emphysematous and emphysematous patients) in supine and non-dependent (ND) LD positions. Receiver operating characteristic (ROC) curve analysis was performed to determine optimal thresholds in PCDACA for differentiating patients with LPA from those without.Results: In COPD/emphysematous patients and total population, PCDACA with LPA was smaller than in those without in the supine and NDLD positions for overall, lateral, and dorsal regions. For the lateral region in COPD patients, area under ROC curve (AUC) increased from supine (0.64) to NDLD position (0.81). For the dorsal region in emphysematous patients, AUC increased from supine (0.76) to NDLD position (0.96).Conclusion: 4D-ULDCT in LD position may be useful for LPA detection in apical regions for COPD and/or emphysematous patients. [ABSTRACT FROM AUTHOR]

Background: Extraintestinal pathogenic E. coli (ExPEC) is a common gram-negative organism causing various infections, including urinary tract infections (UTIs), bacteremia, and neonatal meningitis. The cjrABC-senB gene cluster of E. coli contributes to ExPEC virulence in the mouse model of UTIs. Consistently, the distribution of cjrABC-senB is epidemiologically associated with human UTIs caused by E. coli. cjrABC-senB, which has previously been proposed to encode an iron uptake system, may facilitate ExPEC survival in the iron availability-restricted urinary tract. Given that the bloodstream is also an iron limited environment to invading bacteria, the pathogenic role of cjrABC-senB in ExPEC bacteremia, however, remains to be investigated. Methods: The ability of ExPEC RS218 strains with and without cjrABC-senB to survive in the mouse bloodstream and human serum was evaluated. Subsequently, the role of this gene cluster in the ExPEC interaction with the complement system was evaluated. Finally, the distribution of cjrABC-senB in human clinical E. coli isolates was determined by PCR. The frequency of cjrABC-senB in bacteremia isolates that were not associated with UTIs (non-UTI bacteremia isolates) was compared with that in UTI-associated isolates and fecal isolates. Results: Expression of cjrABC-senB attenuated the survival of RS218 in the mouse bloodstream and human serum. The cjrABC-senB-harboring strains triggered enhanced classical- and alternative-complement pathway activation and became more vulnerable to complement-mediated killing in serum. cjrA was identified as the major gene responsible for the attenuated serum survival. Expressing cjrABC-senB and cjrA increased bacterial susceptibility to detergent and induced periplasmic protein leakage, suggesting that the expression of these genes compromises the integrity of the outer membrane of ExPEC. In addition, the frequency of cjrABC-senB in non-UTI bacteremia isolates was significantly lower than that in UTI-associated isolates, while the frequencies in non-UTI bacteremia isolates and fecal isolates showed no significant difference. Consistently, this epidemiological investigation suggests that cjrABC-senB does not contribute to E. coli bacteremia in humans. Conclusion: The contribution of cjrABC-senB to the pathogenesis of ExPEC is niche dependent and contradictory because the genes facilitate ExPEC UTIs but hinder bacteremia. The contradictory niche-dependent characteristic may benefit the development of novel strategies against E. coli-caused infections. [ABSTRACT FROM AUTHOR]

Background: Extraintestinal pathogenic E. coli (ExPEC) remains one of the most prevalent bacterial pathogens that cause extraintestinal infections, including neonatal meningitis, septicemia, and urinary tract (UT) infections (UTIs). Antibiotic therapy has been the conventional treatment for such infections, but its efficacy has decreased due to the emergence of antibiotic-resistant bacteria. Identification and characterization of bacterial factors that contribute to the severity of infection would facilitate the development of novel therapeutic strategies. The ExPEC periplasmic protease Prc contributes to the pathogen's ability to evade complement-mediated killing in the serum. Here, we further investigated the role of the Prc protease in ExPEC-induced UTIs and the underlying mechanism. Methods: The uropathogenic role of Prc was determined in a mouse model of UTIs. Using global quantitative proteomic analyses, we revealed that the expression of FliC and other outer membrane-associated proteins was altered by Prc deficiency. Comparative transcriptome analyses identified that Prc deficiency affected expression of the flagellar regulon and genes that are regulated by five extracytoplasmic signaling systems. Results: A mutant ExPEC with a prc deletion was attenuated in bladder and kidney colonization. Global quantitative proteomic analyses of the prc mutant and wild-type ExPEC strains revealed significantly reduced flagellum expression in the absence of Prc, consequently impairing bacterial motility. The prc deletion triggered downregulation of the flhDC operon encoding the master transcriptional regulator of flagellum biogenesis. Overexpressing flhDC restored the prc mutant's motility and ability to colonize the UT, suggesting that the impaired motility is responsible for attenuated UT colonization of the mutant. Further comparative transcriptome analyses revealed that Prc deficiency activated the σE and RcsCDB signaling pathways. These pathways were responsible for the diminished flhDC expression. Finally, the activation of the RcsCDB system was attributed to the intracellular accumulation of a known Prc substrate Spr in the prc mutant. Spr is a peptidoglycan hydrolase and its accumulation destabilizes the bacterial envelope. Conclusions: We demonstrated for the first time that Prc is essential for full ExPEC virulence in UTIs. Our results collectively support the idea that Prc is essential for bacterial envelope integrity, thus explaining how Prc deficiency results in an attenuated ExPEC. [ABSTRACT FROM AUTHOR]

Escherichia coli 16S, 23S, and 5S ribosomal RNAs (rRNAs) are transcribed as a single primary transcript, which is subsequently processed into mature rRNAs by several RNases. Three RNases (RNase III, RNase E, and RNase G) were reported to function in processing the 5′-leader of precursor 16S rRNA (pre-16S rRNA). Previously, we showed that a novel essential YqgF is involved in that processing. Here we investigated the ribosome subunits of the yqgFts mutant by LC-MS/MS. The mutant ribosome had decreased copy numbers of ribosome protein S1, suggesting that the yqgF gene enables incorporation of ribosomal protein S1 into ribosome by processing of the 5′-end of pre-16S rRNA. The ribosome protein S1 is essential for translation in E. coli; therefore, our results suggest that YqgF converts the inactive form of newly synthesized ribosome into the active form at the final step of ribosome assembly. [ABSTRACT FROM AUTHOR]

The minimal set of genetic information necessary and sufficient to sustain a functioning cell contains not only trans-acting genes, but also cis-acting chromosomal regions that cannot be complemented by plasmids carrying these regions. In Escherichia coli (E. coli), only one chromosomal region, the origin of replication has been identified to be cis-acting. We constructed a series of mutants with long-range deletions, and the chromosomal regions containing trans-acting essential genes were deleted in the presence of plasmids complementing the deleted genes. The deleted regions cover all regions of the chromosome except for the origin and terminus of replication. The terminus affects cell growth, but is not essential. Our results indicate that the origin of DNA replication is the only vital, unique cis-acting DNA sequence in the E. coli chromosome necessary for survival. [ABSTRACT FROM AUTHOR]

The Escherichia coli hda gene codes for a DnaA-related protein that is essential for the regulatory inactivation of DnaA (RIDA), a system that controls the initiation of chromosomal replication. We have identified the ygfZ gene, which encodes a folate-binding protein, as a suppressor of hda mutations. The ygfZ null mutation suppresses an hda null mutation. The over-initiation and abortive elongation phenotypes conferred by the hda mutations are partially suppressed in an hda ygfZ background. The accumulation of the active form of DnaA, ATP-DnaA, in the hda mutant is suppressed by the disruption of the ygfZ gene, indicating that YgfZ is involved in regulating the level of ATP-DnaA. Although ygfZ is not an essential gene, the ygfZ disruptant grows slowly, especially at low temperature, demonstrating that this gene is important for cellular proliferation. We have identified mnmE ( trmE) as a suppressor of the ygfZ disruption. This gene encodes a GTPase involved in tRNA modification. Examination of RNA modification in the ygfZ mutant reveals reduced levels of 2-methylthio N 6-isopentenyladenosine, indicating that YgfZ participates in the methylthio-group formation of this modified nucleoside in some tRNAs. These results suggest that YgfZ is a key factor in regulatory networks that act via tRNA modification. [ABSTRACT FROM AUTHOR]

Biogenesis of ribosomes is a complex process mediated by many factors. While its transcription proceeds, ribosomal RNA (rRNA) folds itself into a characteristic three-dimensional structure through interaction with ribosomal proteins, during which its ends are processed. Here, we show that the essential protein YqgF, a RuvC family protein with an RNase-H-like motif, is involved in the processing of pre-16S rRNA during ribosome maturation. Indeed, pre-16S rRNA accumulated in cells of a temperature-sensitive yqgF mutant ( yqgF ts ) cultured at a non-permissive temperature. In addition, purified YqgF was shown to process the 5′ end of pre-16S rRNA within 70S ribosomes in vitro . Mass spectrometry analysis of the total proteins in the yqgF ts mutant cells showed that the expression of genes containing multiple Shine–Dalgarno-like sequences was observed to be lower than in wild type. These results are interpreted to indicate that YqgF is involved in a novel enzymic activity necessary for the processing of pre-16S rRNA, thereby affecting elongation of translation. [ABSTRACT FROM AUTHOR]

Macrophage migration inhibitory factor (MIF) is a multipotential protein that acts as a proinflammatory cytokine, a pituitary hormone, and a cell proliferation and migration factor. The objective of this study was to elucidate the role of MIF in spinal cord injury (SCI) using female MIF knockout (KO) mice. Mouse spinal cord compression injury was produced by application of a static load (T8 level, 20 g, 5 min). We analyzed the motor function of the hind limbs and performed histological examinations. Hind-limb function recovered significantly in the KO mice starting from three weeks after injury. Cresyl-violet staining revealed that the number of surviving neurons in the KO mice was significantly larger than that of WT mice six weeks after injury. Immunohistochemical analysis revealed that the number of NeuN/caspase-3-active, double-positive, apoptotic neurons in the KO mice was significantly smaller than that of the WT mice 24 and 72 h after SCI. These results were related to in-vitro studies showing increased resistance of cerebellar granular neurons from MIF-KO animals to glutamate neurotoxicity. These results suggest that MIF existence hinders neuronal survival after SCI. Suppression of MIF may attenuate detrimental secondary molecular responses of the injured spinal cord. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the efficacy in adult rat completely transected spinal cord of adenovirus vector-mediated brain-derived neurotrophic factor (BDNF) ex vivo gene transfer to bone marrow stromal cells (BMSC). BMSC were infected with adenovirus vectors carrying β-galactosidase (AxCALacZ) or BDNF (AxCABDNF) genes. The T8 segment of spinal cord was removed and replaced by graft containing Matrigel alone (MG group) or Matrigel and BMSC infected by AxCALacZ (BMSC-LacZ group) or AxCABDNF (BMSC-BDNF group). Axons in the graft were evaluated by immunohistochemistry and functional recovery was assessed with BBB locomotor scale. In the BMSC-BDNF group, the number of fibers positive for growth associated protein-43, tyrosine hydroxylase, and calcitonin gene-related peptide was significantly larger than numbers found for the MG and BMSC-LacZ groups. Rats from BMSC-BDNF and BMSC-LacZ groups showed significant recovery of hind limb function compared with MG rats; however, there was no significant difference between groups in degree of functional recovery. These findings demonstrate that adenovirus vector-mediated ex vivo gene transfer of BDNF enhances the capacity of BMSC to promote axonal regeneration in this completely transected spinal cord model; however, BDNF failed to enhance hind limb functional recovery. Further investigation is needed to establish an optimal combination of cell therapy and neurotrophin gene transfer for cases of spinal cord injury. [ABSTRACT FROM AUTHOR]

Abstract: Shape and size controlled synthesis of gold nanocrystals have been studied by using a microwave (MW)-polyol method. When Au3+ in HAuCl4·4H2O was reduced in ethylene glycol (EG) in the presence of poly(vinylpyrrolidone) (PVP) at a low HAuCl4 concentration, mixtures of spherical, triangular, octahedral, and decahedral particles were prepared in a few minutes. It was found that the shape selective oxidative etching by AuCl4 − +Cl− anions and crystal growth occurred simultaneously at high HAuCl4 concentrations. The addition of MCl (M=Na, K, H) was very effective to enhance the shape selective oxidative etching by AuCl4 − +Cl− anions. With increasing the AuCl4 − and Cl− concentrations, yields of large plate-type and ribbon-type single twin particles increased, while those of spherical and octahedral particles decreased in most cases. Possible etching and growth mechanisms are discussed from dramatic color and absorption spectral changes of Au seeds in the HAuCl4·4H2O/PVP/EG solution with increasing the solution temperature and from TEM images before and after oxidative etching. The present results provide a new simple technique for the rapid shape and size controlled synthesis of Au nanocrystals under MW heating. [Copyright &y& Elsevier]

Macrophage migration inhibitory factor (MIF) is a multipotential protein that acts as a pro-inflammatory cytokine, pituitary hormone, immunoregulator, and mitogen. To elucidate function of MIF in spinal cord injury, we examined expression of MIF after compression-induced spinal cord injury using Northern blot analysis, in situ hybridization and immunohistochemistry. The MIF mRNA was up-regulated in injured spinal cord, peaking 3 days after injury shown by Northern blot analysis. In situ hybridization revealed up-regulation of MIF in microglia accumulating in the lesion epicenter 3 days after injury and astrocytes around the cystic cavity 1 week after injury. Double staining showed co-localization of MIF and tomato lectin in the lesioned site, indicating that microglia accumulating to the lesion epicenter express MIF. The time course of MIF expression is different from that of previous reports about cytokine expression peaking at earlier time points; thus, it is unlikely that MIF acts as a pro-inflammatory factor in the present study. The MIF may contribute to proliferation of astrocytes around the lesioned site in spinal cord injury because of its cell proliferation-promoting property. [ABSTRACT FROM AUTHOR]

In advanced lung cancer treatment, immunotherapy provides durable responses in some patients. However, other patients experience progressive disease and the resistance mechanisms to immunotherapy have yet been fully elucidated. Small cell transformation of non‐small cell lung cancer (NSCLC) is commonly recognized as one of the resistance mechanisms to epidermal growth factor receptor (EGFR)‐tyrosine kinase inhibitors in EGFR‐mutant NSCLC treatment. As a resistant mechanism for immunotherapy, we report the first case of small cell transformation in 2017. Since then, eight similar cases have been reported and the concept of small cell transformation is now becoming more prevalent as a mechanism of immunotherapy resistance. In our facility, we have experienced four cases of small cell transformation after immunotherapy (including the reported case in 2017). The histology of each primary tumor was squamous cell carcinoma, large cell type neuroendocrine carcinoma, or poorly differentiated NSCLC. None had driver gene mutations. Nivolumab was administered in all four cases and atezolizumab was administered as a next line to nivolumab treatment in one case. The best response to immunotherapy was partial response or stable disease. There was a wide range of periods from the start of immunotherapy to confirmation of small cell transformation (from 2 weeks to almost 3 years). In conclusion, small cell transformation is an important resistance mechanism in cancer immunotherapy. When NSCLC progresses after immunotherapy, the possibility of small cell transformation and rebiopsy should always be encouraged, as it leads to clarification of the resistance mechanisms and frequency. [ABSTRACT FROM AUTHOR]

Background: A synergistic effect of cyclooxygenase inhibitors (COX‐I) and immune checkpoint inhibitors (ICIs) has been suggested. However, the impact of COX‐I on the efficacy of ICIs is unclear. Here, we aimed to evaluate the relationship between COX‐I use and the efficacy of ICI in patients with non‐small cell lung cancer (NSCLC). Methods: We retrospectively reviewed NSCLC patients who received ICI monotherapy. We defined COX‐I use as regular use of COX‐I other than low‐dose aspirin during the initiation of ICIs to the first evaluation of efficacy. The efficacy of ICIs was evaluated with response rate (RR), disease control rate (DCR), progression free survival (PFS), and overall survival (OS). Differences in baseline characteristics by COX‐I use were controlled by using an inverse probability of treatment weighting (IPW) adjusted analysis. Results: A total of 198 patients with NSCLC received ICIs; 128, 50, and 20 patients received nivolumab, pembrolizumab, and atezolizumab, respectively; there were 65 (32.8%) COX‐I users. While there was no significant difference in RR (15.4% vs. 13.5%; p = 0.828), DCR (41.5% vs. 49.6%; p = 0.294), PFS (median, 2.69 vs. 3.68 months; 95% confidence intervals [CI], 1.77–5.19 vs. 2.20–4.60 months; p = 0.630), COX‐I users had significantly shorter OS than non‐COX‐I users (median, 6.08 vs. 16.10 months; 95% CI: 3.78–11.66 vs. 9.49–19.68 months; p = 0.003). On IPW adjusted analysis, there was no significant difference in OS (median, 7.85 vs. 15.11 months; 95% CI: 5.03–14.92 vs. 9.49–19.32 months; p = 0.081). Conclusions: There was no additional or negative impact of COX‐I use on the efficacy of ICIs in NSCLC. [ABSTRACT FROM AUTHOR]

Background: Over the past two decades, antiresorptive agent‐related osteonecrosis of the jaw (ARONJ) has become a growing concern. We examined the incidence of ARONJ and identified its risk factors in lung cancer patients in the real‐world clinical setting. To our knowledge, we are the first to do so. Patients and methods: We retrospectively analyzed lung cancer patients with bone metastases who had received anti‐resorptive agents (zoledronate or denosumab) at the National Hospital Organization Kyoto Medical Center from October 2012 to September 2018. All ARONJ cases were diagnosed by the dentists according to the established diagnostic criteria. Results: A total of 171 patients were reviewed, 13 (7.6%) of whom experienced ARONJ. Among the 13 patients, six (46.2%), four (30.8%) and three (23.1%) had adenocarcinoma, squamous carcinoma and not otherwise specified, respectively. ARONJ was stage 2 in three (23.1%) patients and stage 3 in 10 (76.9%). More cycles of antiresorptive agents (odds ratio [OR] = 11.54; 95% confidence interval [CI], 2.47–53.99; P < 0.01), use of immune checkpoint inhibitors (ICIs; OR = 5.05; 95% CI, 1.56–16.37; P < 0.01) and longer survival duration (≥2 years; OR = 12.16; 95% CI, 3.17–46.65; P < 0.01) were independently associated with ARONJ in a multivariate analysis. Conclusions: The incidence of ARONJ was relatively high in lung cancer patients with bone metastases. When using antiresorptive agents, oncologists should closely monitor patients for ARONJ during the course of treatment and regularly consult with dentists, especially in patients receiving ICIs. [ABSTRACT FROM AUTHOR]

Background: This study evaluated the efficacy and safety of retreatment with anti‐programmed death 1 (anti‐PD‐1) antibodies in patients with advanced non‐small cell lung cancer (NSCLC) after prior treatment with anti‐programmed death‐ligand 1 (anti‐PD‐L1) antibodies. Methods: Data (N = 15) on patients' characteristics, number of cycles, regimens, their best response and immune‐related adverse events (irAEs) were recorded retrospectively. Results: NSCLC was initially treated with anti‐PD‐L1 antibody atezolizumab (N = 14) or durvalumab (N = 1). No patients had a high (≥50%) tumor expression of PD‐L1. The median cycles for atezolizumab were five (range 1–15), and median progression‐free survival was 2.8 and 6.0 months for atezolizumab and durvalumab, respectively. Five (33.3%) and nine (60.0%) patients showed stable and progressive disease as their best response, respectively. No differences in irAEs between anti‐PD‐L1 and anti‐PD‐1 antibodies occurred. Conclusion: Patients treated with anti‐PD‐L1 antibodies for NSCLC received limited benefits from retreatment with anti‐PD‐1 antibodies. [ABSTRACT FROM AUTHOR]

Although it ameliorates lung cancer, immunotherapy with immune checkpoint inhibitors (ICIs) presents complications of infectious diseases, including tuberculosis. Incidence of tuberculosis during immunotherapy remains unclear. We found that 1.7% of patients developed active tuberculosis during immunotherapy at our institution. In patients with a positive interferon-gamma release assay status before ICI therapy, physicians should pay close attention to developing tuberculosis. [ABSTRACT FROM AUTHOR]