Comparison of Postoperative Complications and Outcomes in Anterior Cervical Spine Surgery: Ossification of the Posterior Longitudinal Ligament Versus Cervical Spondylotic Myelopathy.

STUDY DESIGN: Retrospective cohort study. OBJECTIVES: To compare the frequency of complications and outcomes between patients with ossification of the posterior longitudinal ligament (OPLL) of the cervical spine and those with cervical spondylotic myelopathy (CSM) who underwent anterior surgery. SUMMARY OF BACKGROUND DATA: Anterior cervical spine surgery for OPLL is an effective surgical procedure; however, it is complex and technically demanding compared with the procedure for CSM. Few reports have compared postoperative complications and clinical outcomes after anterior surgeries between the 2 pathologies. METHODS: Among 1434 patients who underwent anterior cervical spine surgery at 3 spine centers within the same spine research group from January 2011 to March 2021, 333 patients with OPLL and 488 patients with CSM were retrospectively evaluated. Demographics, postoperative complications, and outcomes were reviewed by analyzing medical records. In-hospital and postdischarge postoperative complications were investigated. Postoperative outcomes were evaluated 1 year after the surgery using the Japanese Orthopaedic Association score. RESULTS: Patients with OPLL had more comorbid diabetes mellitus preoperatively than patients with CSM ( P <0.001). Anterior cervical corpectomies were more often performed in patients with OPLL than in those with CSM (73.3% and 14.5%). In-hospital complications, such as reoperation, cerebrospinal fluid leak, C5 palsy, graft complications, hoarseness, and upper airway complications, occurred significantly more often in patients with OPLL. Complications after discharge, such as complications of the graft bone/cage and hoarseness, were significantly more common in patients with OPLL. The recovery rate of the Japanese Orthopaedic Association score 1 year postoperatively was similar between patients with OPLL and those with CSM. CONCLUSION: The present study demonstrated that complications, both in-hospital and after discharge following anterior spine surgery, occurred more frequently in patients with OPLL than in those with CSM.

The Ptk2-Pma1 pathway enhances tolerance to terbinafine in Trichophyton rubrum.

The increasing prevalence of dermatophyte resistance to terbinafine, a key drug in the treatment of dermatophytosis, represents a significant obstacle to treatment. Trichophyton rubrum is the most commonly isolated fungus in dermatophytosis. In T. rubrum, we identified TERG_07844, a gene encoding a previously uncharacterized putative protein kinase, as an ortholog of budding yeast Saccharomyces cerevisiae polyamine transport kinase 2 (Ptk2), and found that T. rubrum Ptk2 (TrPtk2) is involved in terbinafine tolerance. In both T. rubrum and S. cerevisiae, Ptk2 knockout strains were more sensitive to terbinafine compared with the wild types, suggesting that promotion of terbinafine tolerance is a conserved function of fungal Ptk2. Pma1 is activated through phosphorylation by Ptk2 in S. cerevisiae. Overexpression of T. rubrum Pma1 (TrPma1) in T. rubrum Ptk2 knockout strain (ΔTrPtk2) suppressed terbinafine sensitivity, suggesting that the induction of terbinafine tolerance by TrPtk2 is mediated by TrPma1. Furthermore, omeprazole, an inhibitor of plasma membrane proton pump Pma1, increased the terbinafine sensitivity of clinically isolated terbinafine-resistant strains. These findings suggest that, in dermatophytes, the TrPtk2-TrPma1 pathway plays a key role in promoting intrinsic terbinafine tolerance and may serve as a potential target for combinational antifungal therapy against terbinafine-resistant dermatophytes.

First atroposelective Chan-Lam coupling for the synthesis of C-N linked biaryls.

The first atroposelective Chan-Lam coupling for the synthesis of C-N axial enantiomers is reported with good yields and ee. MnO2 additive is crucial for the success of the coupling. The longstanding problem of the lack of enantioselective synthesis to make chiral C-N linked atropisomers is solved.

Is milbemycin a game changer against antifungal drug-resistant mycoses?

Milbemycin oximes are macrocyclic lactones that have a broad spectrum of activity against nematode infection in animals. They are known to block drug efflux, which increases the susceptibility of fungi to azoles. We investigated the effects of milbemycin on the azole susceptibility of fungi (Aspergillus fumigatus, Candida albicans, C. auris, Cryptococcus neoformans, and Trichophyton rubrum). To screen for changes in azole susceptibility, fungal growth was tested on a culture medium containing 1 µg/ml milbemycin. The results showed that milbemycin increased the azole susceptibility of azole-resistant strains of C. albicans, C. auris, C. neoformans, and T. rubrum. Thus, milbemycin might be useful against antifungal drug-resistant strains.

Milbemycin blocks drug efflux and increases the azole susceptibility of azole-resistant strains of Candida albicans, C. auris, Cryptococcus neoformans, and Trichophyton rubrum. This drug is expected to be a game changer against antifungal drug-resistant infections.

[Questionnaire Survey on IVR Dose Management in Tokai Region].

PURPOSE: National diagnostic reference levels in Japan 2020 (DRLs 2020) have been published. In the field of angiography, in addition to the fluoroscopic dose rate, incident air kerma at the patient entrance reference point displayed on the equipment (Ka,r: mGy) and air kerma-area product displayed on the equipment (PKA: Gycm2) were set. A questionnaire survey was conducted at each facility in the Tokai region to confirm the status of medical radiation dose control in the region. METHOD: A questionnaire survey was conducted at each facility in the Tokai region. The items were fluoroscopic dose rate in each area (head and neck, cardiac, chest and abdomen, and limbs), DA and DSA dose rates, and dose area product meter (Ka,r, PKA) for the main procedures in each area. RESULT: The median values in this study were lower than those in the DRLs 2020, indicating that appropriate dose control is being implemented in the Tokai region. The trends of fluoroscopic and radiographic dose rates were different in each area, and there was some variation among the facilities. CONCLUSION: We believe that the incorporation of fluoroscopic and radiographic dose rates by area into the DRLs will facilitate more appropriate dose control at each facility in the future.

Oxidative Functionalization of Catechol Derivatives Substituted with Electron-Withdrawing Groups.

Catechols possessing electron-withdrawing groups at the C3 position effectively underwent oxidative functionalization at the C4 position in the presence of phenyliodine(III) diacetate (PIDA) and heteroarene nucleophiles (e.g., indole, indazole, and benzotriazole) to produce the corresponding biaryl products. The PIDA-mediated oxidation of catechol derivatives afforded the ortho-benzoquinone intermediate, which subsequently underwent regioselective nucleophilic addition to the α,ß-unsaturated carbonyl moiety of ortho-benzoquinone using indole, indazole, and benzotriazole to give 4-substituted catechol derivatives in a one-pot manner. Notably, the nucleophilic substitution positions of indazole and benzotriazole were perfectly controlled. Additionally, the reaction using N-methylaniline as the nucleophile afforded a tertiary amine product.

TrCla4 promotes actin polymerization at the hyphal tip and mycelial growth in Trichophyton rubrum.

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.

Two different types of tandem sequences mediate the overexpression of TinCYP51B in azole-resistant Trichophyton indotineae.

Trichophyton indotineae is an emerging dermatophyte that causes severe tinea corporis and tinea cruris. Numerous cases of terbinafine- and azole-recalcitrant T. indotineae-related dermatophytosis have been observed in India over the past decade, and cases are now being recorded worldwide. Whole genome sequencing of three azole-resistant strains revealed a variable number of repeats of a 2,404 base pair (bp) sequence encoding TinCYP51B in tandem specifically at the CYP51B locus position. However, many other resistant strains (itraconazole MIC ≥0.25 µg/mL; voriconazole MIC ≥0.25 µg/mL) did not contain such duplications. Whole-genome sequencing of three of these strains revealed a variable number of 7,374 bp tandem repeat blocks harboring TinCYP51B. Consequently, two types of T. indotineae azole-resistant strains were found to host TinCYP51B in tandem sequences (type I with 2,404 bp TinCYP51B blocks and type II with 7,374 bp TinCYP51B blocks). Using the CRISPR/Cas9 genome-editing tool, the copy number of TinCYP51B within the genome of types I and II strains was brought back to a single copy. The azole susceptibility of these modified strains was similar to that of strains without TinCYP51B duplication, showing that azole resistance in T. indotineae strains is mediated by one of two types of TinCYP51B amplification. Type II strains were prevalent among 32 resistant strains analyzed using a rapid and reliable PCR test.

Long Amplification PCR (LA-PCR) Detection of Azole Resistant Trichophyton indotineae.

Trichphyton indotineae, a species newly designated in 2020 independent of T. interdigitale, comprises highly terbinafine (TRF)-resistant dermatophytosis that is epidemic in North India and spreding to worldwide. Some clinical isolates of T. indotineae have been resistance both TRF and azoles that might be caused the treatment failure. To detect the azole resistance strains, we developed a long amplification PCR (LA-PCR) detection method for the tandem repeat of the CYP51B (encoding sterol 14a-demethylase gene) in T. indotineae. Contrasting the drug susceptibility test results with the LA-PCR results confirmed a trend toward low susceptibility to azole antifungal agents in strains with amplifications of 9.5 kbp or greater (3 or more copies of CYP51B). Our results suggest that the method could be detected rapidly of low-susceptibility strains to azole antifungal agents.

Microwave-assisted C-C bond formation of diarylacetylenes and aromatic hydrocarbons on carbon beads under continuous-flow conditions.

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.

Impact of COVID-19 on the Surrounding Environment of Nursing Home Residents and Attitudes towards Infection Control and Oral Health Care among Nursing Home Staff in Japan.

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.

Pelvic incidence is a risk factor for lower instrumented vertebra failure in adult spinal deformity patients who underwent corrective fusion terminating at the L5 vertebra.

BACKGROUND: Factors related to postoperative mechanical failure after long fusion with lower instrumented vertebra (LIV) at L5 have not been well investigated. Elucidating such factors may allow us to perform alternatives to spinopelvic fusion for adult spinal deformity (ASD) cases. We investigated the incidence and risk factors of LIV failure in patients with ASD who underwent surgical treatment of long corrective fusion until the L5 vertebrae. METHODS: Between 2009 and 2018, 52 patients who underwent corrective fusions to L5 were followed-up for at least one-year. We evaluated the associated patient factors for LIV failure which include loosening of the pedicle screw of LIV, fracture of LIV, distal junctional kyphosis (DJK). RESULTS: The mean age of the participants was 71.2 ± 7.59 (range, 44-84). LIV failure occurred in 20 patients (38.5%), and 6 patients (11.5%) underwent secondary surgery for caudal segments. The mean pelvic incidence (PI) was 52.5 ± 9.8 in the failure group versus 45.3 ± 11.4 in non-failure group (P = 0.02) and pelvic tilt (PT) was 39.1 ± 9.0 versus 32.4 ± 13.0. There were no significant differences in sex, age, body mass index, number of levels fused, and other radiographic data. Logistic regression analysis that included T1 pelvic angle, PT, PI - postoperative LL and PI also identified PI as the only significant determinant of LIV failure (OR = 1.07, P = 0.034). Receiver operating characteristic analysis demonstrated that a PI over 50.0° was associated with LIV failure (sensitivity 63%, specificity 70%, AUC 0.694). CONCLUSION: LIV failure was frequently observed after long corrective fusion for patients with ASD. High PI was found to be a significant risk factor for the LIV failure.

K-Line Tilt is a Predictor of Postoperative Kyphotic Deformity After Laminoplasty for Cervical Myelopathy Caused by Ossification of the Posterior Longitudinal Ligament.

STUDY DESIGN: Retrospective single-center study. OBJECTIVES: K-line is a decision-making tool to determine the appropriate surgical procedures for patients with cervical ossification of the posterior longitudinal ligament (C-OPLL). Laminoplasty (LAMP) is one of the standard surgical procedures indicated on the basis of K-line measurements (+: OPLL does not cross the K-line). We investigated the impact of K-line tilt, a radiographic parameter of cervical sagittal balance measured using the K-line, on surgical outcomes after LAMP. METHODS: The study included 62 consecutive patients with K-line (+) C-OPLL who underwent LAMP. The following preoperative and postoperative radiographic measurements were evaluated: (1) the K-line, (2) K-line tilt (an angle between the K-line and vertical line), (3) center of gravity of the head -C7 sagittal vertical axis, (4) C2-C7 lordotic angle, (5) C7 slope, and (6) C2-C7 range of motion. Clinical results were evaluated using the Japanese Orthopedic Association scoring system for cervical myelopathy (C-JOA score). RESULTS: All the patients had non-kyphotic cervical alignment (CL ≥ 0°) preoperatively; however, kyphotic deformity (CL < 0°) was observed in 6 patients (9.7%) postoperatively. The recovery rate of the C-JOA scores was poor in the kyphotic deformity (+) group (7.8%) than in the kyphotic deformity (-) group (47.5%). The K-line tilt was identified to be a preoperative risk factor in the multivariate analysis, and the cutoff K-line tilt for predicting the postoperative kyphotic deformity was 20°. CONCLUSIONS: LAMP is not suitable for K-line (+) C-OPLL patients with K-line tilts >20°.

Predictors of good outcome after anterior decompression and fusion in patients with ossification of the posterior longitudinal ligament of the thoracic spine.

BACKGROUND: There are few reports concerning determinants of the surgical outcome of anterior decompression and fusion (ADF) when performed for ossification of the posterior longitudinal ligament of the thoracic spine (T-OPLL). We investigated prognostic factors associated with neurological recovery in the patients with T-OPLL. METHODS: This retrospective study included consecutive cases of T-OPLL patients from January 2002 to January 2020 and minimum one-year follow-up. Data were collected for sex, age, body mass index, preoperative manual muscle test score for the weakest muscle, surgical data, and preoperative and postoperative findings on radiographs, magnetic resonance images, and computed tomography scans. Imaging data were also collected, including preoperative kyphotic angles, canal occupancy ratio, type of OPLL, and high-intensity areas on T2-weighted images. The Japanese Orthopaedic Association score for thoracic myelopathy (T-JOA) was used to evaluate the recovery of the thoracic myelopathy. The patients were categorized according to whether the improvement in T-JOA score was >50% or ≤50%. RESULTS: Forty-six patients who underwent anterior procedures for T-OPLL were included in the study. Preoperative and postoperative T-JOA scores were 4.2 ± 2.3 and 7.9 ± 2.1, respectively. The improvement in the T-JOA score was 54.5 ± 25.6%. The proportion of patients with beak-type OPLL was significantly higher in the >50% JOA improvement group (23/27) than in the ≤50% group (9/19) (p = 0.009) and the canal occupancy ratio was significantly lower in the >50% group (56.3 ± 12.2% vs 64.4 ± 8.73%; p = 0.0163). There were no significant between-group differences in other factors. CONCLUSIONS: Beak-type ossification and a low canal occupancy ratio are predictors of good outcome after ADF in patients with T-OPLL. ADF should be considered in patients with either or both of these features.

Tacrolimus inhibits stress responses and hyphal formation via the calcineurin signaling pathway in Trichosporon asahii.

The pathogenic fungus Trichosporon asahii causes fatal deep-seated mycosis in immunocompromised patients. Calcineurin, which is widely conserved in eukaryotes, regulates cell growth and various stress responses in fungi. Tacrolimus (FK506), a calcineurin inhibitor, induces sensitivity to compounds that cause stress on the cell membrane and cell wall integrity. In this study, we demonstrated that FK506 affects stress responses and hyphal formation in T. asahii. In silico structural analysis revealed that amino acid residues in the binding site of the calcineurin-FKBP12 complex that interact with FK506 are conserved in T. asahii. The growth of T. asahii was delayed by FK506 in the presence of SDS or Congo red but not in the presence of calcium chloride. FK506 also inhibited hyphal formation in T. asahii. A mutant deficient of the cnb gene, which encodes the regulatory subunit B of calcineurin, exhibited stress sensitivities on exposure to SDS and Congo red and reduced the hyphal forming ability of T. asahii. In the cnb-deficient mutant, FK506 did not increase the stress sensitivity or reduce hyphal forming ability. These results suggest that FK506 affects stress responses and hyphal formation in T. asahii via the calcineurin signaling pathway.

Establishment of a gene recombination method for a major human skin commensal fungus, Malassezia restricta, using Agrobacterium tumefaciens-mediated gene transfer system.

Malassezia restricta is the most predominant fungus in the microbiome of human skin. This microorganism can cause or exacerbate Malassezia-associated skin dermatitis, seborrheic dermatitis, atopic dermatitis, and pityriasis versicolor. The virulence factors of M. restricta have not been analyzed because a gene recombination system has not been developed. In this study, we established an Agrobacterium tumefaciens-mediated gene transfer (ATMT) system, optimized for generating gene-deficient mutants of M. restricta. A mutant of FKB1 gene, which encodes the FKBP12 protein that binds to the calcineurin inhibitor tacrolimus, was generated using the ATMT system. Subsequently, the FKB1 gene was reintroduced into the FKB1 gene-deficient mutant for obtaining a gene-complemented strain. The wild-type strain of M. restricta was sensitive to tacrolimus, whereas the FKB1 gene-deficient mutant was resistant to tacrolimus; the phenotypic drug susceptibility in the mutant was restored by reintroducing the FKB1 gene. Contrastingly, the FKB1 gene-deficient mutant was not resistant to cyclosporine A, which also inhibits calcineurin by binding to cyclophilin A. The gene recombination system for M. restricta will facilitate in elucidating the molecular mechanisms causing Malassezia-associated dermatitis.

A critical role of calcineurin in stress responses, hyphal formation, and virulence of the pathogenic fungus Trichosporon asahii.

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.

Effect of Topical Antifungal Luliconazole on Hyphal Morphology of Trichophyton mentagrophytes Grown on in vitro Onychomycosis Model.

Luliconazole, recently launched in Japan, is a novel topical imidazole antifungal agent for the treatment of onychomycosis. Using in vitro onychomycosis model, the effect of luliconazole on the morphology of the growing hyphae of Trichophyton mentagrophytes was investigated by scanning electron microscopy (SEM). The model was produced by placing human nail pieces on an agar medium seeded with conidia of T. mentagrophytes. After incubating the agar medium for 3 days, luliconazole was applied to the surface of the nail in which hyphal growth was recognized, then cultured for up to 24 h. The initial change after treatment with the drug was the formation of fine wrinkles on the surface of the hyphae, eventually, the hyphae were flattened, and after that, no hyphal growth was observed. On the other hand, when the nails were pretreated with luliconazole for 1 h, no hyphal growth was observed even after culturing for 24 h. This study suggests that luliconazole has a strong antifungal activity by inhibiting the ability of fungi to grow and the drug has both excellent nail permeation and retention properties.

A joint PCR-based gene-targeting method using electroporation in the pathogenic fungus Trichosporon asahii.

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.