Molecular Imaging of Labile Heme in Living Cells Using a Small Molecule Fluorescent Probe.

Labile heme (LH) is a complex of Fe(II) and protoporphyrin IX, an essential signaling molecule in various biological systems. Most of the subcellular dynamics of LH remain unclear because of the lack of efficient chemical tools for detecting LH in cells. Here, we report an activity-based fluorescence probe that can monitor the fluctuations of LH in biological events. H-FluNox is a selective fluorescent probe that senses LH using biomimetic N-oxide deoxygenation to trigger fluorescence. The selectivity of H-FluNox to LH is >100-fold against Fe(II), enabling the discrimination of LH from the labile Fe(II) pool in living cells. The probe can detect the acute release of LH upon NO stimulation and the accumulation of LH by inhibiting the heme exporter. In addition, imaging studies using the probe revealed a partial heme-export activity of the ATP-binding cassette subfamily G member 2 (ABCG2), potential LH pooling ability of G-quadruplex, and involvement of LH in ferroptosis. The successful use of H-FluNox in identifying fluctuations of LH in living cells offers opportunities for studying the physiology and pathophysiology of LH in living systems.

Lambertellin system, the mechanism for fungal replacement of Monilinia fructigena with Lambertella corni-maris without competitive inhibition on agar media.

The 'Lambertellin system' was disclosed which rationally explains the fungal replacement (mycoparasitism) of Monilinia fructigena (M. f., the host) with Lambertella corni-maris (L. corni-maris, the parasite) without competitive inhibition in the simultaneous incubations on agar media. The 'Lambertellin system' involves; (a) L. corni-maris secretes nontoxic lambertellols (1, 2) as the diffusible precursors of the authentic responsible substance 3 regardless of existence of the host M. f.; (b) In the absence of the host, the environment around the parasite is kept under neutral condition, and both 1 and 2 are readily transformed into 3; (c) Lambertellin (3) inhibits not only the host but also the parasite. The parasite degrades 3 for detoxification; and (d) Upon the host M. f. approaching closely to the area where the parasite inhabits, the environment around the parasite becomes acidic to stabilize 1 and 2, which gives them a chance to diffuse into the host area. Then these are gradually transformed into 3 to inhibit the host without damaging the parasite. This mechanism also accords with the progress of 'Natsu-Nenju' disease on apple fruits, which is known to be a mysterious phyto-disease because of two unique stages of its lifecycle, anamorphic (asexual) and teleomorphic (sexual). The 'Lambertellin system' would be categorized as a novel class of alleropathies.

Effects of α7 nicotinic acetylcholine receptor agonist against α-synuclein-induced neurotoxicity.

The α7 neuronal nicotinic acetylcholine receptor (α7 nAChR) is a potential target for the development of Parkinson's disease (PD) therapeutics. α-Synuclein (α-Syn), a principal component of Lewy bodies (cytoplasmic inclusions), is a major contributor to PD pathophysiology. Previous studies have demonstrated that activating α7 nAChR protects against nigrostriatal dopamine degeneration in acute and chronic PD animal models induced by 6-hydroxydopamine and rotenone, respectively. In the present study, we investigated the effects of PNU282987, a selective α7 nAChR agonist, against α-Syn-induced neurotoxicity in α-SynWT-, α-SynA30P-, and α-SynE46K-N2a cells. PNU282987 exhibited substantial neuroprotection against both wild-type and mutant-type α-Syn-induced toxicity. Furthermore, PNU282987 promoted transcription factor EB activity and reduced intracellular α-Syn protein levels through autophagy induction. These results highlight the therapeutic potential of α7 nAChR activation in diseases characterized by α-Syn aggregation, such as PD.

Functional outcome of a patient after hip disarticulation due to an infection 10 years after limb salvage surgery for osteosarcoma: A case report.

Limb salvage is a common procedure after extensive osteosarcoma resection. However, the long-term outcomes after limb salvage surgery (LSS) remain unclear. In this article, the case of a 24-year-old man who underwent hip disarticulation (HD) after an uncontrollable infection is presented. He was previously diagnosed with right distal femoral osteosarcoma and underwent LSS 10 years before disarticulation. Four years after LSS, an uncontrollable infection developed around the endoprosthesis, which eventually prompted HD. The Musculoskeletal Tumor Society (MSTS) functional rating system and the Toronto Extremity Salvage Score were used to compare the subject's activity statuses after LSS and HD. MSTS functional scores were 53.3% after LSS and 60% after HD. Toronto Extremity Salvage Scores were 78.3% after LSS and 95.8% after HD. The subject's emotional acceptance was 3 for LSS and 5 for HD (0 = worst and 5 = best). Both the MSTS and Toronto Extremity Salvage Scores were greater after HD than after LSS. The subject's improved emotional acceptance of the affected limb after HD contributed to the improved functional assessment scores. Alleviation of pain and other disabilities associated with the infection may have contributed to the higher functional scores after the more recent HD surgery. Even if amputation is unavoidable because of complications, high psychological acceptance may prevent a decrease in patient mobility and quality of life after amputation.

Effects and limitations of home-based motor-control exercise for chronic low back pain: A single center prospective study.

STUDY DESIGN: Prospective single-center observational study. OBJECTIVE: To investigate the effects and limitations of self-motor-control exercise in patients with chronic low back pain. SUMMARY OF BACKGROUND DATA: Although exercise therapy and physical therapy have been shown to be effective in treating chronic low back pain, these therapies are often discontinued due to patients' non-compliance, and their effectiveness cannot be fully demonstrated. METHODS: Fifteen patients with low back pain, no apparent organic disease, who had been symptomatic for at least three months, and could continue motor-control exercise at home for at least six months were included in the study. Low back pain (visual analog scale [VAS]), locomotor 25, stand-up test, two-step test, trunk and total body muscle mass by the impedance method, and spinal sagittal alignment were examined before the intervention to establish a baseline, and at two and six months after the intervention. RESULT: Significant improvement was observed in the back pain VAS (p<0.01), stand-up test (p = 0.03), two-step test (p = 0.01), and locomotor 25 (p = 0.04) before and after the intervention. In contrast, there were no significant changes in muscle mass and sagittal alignment. The effect of long-term exercise was more pronounced in patients without spinal deformity. CONCLUSIONS: Self-exercise for patients with chronic low back pain was effective in improving pain and function, although it did not directly affect muscle mass or alignment. Moreover, strength training of the lumbar back muscles alone was not found to be effective in patients with spinal deformities.

The neuroprotective effects of FG-4592, a hypoxia-inducible factor-prolyl hydroxylase inhibitor, against oxidative stress induced by alpha-synuclein in N2a cells.

Parkinson's disease (PD) is a neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra. The pathological hallmark of PD is the appearance of intraneuronal cytoplasmic α-synuclein (α-Syn) aggregation, called Lewy bodies. α-Syn aggregation is deeply involved in the pathogenesis of PD. Oxidative stress is also associated with the progression of PD. In the present study, to investigate whether a hypoxia-inducible factor (HIF)-prolyl hydroxylase (PH) inhibitor, FG-4592 (also called roxadustat), has neuroprotective effects against α-Syn-induced neurotoxicity, we employed a novel α-Syn stably expressing cell line (named α-Syn-N2a cells) utilizing a piggyBac transposon system. In α-Syn-N2a cells, oxidative stress and cell death were induced by α-Syn, and FG-4592 showed significant protection against this neurotoxicity. However, FG-4592 did not affect α-Syn protein levels. FG-4592 triggered heme oxygenase-1 (HO-1) expression downstream of HIF-1α in a concentration-dependent manner. In addition, FG-4592 decreased the production of reactive oxygen species possibly via the activation of HO-1 and subsequently suppressed α-Syn-induced neurotoxicity. Moreover, FG-4592 regulated mitochondrial biogenesis and respiration via the induction of the peroxisome proliferator-activated receptor-γ coactivator-1α. As FG-4592 has various neuroprotective effects against α-Syn and is involved in drug repositioning, it may have novel therapeutic potential for PD.

Absolute stereochemistry of altersolanol A and alterporriols.

The absolute stereochemistry of altersolanol A (1) was established by observing a positive exciton couplet in the circular dichroism (CD) spectrum of the C3,C4-O-bis(2-naphthoyl) derivative 10 and by chemical correlations with known compound 8. Before the discussion, the relative stereochemistry of 1 was confirmed by X-ray crystallographic analysis. The shielding effect at C7'-OMe group by C1-O-benzoylation established the relative stereochemical relationship between the C8-C8' axial bonding and the C1-C4/C1'-C4' polyol moieties of alterporriols E (3), an atropisomer of the C8-C8' dimer of 1. As 3 could be obtained by dimerization of 1 in vitro, the absolute configuration of its central chirality elements (C1-C4) must be identical to those of 1. Spectral comparison between the experimental and theoretical CD spectra supported the above conclusion. Axial stereochemistry of novel C4-O-deoxy dimeric derivatives, alterporriols F (4) and G (5), were also revealed by comparison of their CD spectra to those of 2 and 3.

Ten-year longitudinal investigation of astigmatism: The Yamagata Study (Funagata).

Despite numerous investigations into ocular or corneal astigmatism, the dynamic nature of astigmatism remains poorly understood. To reveal potential associations between age and astigmatism, 264 Japanese participants who underwent systemic and ophthalmological examinations in Funagata Town (Yamagata Prefecture, Japan) were evaluated over a 10-year period. Astigmatism was evaluated with regard to the cylinder power, cylinder axis, and vector analyses. Whereas the refractive cylinders showed age-related increases in patients in their 40s to 60s, the corneal cylinders did not change over 10 years. Nevertheless, cylindrical axis of the cornea demonstrated a continuous shift toward against-the-rule (ATR) astigmatism. Vector analyses revealed that the astigmatic shift toward ATR progressed continually after patients reached their 40s, although the shift did not accelerate with age. These novel insights may pave the way for the development of potential strategies for vision correction, including refractive surgeries, and vision-quality maintenance in the elderly.

Bunodosine 391: an analgesic acylamino acid from the venom of the sea anemone Bunodosoma cangicum.

A new acylamino acid, bunodosine 391 (BDS 391), was isolated from the venom of the sea anemone Bunodosoma cangicum. The structure was elucidated by spectroscopic analyses (2D NMR, ESIMS/MS) and verified by its synthesis. Intraplantar injection of BDS 391 into the hind paw of a rat induced a potent analgesic effect. This effect was not altered by naloxone (an opioid receptor antagonist), but was completely reversed by methysergide (a serotonin receptor antagonist), indicating that the effect is mediated by activation of serotonin receptors.

Effectiveness of Baricitinib in Refractory Seronegative Rheumatoid Arthritis and Uveitis: A Case Report.

Baricitinib is a Janus kinase (JAK) inhibitor used to treat refractory rheumatoid arthritis and blocks the subtypes JAK1 and JAK2. A 35-year-old man with seronegative rheumatoid arthritis complicated by bilateral severe non-granulomatous panuveitis was resistant to steroid treatment, multiple conventional disease-modifying antirheumatic drugs (methotrexate and salazosulfapyridine), and TNF-α inhibitors (adalimumab and infliximab). Therefore, the TNF-α inhibitors were switched to baricitinib to decrease the activity of systemic arthritis. Along with the amelioration of inflammatory activity in seronegative rheumatoid arthritis, the inflammatory activity of uveitis was decreased. Vitreous opacity, serous retinal detachment, and anterior chamber cells showed improvement. Baricitinib was effective not only in refractory systemic arthritis but also in uveitis, which may provide a new treatment option for patients with refractory uveitis.

A Novel Exercise Facilitation Method in Combination with Cognitive Behavioral Therapy Using the Ikiiki Rehabilitation Notebook for Intractable Chronic Pain: Technical Report and 22 Cases.

Recent clinical practice guidelines for chronic pain indicate, with a high evidence level, that the combination of exercise and cognitive behavioral therapy (CBT) is effective. The purpose of this study was to evaluate the effectiveness of an exercise facilitation method in combination with CBT using the "Ikiiki Rehabilitation Notebook" for patients with intractable chronic pain. "Ikiiki" means active in Japanese. A total of 22 cases with chronic low back (n = 13), lower extremity (n = 8), or neck (n = 1) pain were treated using this notebook. Two cases dropped out, leaving 22 cases. Each case was evaluated in terms of the numerical rating scale (NRS) of the pain, activities of daily living (ADL), pain catastrophizing scale (PCS), and quality of life (QOL) at pretreatment and post-treatment. The endpoint of the method was to achieve the long-term goals set by the patients. The mean treatment period was 11.2 months. The outcomes were as follows: improvement of presenteeism: nine cases; enhanced participation in hobbies: seven cases; improved school attendance: two cases; return to work: one case; improvement of self-care and/or self-efficacy: three cases. The NRS, ADL, PCS, and QOL were significantly improved after the treatment. This method is possibly valuable for educating patients about the cause and treatment of chronic pain and actively facilitating exercise and social participation. Further studies are needed to investigate the effectiveness of using this notebook for the patient with intractable chronic pain.

Age-Related Changes in Astigmatism and Potential Causes.

Astigmatism causes deterioration of the retinal image and affects vision quality. Maintenance and improvement of visual function requires an understanding of the prevalence, age-related changes, and mechanisms of astigmatism. In this article, we discuss the findings of studies that investigated astigmatism. Some of these studies showed that the prevalence of high degrees of astigmatism in childhood typically decreases with emmetropization. With-the-rule astigmatism occurs most commonly in young adults. With age, the prevalence of astigmatism increases, and the axis shifts from a predominance of with-the-rule astigmatism to a predominance of against-the-rule astigmatism. This age-related change is caused by alterations in corneal curvature. Although the cause of this change is not fully understood, alterations in the position and tension of the eyelid, corneal stromal collagen fibrils, Descemet membrane, and extraocular muscles may influence the shape of the cornea. Furthermore, genetic factors may contribute to the development of astigmatism. Technological advances in ophthalmology are expected to improve our understanding of the etiology of astigmatism and enable the maintenance of quality of vision.

Biosynthetic studies of spiroleptosphol.

Studies of the biosynthesis of spiroleptosphol (1) revealed that 1 comprised a heptaketide (C1, C5-C10, and C12-C18 moiety), two methyl carbons (C19 and C20) from methionine, and a C(3) unit (C3, C4, and C11 moiety) derived through the TCA cycle.

Four analogues of spiroleptosphol isolated from Leptosphaeria doliolum.

Spiroleptosphol B (2), spiroleptosphol C (3), norleptosphol C (4) and hydroleptosphol (5) were isolated from ascomycete Leptosphaeria doliolum. Detailed (1)H and (13)C NMR spectral analyses revealed these were structural analogues of spiroleptosphol (1) which we have recently isolated from the same fungi. Spiroleptosphol B (2) carried an unprecedent 5,3-dioxatricyclo[4.4.0.1(1.4)]undecane framework in place of the spirobicyclo ring system of 1. Spiroleptosphol C (3) was a 17-(R)-hydroxy derivative of 1. Norleptosphol C (4) was deduced to be the monocyclic structure biosynthetically resulted by decarboxylation from 3. Although 5 gave broaden (1)H NMR spectrum, it was gradually transformed to 2 which suggested being a hydrolysate of 1.

Isolation and absolute stereochemistry of optically active sydonic acid from Glonium sp. (Hysteriales, Ascomycota).

Optically active sydonic acid (1) was isolated for the first time from a culture broth of Glonium sp. The absolute stereochemistry was established to be (S) by comparing the circular dichroism (CD) spectrum with that of (+)-curcutetraol after conversion into (+)-sydonol (2).

Evidence-Based Recommendations on the Pharmacological Management of Osteoarthritis and Chronic Low Back Pain: An Asian Consensus

The overall burden of chronic musculoskeletal pain in Asian countries will continue to increase as the population ages, as will the demand for safe and effective pain management. Currently available Asian guidelines are mostly outdated and targeted only to primary care. Implementation of international guidelines may be unsuitable for Asian patients due to cultural, local economic and regulatory factors. With the aim of developing Asian-specifi c consensus recommendations for the pharmacological management of osteoarthritis (OA) pain and chronic low back pain (cLBP), we convened to review and discuss recent available evidence for pharmacotherapy, clinical experiences, and current practice challenges they face in the region, including challenges in opioid use. Taking these into consideration, we provided general recommendations for the overall assessment and management of OA pain and cLBP. The strength of the recommendations regarding the use of pharmacological agents was assessed using the Grades of Recommendation Assessment, Development and Evaluation (GRADE) system. Where evidence is confl icting or limited, we made no recommendation pending the availability of further evidence. We recommend topical non-steroidal anti-infl ammatory drugs (NSAIDs) as a fi rst-line pharmacological treatment of OA pain, while oral NSAIDs should be considered as a fi rst-line pharmacological treatment of cLBP. Acetaminophen has been commonly used as the fi rst-line treatment for OA pain and cLBP, but its long-term use is not recommended based on recent evidence. These consensus recommendations are not prescriptive, and serve as a guide for decision-making in clinical practice. The optimal management of OA pain and cLBP should ultimately be individualized to each patient.

Syntheses of lambertellols and their stable analogues; investigation of the real active species in the mycoparasitism by Lambertella species.

Lambertellols A (1) and B (2), isolated from mycoparasites Lambertella species, were synthesized. The synthesis features intramolecular aldol-type cyclizations of aldehydes 12 and 14 and site specific oxidations of 1-hydroxylambertellols as key steps. The synthesis also provided all diastereomers of 1-hydrolambertellols 17-19 and 25. Chiral resolution made the optically active forms available, which enabled the investigation of the real active species in the mycoparasitism by Lambertella species against Monilinia fructigena. These experiments suggested that lambertellin (3) is responsible for this phenomenon. Chemically labile 1 and 2 should be converted to 3 during the bioassay. The parasite may excrete 1 and 2 as readily diffusible forms, which are then transformed into 3 to inhibit the host M. fructigena. The parasite may have acquired this "drug delivery system" mechanism as an evolutionary enhancement.

Structure and biosynthesis of norneolambertellin produced by Lambertella sp. 1346.

Norneolambertellin (1) was isolated from a mycoparasite Lambertella sp. 1346. Combined analysis of the NMR spectra and chemical shift prediction based on molecular orbital calculation successfully revealed a novel pyrano[3,2-c]chromene-2,5-dione structure, which was further confirmed by X-ray crystallographic analysis. Isotopomer distribution analysis of the sample, prepared under labeling conditions, deduced its biosynthetic pathway.

Spiroleptosphol isolated from Leptosphaeria doliolum.

A novel gamma-methylidene-spirobutanolide spirolephtoshol (1) was isolated from ascomycetous fungus Leptosphaeria doliolum as a cytotoxic compound. The relative structure was established by the NMR analysis involving the NOE experiments. Absolute structure of the bicyclic moiety was determined by chemical derivation followed by the CD analysis. The relative and absolute stereochemistry of the side chain was established by comparison of the (1)H NMR spectra and the chiral GC chromatograms of the degradation product with the synthetic samples.

Age-related variations in corneal geometry and their association with astigmatism: The Yamagata Study (Funagata).

To investigate how aging affects corneal geometry in Japanese adults, and the association between corneal geometry and astigmatism.We included 421 participants who had undergone systemic and ophthalmological examinations in 2015 in Funagata town, Yamagata, Japan. Corneal topographic data were obtained using anterior-segment optical coherence tomography (CASIA SS-1000). Astigmatism was evaluated using power vector analyses where J0 represents the power of the orthogonal astigmatism. Positive values of J0 indicate with-the-rule astigmatism, while negative values indicate against-the-rule (ATR) astigmatism.Regarding age-related variations in corneal geometry, the anterior elevations at axis 0° and 180° decreased, and those at axis 90° and 270° increased with increasing age in linear regression analyses, demonstrating horizontal steepening and vertical flattening of the corneal surface. There were no significant age-related variations in posterior elevations and pachymetry findings, including central corneal thickness. Regarding age-related variations in orthogonal astigmatism, the mean values of J0 and corneal J0 (cJ0) decreased by -0.014 and -0.015 per year of increase in age, indicating astigmatic shift toward ATR. Regarding the correlation between corneal geometry and astigmatism, the shift toward ATR was positively correlated with horizontal steepening and vertical flattening, in accordance with the age-related corneal variations. In addition, the posterior surface of the cornea also has an association with this shift to some extent.The results of our population-based study demonstrated that the age-related variation in astigmatism is associated with geometrical changes in the cornea, especially those in the anterior surface of the cornea.