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Abstract Trichophyton rubrum is one of the most frequently isolated fungi in patients with dermatophytosis. Despite its clinical significance, the molecular mechanisms of drug resistance and pathogenicity of T. rubrum remain to be elucidated because of the lack of genetic tools, such as efficient gene targeting systems. In this study, we generated a T. rubrum strain that lacks the nonhomologous end-joining-related gene ku80 (Δku80) and then developed a highly efficient genetic recombination system with gene targeting efficiency that was 46 times higher than that using the wild-type strain. Cyp51A and Cyp51B are 14-α-lanosterol demethylase isozymes in T. rubrum that promote ergosterol biosynthesis and are the targets of azole antifungal drugs. The expression of cyp51A mRNA was induced by the addition of the azole antifungal drug efinaconazole, whereas no such induction was detected for cyp51B, suggesting that Cyp51A functions as an azole-responsive Cyp51 isozyme. To explore the contribution of Cyp51A to susceptibility to azole drugs, the neomycin phosphotransferase (nptII) gene cassette was inserted into the cyp51A 3′-untranslated region of Δku80 to destabilize the mRNA of cyp51A. In this mutant, the induction of cyp51A mRNA expression by efinaconazole was diminished. The minimum inhibitory concentration for several azole drugs of this strain was reduced, suggesting that dermatophyte Cyp51A contributes to the tolerance for azole drugs. These findings suggest that an efficient gene targeting system using Δku80 in T. rubrum is applicable for analyzing genes encoding drug targets.

Summary: The development of antifungal drugs requires novel molecular targets due to limited treatment options and drug resistance. Through chemical screening and establishment of a novel genetic technique to repress gene expression in Trichophyton rubrum, the primary causal fungus of dermatophytosis, we demonstrated that fungal Cdc42 and Rac GTPases are promising antifungal drug targets. Chemical inhibitors of these GTPases impair hyphal formation, which is crucial for growth and virulence in T. rubrum. Conditional repression of Cdc24, a guanine nucleotide exchange factor for Cdc42 and Rac, led to hyphal growth defects, abnormal cell morphology, and cell death. EHop-016 inhibited the promotion of the guanine nucleotide exchange reaction in Cdc42 and Rac by Cdc24 as well as germination and growth on the nail fragments of T. rubrum and improved animal survival in an invertebrate infection model of T. rubrum. Our results provide a novel antifungal therapeutic target and a potential lead compound.

Hydrogen gas (H2) has attracted attention as a next-generation clean energy source. Its efficient and safe preparation and utilization are crucial in both the industry and organic chemistry research. In this study, a Pt/CB (platinum on carbon bead)-catalyzed MW-mediated continuous-flow hydrogenation reaction was developed using methylcyclohexane (MCH) as the reducing agent (hydrogen carrier). Alkynes, alkenes, nitro groups, benzyl esters, and aromatic chlorides were chemoselectively hydrogenated using Pt/CB under MW-assisted continuous-flow conditions. This methodology represents a safe and energy-efficient hydrogenation process, as it eliminates the need for an external hydrogen gas supply or heating jackets as a heating medium. The further application of MW-mediated continuous-flow hydrogenation reactions is a viable option for the efficient generation and utilization of sustainable energy.

Abstract The synthesis of polycyclic aromatic compounds generally requires stoichiometric oxidants or homogeneous metal catalysts, however, the risk of contamination of inorganic residues can affect their properties. Here we present a microwave (MW)-assisted platinum on beaded activated carbon (Pt/CB)-catalyzed C–C bond formation of diarylacetylenes and aromatic hydrocarbons under continuous-flow conditions. Various fused aromatic compounds were continuously synthesized via dehydrogenative C(sp2)–C(sp2) and C(sp2)–C(sp3) bond formation with yields of up to 87% without the use of oxidants and bases. An activated, local reaction site on Pt/CB in the flow reaction channel reaching temperatures of more than three hundred degrees Celsius was generated in the catalyst cartridge by selective microwave absorption in CB with an absorption efficiency of > 90%. Mechanistic experiments of the transformation reaction indicated that a constant hydrogen gas supply was essential for activating Pt. This is an ideal reaction with minimal input energy and no waste production.

ABSTRACT Dermatophytes invade and colonize host superficial tissues via hyphal growth. Although cytoskeletal reorganization and its regulation are essential for hyphal growth, the molecular mechanisms in dermatophytes and their applicability as antifungal drug targets remain poorly understood. The p21-activated kinase (PAK) is a downstream effector of the small GTPases Rac and CDC42, also known as p21, and is involved in various molecular and cellular functions, including actin polymerization and cell morphogenesis. In this study, we investigated the contribution of the PAK protein TrCla4 to morphogenesis and mycelial growth in Trichophyton rubrum, the most frequently isolated fungus in dermatophytosis (athlete’s foot). The actin polymerization inhibitor, cytochalasin A inhibited actin accumulation at the hyphal tip and mycelial growth of T. rubrum, suggesting the involvement of the actin cytoskeleton in mycelial growth. In the Trcla4 knockout strain (ΔTrcla4), we observed defects in mycelial growth, hyphal branching, and the accumulation of polymerized actin at the hyphal tip. Chemical inhibitors of TrRac-dependent TrCla4 kinase activity, FRAX486 and IPA-3, also inhibited spore germination and mycelial growth. Interestingly, ΔTrcla4 showed no additional inhibition of mycelial growth when treated with these inhibitors, indicating that their inhibitory effects are primarily mediated through TrCla4. In an invertebrate dermatophyte infection model, animals infected with ΔTrcla4 had higher survival rates than those infected with the wild-type, and IPA-3 and FRAX486 treatments both significantly improved animal survival rates. These results suggest that the dermatophyte PAK promotes mycelial growth by facilitating actin polymerization at the hyphal tip, making it a potential therapeutic target for dermatophytosis. IMPORTANCE Superficial fungal infections, such as athlete’s foot, affect more than 10% of the world’s population and have a significant impact on quality of life. Despite the fact that treatment-resistant fungi are a concern, there are just a few antifungal drug targets accessible, as opposed to the wide range of therapeutic targets found in bacterial infections. As a result, additional alternatives are sought. In this study, we generated a PAK TrCla4 deletion strain (∆Trcla4) of Trichophyton rubrum. The ∆Trcla4 strain exhibited deficiencies in mycelial growth, hyphal morphology, and polarized actin localization at the hyphal tip. IPA-3 and FRAX486, small chemical inhibitors of mammalian PAK, were discovered to limit fungal mycelial proliferation. According to our findings, fungal PAKs are interesting therapeutic targets for the development of new antifungal medicines.

Abstract Trichosporon asahii is a conditional pathogenic fungus that causes severe and sometimes fatal infections in immunocompromised patients. While calcineurin, an essential component of a calcium-dependent signaling pathway, is known to regulate stress resistance and virulence of some pathogenic fungi, its role in T. asahii has not been investigated. Here, we demonstrated that calcineurin gene-deficient T. asahii mutants are sensitive to high temperature as well as cell-membrane and cell-wall stress, and exhibit decreased hyphal formation and virulence against silkworms. Growth of T. asahii mutants deficient in genes encoding subunits of calcineurin, cna1 and cnb1, was delayed at 40 °C. The cna1 and cnb1 gene-deficient mutants also showed sensitivity to sodium dodecyl sulfate, Congo red, dithiothreitol, and tunicamycin. On the other hand, these mutants exhibited no sensitivity to caffeine, sorbitol, monensin, CaCl2, LiCl, NaCl, amphotericin B, fluconazole, or voriconazole. The ratio of hyphal formation in the cna1 and cnb1 gene-deficient mutants was decreased. Moreover, the virulence of the cna1 and cnb1 gene-deficient mutants against silkworms was attenuated. These phenotypes were restored by re-introducing each respective gene into the gene-deficient mutants. Our findings suggest that calcineurin has a role in regulating the cellular stress response and virulence of T. asahii.

Abstract Trichosporon asahii is a pathogenic fungus that causes deep-seated fungal infections in immunocompromised patients. Established methods for generating gene-deficient T. asahii mutants exist, but the frequency of obtaining transformants by electroporation remains low. In the present study, we optimized the conditions for gene transfer by electroporation using a ku70 gene-deficient mutant with high recombination efficiency. Introducing a DNA fragment by electroporation into T. asahii cells on Sabouraud dextrose agar to generate a cnb1 gene-deficient mutant and incubating for 1 day led to the growth of approximately 100 transformants. When the incubation period was extended to 2 days or 5 days, however, only 2 or no transformants, respectively, were grown. The highest number of transformants was grown by electroporation when a square wave at 1.8 kV (9 kV/cm) was applied for 5 ms. In addition, the number of transformants increased with an increase in the length of the homologous region, and transformants did not grow when the homologous region was less than 500 base pairs. A DNA fragment was produced for deletion of the cnb1 gene by joint PCR, and the cnb1 gene-deficient mutant was obtained by introducing the DNA fragment by electroporation. These results indicate that DNA fragments produced by joint PCR can be used to generate gene-deficient mutants of T. asahii through gene transfer by electroporation. Graphical Abstract

Abstract Trichosporon asahii is a pathogenic fungus that causes severe, deep-seated fungal infections in neutropenic patients. Elucidating the infection mechanisms of T. asahii based on genetic studies requires a specific gene-targeting system. Here, we established an efficient gene-targeting system in a highly pathogenic T. asahii strain identified using the silkworm infection model. By comparing the pathogenicity of T. asahii clinical isolates in a silkworm infection model, T. asahii MPU129 was identified as a highly pathogenic strain. Using an Agrobacterium tumefaciens-mediated gene transfer system, we obtained a T. asahii MPU129 mutant lacking the ku70 gene, which encodes the Ku70 protein involved in the non-homologous end-joining repair of DNA double-strand breaks. The ku70 gene-deficient mutant showed higher gene-targeting efficiency than the wild-type strain for constructing a mutant lacking the cnb1 gene, which encodes the beta-subunit of calcineurin. The cnb1 gene-deficient mutant showed reduced pathogenicity against silkworms compared with the parental strain. These results suggest that an efficient gene-targeting system in a highly pathogenic T. asahii strain is a useful tool for elucidating the molecular mechanisms of T. asahii infection.

Abstract Background Thoracic ossification of ligamentum flavum (T-OLF), as one of the causes of thoracic myelopathy, is often combined with other spinal disorders. Concurrent lumbar spinal canal stenosis (LCS) is often obscured by symptoms due to T-OLF, leading to difficulty in identifying the origin of these neurological findings. It is common to be misdiagnosed or delayed diagnosis due to the complicated nature. We evaluated the prevalence, distribution, and clinical characteristics of OLF, especially in patients with LCS. Methods The authors performed a retrospective analysis of the outcomes of 61 patients who underwent thoracic surgeries performed for symptomatic T-OLF. In all the patients, whole spine lesions were evaluated preoperatively. We examined the factors related to poor outcomes (the recovery rate of the Japanese Orthopedic Association score for thoracic myelopathy is less than 40%) following OLF surgeries. We compared the clinical outcomes according to whether there was concurrent LCS, and determined the optimal surgical approach. Results The occurrence of T-OLF increased with age. Forty-six cases (75.4%) were considered to be tandem T-OLF and LCS (LCS group). An advanced age, and concurrent LCS were associated with a poor outcome after the surgery. The LCS group significantly included a greater number of elderly, and more light-weighted patients with Modic change in thoracic spine and a greater sagittal vertical axis, resulting in the lower neurological recovery. Additional lumbar surgery (13cases) effectively improved both the T-JOA and L-JOA scores (from 6.5 ± 2.0 points to 8.0 ± 1.8 points, p = 0.0406, and from 14.5 ± 4.7 points to 20.7 ± 2.6 points, p = 0.001, respectively) in OLF patients with LCS. Conclusions T-OLF was highly associated with other spinal disorders. Poor outcomes in T-OLF surgery could be associated with age and concurrent LCS, and an additional surgery for another lumbar lesion significantly improved neurological findings in T-OLF patients.

The environments of nursing home staff and residents have dramatically changed since the onset of the COVID-19 pandemic, with greater demand for infection control. This study aimed to clarify the changes and regional differences in the surrounding environment of nursing home residents as well as the working environment of staff, including oral health care, after the spread of SARS-CoV-2. A self-administered questionnaire survey was sent to nursing staff at about 40 nursing homes in different areas of Japan in September and October 2021. The questionnaire consisted of items centered around: (1) the surrounding environment of nursing home residents, (2) awareness and attitudes towards daily work among staff, and (3) attitudes to and procedures for oral health care among staff. A total of 929 respondents included 618 (66.5%) nursing care workers and 134 (14.4%) nurses. Regarding changes in resident daily life, 60% of staff perceived decreases in psychosocial and physical function after the start of the pandemic due to limited family communication and recreational activities, especially in urban areas. Concerning infection control, most respondents adopted routines of disinfecting hands before and after their duties. Oral health care was part of the regular duties of over 80% of respondents. Many participants answered that the frequency and time of oral health care only slightly changed after the onset of COVID-19, but many also reported disinfecting hands both before and after oral health care, particularly in rural areas. Our findings suggested that the COVID-19 pandemic decreased the daily living activities of residents, leading to psychosocial and physical decline, especially in urban areas. The results also indicated that the spread of SARS-CoV-2 triggered improvements in the awareness and attitudes towards infection control in daily work, including oral health care, among nursing care staff, notably in rural areas. Such an effect may contribute to a more positive perception of oral health care infection measures after the pandemic.

Introduction: Anterior decompression and fusion have shown favorable neurologic outcomes in patients with cervical myelopathy. However, implant migration sometimes occurs immediately after multilevel anterior cervical corpectomy with fusion (ACCF). Risk factors associated with early bone graft migration have not been precisely documented. The study aimed to investigate how frequently bone graft subsidence occurs after ACCF and to determine the factors affecting implant migration. Methods: Forty-seven consecutive patients who underwent ACCF for ossification of the posterior longitudinal ligament at our hospital between 2007 and 2015 and were able to complete 1 year of follow-up were enrolled. Patients treated with hybrid fixation were excluded. Data on demographics and radiographic findings, namely, fused segment angle and fused segment height (FSH), were collected. Implant migration was defined as subsidence of >3 mm. The patients were divided into 2-segment (2F), 3-segment (3F), and 4-segment (4F) groups. Results were compared between the groups using one-way analysis of variance, the Mann-Whitney U test, and the chi-square test. Results: Mean age was 61.6 years in the 2F group (n = 17), 62.1 years in the 3F group (n = 21), and 69 years in the 4F group (n = 9). There were no significant between-group differences in demographics or clinical characteristics. Implant subsidence occurred in 3 cases (17.6%) in the 2F group, 4 (19%) in the 3F group, and 3 (33.3%) in the 4F group. Revision surgery was required in 2 cases (1 patient each in the 3F and 4F groups). Logistic regression analysis showed a significant association of increased FSH and increased risk of postoperative implant subsidence. Conclusions: A postoperative increase in FSH may affect graft stability and lead to early implant migration.

Silicon carbide (SiC)-supported palladium (Pd) catalysts [3% Pd/SiC and a 3% Pd-diethylenetriamine (DETA)/SiC complex] for chemoselective hydrogenation under batch- and continuous-flow conditions were developed. The alkyne, alkene, azide, nitro, and benzyloxycarbonyl-protected aromatic amine (N-Cbz) functionalities were chemoselectively reduced in the presence of 3% Pd/SiC. By contrast, benzyl ether, alkyl N-Cbz, epoxide, aromatic chloride, aromatic ketone, and tert-butyldimethylsilyl ether were tolerant to the 3% Pd/SiC-catalyzed hydrogenation. The combined use of 3% Pd/SiC and DETA demonstrated excellent chemoselectivity toward the semihydrogenation of various mono- and disubstituted alkynes under batch- and continuous-flow conditions. Furthermore, compared with the separate use of 3% Pd/SiC and DETA, the newly developed 3% Pd(DETA)/SiC-packed in a cartridge showed higher chemoselectivity toward the continuous-flow semihydrogenation of alkyne over 24 h.

Abstract Background Atlantoaxial subluxation (AAS) is characterized by excessive movement at the junction between the atlas (C1) and axis (C2) as a result of either a bony or ligamentous abnormality. Surgical intervention is a therapeutic choice for AAS. In addition to C1 laminectomy (LAM), surgical fixation for subluxation or instability is performed by various techniques. While surgical treatment options for AAS have increased, the outcomes of different surgical techniques remain unclear. Methods The authors conducted a retrospective analysis of the outcomes of 30 consecutive spinal surgeries performed for AAS patients, C1 LAM in 11 cases and C1/2 fixation in 19 cases. We investigated the correlation between the clinical outcomes and the surgical methods. We also examined the factors related to poor outcomes (the recovery rate of the Japanese Orthopedic Association score for cervical myelopathy

Introduction: Patients treated with revision surgery after lumbar decompression with fusion typically have persistent low back pain and lower extremity numbness compared with patients treated with only primary surgery. No well-designed study has investigated the persistence and degree of pain after revision surgery following instrumented operation. The purpose of this study is to compare residual pain among patients who underwent reoperation and those who underwent only primary surgery for lumbar degenerative disorder using patient-based evaluation. Methods: We reviewed 350 consecutive patients (143 men, 207 women, mean age 63 years) treated with primary lumbar instrumented surgery between October 2010 and February 2014 at our institution and followed up for 2 years postoperatively. Patients were categorized into three groups based on number of levels fused: 1-segment, 2-segment, and 3-segment fusion (1F, 2F, and 3F groups, respectively). We used the Japanese Orthopedic Association Back Pain Evaluation Questionnaire (JOABPEQ) and visual analog scales (VASs) for low back pain and lower extremity pain to evaluate pain intensity pre- and postoperatively. Results: Salvage surgery for late-phase complications was required in 5 cases (2.4%), 6 cases (11.3%), and 11 cases (12.1%) in the 1F, 2F, and 3F groups, respectively. In the 1F and 2F groups, patients treated with revision surgery had unsatisfactory improvement in the pain domain of JOABPEQ and VASs for low back pain and lower extremity pain compared with patients with only primary short fusion surgery. The 3F group showed no significant differences between patients who underwent reoperation and those who underwent only primary surgery. Conclusion: Low back pain and lower extremity pain often persist after revision surgery in patients treated with short fusion (2-segment) operation. We need to follow pain states in such patients.

Abstract Background In patients with ossification of the posterior longitudinal ligament (OPLL) in the cervical spine, it is well known that the thoracic ossified lesions often coexist with the cervical lesions and can cause severe myelopathy. However, the prevalence of OPLL at each level of the thoracic and lumbar spinal segments is unknown. The aims of this study were to investigate how often OPLL occurs at each level in the thoracolumbar spine in patients with a radiological diagnosis of cervical OPLL and to identify the spinal levels most likely to develop ossification. Methods Data were collected from 20 institutions in Japan. Three hundred and twenty-two patients with a diagnosis of cervical OPLL were included. The OPLL index (OP index), defined as the sum of the vertebral body and intervertebral disc levels where OPLL is present, was used to determine disease severity. An OP index ≥20 was defined as severe OPLL. The prevalence of OPLL at each level of the thoracic and lumbar spinal segments was calculated. Results Women were more likely to have ossified lesions in the thoracolumbar spine than men. Severe OPLL was significantly more common in women than in men (20% vs. 4.5%). For thoracic vertebral OPLL, the most frequently affected was the T1 segment in both men and women, followed by the T1/2 and T3/4 intervertebral levels in men and women, respectively. Ossified lesions were frequently seen at the intervertebral and vertebral levels around the cervicothoracic and thoracolumbar junctions in men with severe OPLL, whereas OPLL was more diffusely distributed in the thoracic spine in women with severe OPLL. Conclusion Thoracolumbar OPLL occurred most often at T1 in men and at T3/4 in women. In severe OPLL cases, although ossified lesions were frequently seen at the intervertebral and vertebral levels around the cervicothoracic and thoracolumbar junctions in men, OPLL could be observed more diffusely in the thoracic spine in women.

Objectives: As the population ages, the number of lumbar spinal surgeries performed on sarcopenic patients will increase. The purpose of this study was to investigate the prevalence of sarcopenia and evaluated its impact on the results of lumbar spinal surgery. Methods: This study included 2 groups: One group consisted of patients who underwent whole-body dual-energy X-ray absorptiometry (DXA) scanning before the option of undergoing surgery for lumbar spinal disease (LSD group) and a second group consisted of patients underwent DXA scanning for osteoporosis screening under hospital watch at the geriatric medicine department (control group). In order to evaluate the impact of sarcopenia on the clinical outcome of lumbar spinal surgery, the Japanese Orthopedic Association (JOA) score, the recovery rate based on the JOA score, and visual analogue scale (VAS) scores for lower back pain, lower extremity pain, and lower extremity numbness were compared within the LSD group. Results: The prevalence of sarcopenia showed no statistical difference between groups (control group, 50.7%; LSD group, 46.5%). In the LSD group, while the changes in VAS scores showed no statistical difference between the nonsarcopenia subgroup and sarcopenia subgroup, the sarcopenia subgroup demonstrated inferior JOA scores and recovery rates at the final follow-up when compared with the nonsarcopenia subgroup (P

Abstract Background Concurrent cervical and lumbar spinal canal stenosis is known as tandem spinal stenosis (TSS). As research on TSS is limited, there is no consensus on the optimal surgical approach to this problem. We evaluated the prevalence and clinical characteristics of TSS in patients with symptomatic lumbar spinal canal stenosis (LCS). Methods The authors performed a retrospective analysis of the outcomes of 565 patients who underwent lumbar surgeries performed for symptomatic LCS. In all the patients, both the cervical and lumbar regions were evaluated preoperatively, and we compared TSS patients and non-TSS patients in terms of multiple clinical parameters. In the TSS patients, we investigated the ratio and clinical outcomes of additional cervical surgeries performed on TSS patients. Results Two hundred two cases (35.8%) were considered to be TSS. Twenty-eight patients (5.0%) underwent a cervical operation during the follow-up period. There were no differences between the radiographic TSS patients and non-TSS patients in terms of preoperative lumbar-Japanese Orthopedic Association (L-JOA) scores, postoperative L-JOA scores, and the L-JOA recovery rate (14.8 ± 4.4 points vs 14.2 ± 4.6 points, 23.9 ± 4.3 points vs 23.1 ± 4.5 points, 63.7 ± 28.2% vs 60.3 ± 27.9%, respectively), while the TSS group included a greater number of hypertension cases. The recovery rate L-JOA scores of patients who underwent additional cervical surgeries were significantly lower compared with the rate of patients who experienced treatment for only lumbar lesions (62.8 ± 25.8% vs 39.8 ± 35.5%, p = 0.0003). However, additional cervical surgery still improved both the cervical myelopathy-Japanese Orthopedic Association (C-JOA) and L-JOA scores in TSS patients with symptomatic cervical lesion (from 10.3 ± 2.8 points to 12.1 ± 3.0 points, p = 0.0302; from 14.8 ± 7.3 points to 19.9 ± 5.0 points, p = 0.0331, respectively). In these patients, there were no significant differences in the recovery rate of both C-JOA and L-JOA scores between the single-stage surgery group and the staged surgery group (40.7 ± 35.8% vs 20.7 ± 16.1%; 50.9 ± 25.1% vs 34.2 ± 39.3%, respectively). Conclusions Radiographic co-existing cervical stenosis did not affect surgical outcomes for LCS, although symptomatic cervical lesion affected neurological score after lumbar surgery. An additional surgery for cervical lesion significantly improved neurological findings in TSS patients.

Various studies have found a high incidence of early graft dislodgement after multilevel corpectomy. Although a hybrid fusion technique was developed to resolve implant failure, the hybrid and conventional techniques have not been clearly compared in terms of perioperative complications in patients with severe ossification of the posterior longitudinal ligament (OPLL) involving three or more levels. The purpose of this study was to compare clinical and radiologic outcomes between anterior cervical corpectomy with fusion (ACCF) and anterior hybrid fusion for the treatment of multilevel cervical OPLL. We therefore retrospectively reviewed the clinical and radiologic data of 53 consecutive patients who underwent anterior fusion to treat cervical OPLL: 30 underwent ACCF and 23 underwent anterior hybrid fusion. All patients completed 2 years of follow-ups. Implant migration was defined as subsidence > 3 mm. There were no significant differences in demographics or clinical characteristics between the ACCF and hybrid groups. Early implant failure occurred significantly more frequently in the ACCF group (5 cases, 16.7%) compared with the hybrid group (0 cases, 0%). The fusion rate was 80% in the ACCF group and 100% in the hybrid group. Although both procedures can achieve satisfactory neurologic outcomes for multilevel OPLL patients, hybrid fusion likely provides better biomechanical stability than the conventional ACCF technique.

Lateral lumbar interbody fusion (LLIF) is increasingly performed as corrective surgery for patients with adult spinal deformity (ASD). This paper compares the surgical results of LLIF and conventional posterior lumbar interbody fusion (PLIF)/transforaminal lumbar interbody fusion (TLIF) in ASD using a propensity score matching analysis. We retrospectively reviewed patients with ASD who received LLIF and PLIF/TLIF, and investigated patients’ backgrounds, radiographic parameters, and complications. The propensity scores were calculated from patients’ characteristics, including radiographic parameters and preoperative comorbidities, and one–to-one matching was performed. Propensity score matching produced 21 matched pairs of patients who underwent LLIF and PLIF/TLIF. All radiographic parameters significantly improved in both groups at the final follow-up compared with those of the preoperative period. The comparison between both groups demonstrated no significant difference in terms of postoperative pelvic tilt, lumbar lordosis (LL), or pelvic incidence–LL at the final follow-up. However, the sagittal vertical axis tended to be smaller in the LLIF at the final follow-up. Overall, perioperative and late complications were comparable in both procedures. However, LLIF procedures demonstrated significantly less intraoperative blood loss and a smaller incidence of postoperative epidural hematoma compared with PLIF/TLIF procedures in patients with ASD.

A recently identified Trichophyton rubrum major facilitator superfamily (MFS)-type transporter (TruMFS1) has been shown to give resistance to azole compounds and cycloheximide (CYH) when overexpressed in Saccharomyces cerevisiae. We investigated the roles of MFS1 in the intrinsic resistance of dermatophytes to CYH and chloramphenicol (CHL), which are commonly used to isolate these fungi, and to what extent MFS1 affects the susceptibility to azole antifungals. Susceptibility to antibiotics and azoles was tested in S. cerevisiae overexpressing MFS1 and ΔMFS1 mutants of Trichophyton benhamiae, a dermatophyte that is closely related to T. rubrum. We found that TruMFS1 functions as an efflux pump for CHL in addition to CYH and azoles in S. cerevisiae. In contrast, the growth of T. benhamiae ΔMFS1 mutants was not reduced in the presence of CYH but was severely impaired in the presence of CHL and thiamphenicol, a CHL analog. The suppression of MFS1 in T. benhamiae also increased the sensitivity of the fungus to fluconazole and miconazole. Our experiments revealed a key role of MFS1 in the resistance of dermatophytes to CHL and their high minimum inhibitory concentration for fluconazole. Suppression of MFS1 did not affect the sensitivity to CYH, suggesting that another mechanism was involved in resistance to CYH in dermatophytes.

Rapid defibrillation and high-quality cardiopulmonary resuscitation (CPR) are necessary for patients with cardiopulmonary arrest, one of the most serious and frequently encountered complications in cardiac catheterization laboratories. However, when the catheterization table is withdrawn from its neutral position for fluoroscopy, it is unstable and unsuitable for resuscitation because of its cantilever structure. To stabilize the table in its withdrawn position, the use of a table-stabilizing stick might improve CPR quality. To investigate the effect of using a cardiac catheterization table-stabilizing stick on CPR quality, a CPR simulation mannequin was placed on a cardiac catheterization table that was withdrawn from the C-arm of the X-ray machine. CPR quality was assessed with or without the use of a table-stabilizing stick under the table. The CPR quality assessment (Q-CPR) scores were 79.6 ± 11.4% using the table-stabilizing stick and 47.7 ± 30.3% without the use of the stick device (p = 0.02). In this simulation-based study, the use of a table-stabilizing stick in a cardiac catheterization table withdrawn from the C-arm of the X-ray machine improved the quality of CPR.

The ossification of the ligamentum flavum (OLF) was characterized by the replacement of the ligamentum flavum by ectopic new bone formation. Although OLF is well known as one of the causes of thoracic myelopathy through the compression of the spinal cord from the posterolateral side, contiguous multilevel OLF is quite a rare cause of myelopathy. Severe ossifications were reported that only present in individuals aged over 25 years. External triggers of OLF might likely be increased as a consequence of biomechanical stress to the effect of posttraumatic ossification. The authors described one rare case of an enormous contiguous multilevel OLF in a 20-year-old female’s thoracic spine after mild back trauma, to the extent that it was misdiagnosed as an old epidural hematoma initially. This female with obesity presented with a 6-month history of progressively worsening dorsal cord disorders. Resection or floating of the enormous OLF could be successfully achieved using O-arm-based navigation, and sensory loss, numbness, and gait disturbance were improved after operation.

The Suzuki–Miyaura coupling reaction is one of the most widely utilized C–C bond forming methods to create (hetero)biaryl scaffolds. The continuous-flow reaction using heterogeneous catalyst-packed cartridges is a practical and efficient synthetic method to replace batch-type reactions. A continuous-flow ligand-free Suzuki–Miyaura coupling reaction of (hetero)aryl iodides, bromides, and chlorides with (hetero)aryl boronic acids was developed using cartridges packed with spherical resin (tertiary amine-based chelate resin: WA30)-supported palladium catalysts (7% Pd/WA30). The void space in the cartridge caused by the spherical catalyst structures enables the smooth flow of a homogeneously dissolved reaction solution that consists of a mixture of organic and aqueous solvents and is delivered by the use of a single syringe pump. Clogging or serious backpressure was not observed.

In diffuse idiopathic hyperostosis (DISH), the ankylosed spine becomes susceptible to spinal fractures and spinal cord injuries due to the long lever arms of the fractured segments that make the fracture extremely unstable. The aim of this retrospective multicenter study was to examine the differences in DISH-affected spine fractures according to fracture level. The data of 285 cases with fractures of DISH-ankylosed segments diagnosed through computed tomography (CT) imaging were studied and the characteristics of 84 cases with cervical fractures were compared to 201 cases with thoracolumbar fractures. Examination of the CT images revealed that cervical fracture cases were associated with ossification of the posterior longitudinal ligament and had fractures at the intervertebral disc level, while thoracolumbar fracture cases were associated with ankylosing of the posterior elements and had fractures at the vertebral body. Neurologically, cervical fracture cases had a higher ratio of spinal cord injury leading to higher mortality, while thoracolumbar fracture cases had lower rates of initial spinal cord injury. However, a subset of thoracolumbar fracture cases suffered from a delay in diagnosis that led to higher rates of delayed neurological deterioration. Some of these thoracolumbar fracture cases had no apparent injury episode but experienced severe neurological deterioration. The information provided by this study will hopefully aid in the education of patients with DISH and raise the awareness of clinicians to potential pitfalls in the assessment of DISH trauma patients.

Angiolipomas are relatively rare benign tumors. Spinal angiolipomas that generally induce slow progressive cord compression are most commonly found in the thoracic region. A 49-year-old female with obesity presented with a 1-week history of progressively worsening back pain, paresthesia of lower limbs, and gait disturbance. When thoracic magnetic resonance imaging (MRI) revealed a dorsal epidural mass at the Th5–Th8 level, the patient underwent a laminectomy for gross total excision of the lesion. Both mature fatty tissue and abnormal proliferating vascular elements with thin or expanded walls were observed in the resected tumor. Nonfiltrating spinal angiolipoma was diagnosed and confirmed by pathology. After the operation, sensory loss, numbness, and gait disturbance were improved following the disappearing severe back pain. Following examinations indicated the absence of recurrence within 1 year. The angiolipomas of the spine are rare causes of spinal cord compression that generally induce slow progressive cord compression, but sudden onset or rapid worsening of neurological deterioration is observed in hemorrhagic spinal angiolipoma.

We developed a microwave-mediated continuous hydrogen production method from 2-PrOH using platinum on a spherical carbon-bead catalyst. The catalyst cartridge consisted of helical glass part, and straight glass part (helix−straight cartridge) was newly developed for the effective microwave heating of 2-PrOH in the presence of 5% Pt/CB. The microwave resonance was properly adjusted within 2.4−2.5 GHz using the helix−straight cartridge with the glass resonance-adjuster tube. The reaction was conducted by the irradiation of only 10 W of single-frequency microwaves and the catalyst was used continuously for at least 13 h without any loss of catalyst activity.

The catalyst activities of various heterogeneous palladium catalysts supported by anatase-, rutile- and brookite-type titanium oxide for ligand-free Suzuki−Miyaura cross-coupling reactions of aryl chlorides were evaluated. Palladium acetate [Pd(OAc)2], supported on anatase-type titanium oxide (TiO2) via acetonitrile solution impregnation process without reduction [Pd/TiO2 (anatase-type)], demonstrated the highest catalyst activity in comparison to those of other titanium oxide (rutile- or brookite-type) supported Pd(OAc)2 without reduction and reduced Pd/TiO2 (anatase-type) [Pd(red)/TiO2 (anatase-type)]. Various aryl chloride and bromide derivatives were smoothly coupled with arylboronic acids including heteroarylboronic acids in the presence of 5−10 mol% Pd/TiO2 (anatase-type) without the addition of any ligands. Although the fresh Pd/TiO2 (anatase-type) catalyst was surprisingly comprised of ca. 1:2 mixture of palladium(II) and palladium(0) species according to X-ray photoelectron spectroscopy (XPS), in spite of no reduction process, significant further increment of palladium(0) species was observed during the Suzuki−Miyaura coupling reaction, and Pd/TiO2 (anatase-type) was converted into a catalyst, which contained palladium(0) species as the main component [ca. 1:5 mixture of palladium(II) and palladium(0) species]. Therefore, the reduction via the electron donation process to the palladium(II) species may have occurred during the reaction on anatase-type titanium oxide.

We previously demonstrated the importance of quality management procedures for the handling of human bone marrow stromal cells (hBMSCs) and provided evidence for the existence of osteogenic inhibitor molecules in BMSCs. One candidate inhibitor is the ephrin type-A receptor 5 (EphA5), which is expressed in hBMSCs and upregulated during long-term culture. In this study, forced expression of EphA5 diminished the expression of osteoblast phenotypic markers. Downregulation of endogenous EphA5 by dexamethasone treatment promoted osteoblast marker expression. EphA5 could be involved in the normal growth regulation of BMSCs and could be a potential marker for replicative senescence. Although Eph forward signaling stimulated by ephrin-B-Fc promoted the expression of ALP mRNA in BMSCs, exogenous addition of EphA5-Fc did not affect the ALP level. The mechanism underlying the silencing of EphA5 in early cultures remains unclear. EphA5 promoter was barely methylated in hBMSCs while histone deacetylation could partially suppress EphA5 expression in early-passage cultures. In repeatedly passaged cultures, the upregulation of EphA5 independent of methylation could competitively inhibit osteogenic signal transduction pathways such as EphB forward signaling. Elucidation of the potential inhibitory function of EphA5 in hBMSCs may provide an alternative approach for lineage differentiation in cell therapy strategies and regenerative medicine.

Ossification of the posterior longitudinal ligament (OPLL) can cause severe and irreversible paralysis in not only the cervical spine but also the thoracolumbar spine. To date, however, the prevalence and distribution of OPLL in the whole spine has not been precisely evaluated in patients with cervical OPLL. Therefore, we conducted a multi-center study to comprehensively evaluate the prevalence and distribution of OPLL using multi-detector computed tomography (CT) images in the whole spine and to analyze what factors predict the presence of ossified lesions in the thoracolumbar spine in patients who were diagnosed with cervical OPLL by plain X-ray. Three hundred and twenty-two patients with a diagnosis of cervical OPLL underwent CT imaging of the whole spine. The sum of the levels in which OPLL was present in the whole spine was defined as the OP-index and used to evaluate the extent of ossification. The distribution of OPLL in the whole spine was compared between male and female subjects. In addition, a multiple regression model was used to ascertain related factors that affected the OP-index. Among patients with cervical OPLL, women tended to have more ossified lesions in the thoracolumbar spine than did men. A multiple regression model revealed that the OP-index was significantly correlated with the cervical OP-index, sex (female), and body mass index. Furthermore, the prevalence of thoracolumbar OPLL in patients with a cervical OP-index ≥ 10 was 7.8 times greater than that in patients with a cervical OP-index ≤ 5. The results of this study reveal that the extent of OPLL in the whole spine is significantly associated with the extent of cervical OPLL, female sex, and obesity.

UNLABELLED:We evaluated whether dexamethasone augments the osteogenic capability of bone marrow-derived stromal cells (BMSCs) and muscle tissue-derived stromal cells (MuSCs), both of which are thought to contribute to ectopic bone formation induced by bone morphogenetic protein-2 (BMP-2), and determined the underlying mechanisms. Rat BMSCs and MuSCs were cultured in growth media with or without 10-7 M dexamethasone and then differentiated under osteogenic conditions with dexamethasone and BMP-2. The effects of dexamethasone on cell proliferation and osteogenic differentiation, and also on ectopic bone formation induced by BMP-2, were analyzed. Dexamethasone affected not only the proliferation rate but also the subpopulation composition of BMSCs and MuSCs, and subsequently augmented their osteogenic capacity during osteogenic differentiation. During osteogenic induction by BMP-2, dexamethasone also markedly affected cell proliferation in both BMSCs and MuSCs. In an in vivo ectopic bone formation model, bone formation in muscle-implanted scaffolds containing dexamethasone and BMP-2 was more than two fold higher than that in scaffolds containing BMP-2 alone. Our results suggest that dexamethasone potently enhances the osteogenic capability of BMP-2 and may thus decrease the quantity of BMP-2 required for clinical application, thereby reducing the complications caused by excessive doses of BMP-2. HIGHLIGHTS:1. Dexamethasone induced selective proliferation of bone marrow- and muscle-derived cells with higher differentiation potential. 2. Dexamethasone enhanced the osteogenic capability of bone marrow- and muscle-derived cells by altering the subpopulation composition. 3. Dexamethasone augmented ectopic bone formation induced by bone morphogenetic protein-2.