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The technique most widely used to quantitatively measure leg edema is only a pitting edema method. It has recently become possible to digitize B-mode ultrasound images and accurately quantify their brightness using an image-analysis software program. The purpose of this study was to find new indices of the grade of leg skin, to study whether or not analyses of the subcutaneous layer of leg skin on ultrasound images using image-editing software program can be used to evaluate it and to digitize it. Images of 282 subcutaneous layers of leg skin in 141 pregnant women were obtained using a B-scan portable ultrasound device. Rectangular photographs (vertical: skin thickness; horizontal: width of probe) were obtained using an image-editing program, and the luminous intensity (pixel grayscale: 0–255) and thickness of the skin were calculated using a histogram. We investigated the correlation between these parameters and the grade of pitting edema (0–3). There was a significant positive correlation between the grade of pitting edema and the average luminous intensity value, its standard deviation, and the skin thickness (ρ = 0.36, ρ = 0.22, ρ = 0.51, p < 0.0001, respectively). In particular, there was strong positive correlation between the grade of pitting edema and both the total number of pixels in a rectangle × (multiplied by) the average luminous intensity value and the total number of pixels in a rectangle × the standard deviation of the average luminous intensity value (ρ = 0.58 and ρ = 0.59, p < 0.0001, respectively). We could quantitatively evaluate the grade of leg edema by analyzing ultrasound photographs of the subcutaneous layer of the leg skin using an image-editing software program and found new indices to digitize it. [ABSTRACT FROM AUTHOR]

A rotary motor [F.sub.1], a catalytic part of ATP synthase, makes a 120[degrees] step rotation driven by hydrolysis of one ATP, which consists of 80[degrees] and 40[degrees] substeps initiated by ATP binding and probably by ADP and/or [P.sub.i] dissociation, respectively. During active rotations, [F.sub.1] spontaneously fails in ADP release and pauses after a 80[degrees] substep, which is called the ADP-inhibited form. In the present work, we found that, when pushed >+40[degrees] with magnetic tweezers, the pausing [F.sub.1] resumes its active rotation after releasing inhibitory ADP. The rate constant of the mechanical activation exponentially increased with the pushed angle, implying that [F.sub.1] weakens the affinity of its catalytic site for ADP as the angle goes forward. This finding explains not only its unidirectional nature of rotation, but also its physiological function in ATP synthesis; it would readily bind ADP from solution when rotated backward by an [F.sub.o] motor in the ATP synthase. Furthermore, the mechanical work for the forced rotation was efficiently converted into work for expelling ADP from the catalytic site, supporting the tight coupling between the rotation and catalytic event. ADP inhibition | ATP synthase | F1-ATPase | magnetic tweezers | single-molecule observation

Aim: The effects of detergent, ethanol and ethanol with plant meadowfoam oil on the growth of the red heterobasidomycete Xanthophyllomyces dendrorhous and on the production of astaxanthin (3,3′‐dihydroxy‐β,β‐carotene‐4,4′‐dione) and fatty acids in this red yeast were investigated. Methods and Results: Ethanol supplementation at a final concentration of 0.8% (v/v) caused an increase in the growth, astaxanthin production and fatty acid production of treated X. dendrorhous compared with untreated X. dendrorhous. Supplementation of meadowfoam oil with 0.8% ethanol further improved the growth and astaxanthin production of X. dendrorhous. Fatty acid compositions following supplementation with various concentrations of ethanol and oil were also analysed. With 0.8% ethanol supplementation, the ratio of linoleic acid (C18:2) and α‐linolenic acid (C18:3ω3, ALA) decreased. Conversely, with 1.8% ethanol supplementation, the ALA ratio increased. Conclusions: Ethanol can serve as a promoting factor for coproduction of astaxanthin and fatty acids in X. dendrorhous, whereas simultaneous supplementation of ethanol and meadowfoam oil can cause further astaxanthin production. Significance and Impact of Study: Astaxanthin is widely used in various functional products because of its antioxidant activity. This study shows that X. dendrorhous can coproduce astaxanthin and functional fatty acids at high levels following supplementation with ethanol. [ABSTRACT FROM AUTHOR]

[F.sub.1]-ATPase is a rotary motor enzyme in which a single ATP molecule drives a 120 [degrees] rotation of the central [gamma] subunit relative to the surrounding [[alpha].sub.3][[beta].sub.3] ring. Here, we show that the rotation of [F.sub.1]-ATPase spontaneously lapses into long ([approximately equal to] 30 s) pauses during steady-state catalysis. The effects of ADP-Mg and mutation on the pauses, as well as kinetic comparison with bulk-phase catalysis, strongly indicate that the paused enzyme corresponds to the inactive state of [F.sub.1]-ATPase previously known as the ADP-Mg inhibited form in which [F.sub.1]-ATPase fails to release ADP-Mg from catalytic sites. The pausing position of the [gamma] subunit deviates from the ATP-waiting position and is most likely the recently found intermediate 90 [degrees] position.

Abstract Background Bullous keratopathy (BK), a severe sight-threatening disorder can have a variety of etiologies such as prophylactic laser iridotomy, intraocular surgery, trauma, and other ocular disorders. However, there are cases of unknown origins, among which a unique clinical entity namely pseudoexfoliation syndrome (PEX) is having increased importance. Case presentation In this case note, we report the clinical features and in vivo confocal microscopic and pathological findings of two BK cases of unknown cause. Conclusions Our findings suggest that the BK was caused by the corneal endotheliopathy of PEX, a common disease that could affect up to 30% of people over 60 years old and is more prevalent than we have believed.

This paper describes a method for the rapid exchange of buffer solution during single-molecule observation. We use a simple, transparent, all-plastic one-way valve integrated on a coverslip on a flow cell. The valve is formed using a membrane made of parylene covering a microhole. It opens when a buffer solution is introduced from the microhole (the flow pushes the cover membrane) and closes when suctioning the buffer solution (the membrane is pulled back and seals the microhole to prevent the diffusion of the solution). To check valve performance, we observed the response of a rotary biomotor, F1-ATPase, for several buffer solutions in the fabricated chamber. [ABSTRACT FROM AUTHOR]

Nivolumab was developed as a monoclonal antibody against programmed death receptor-1, an immune checkpoint inhibitor which negatively regulates T-cell proliferation and activation. Intravenous administration of nivolumab was approved for the treatment of unresectable malignant melanoma in 2014 in Japan. When advanced melanoma patients were treated with nivolumab, median overall survival became longer. Overall survival rate was significantly better in nivolumab-treated melanoma patients than dacarbazine-treated melanoma patients. Nivolumab had an acceptable long-term tolerability profile, with 22% of patients experiencing grade 3 or 4 adverse events related to the drug. Therefore, nivolumab can become an alternative therapy for advanced malignant melanoma. [ABSTRACT FROM AUTHOR]

Previous research indicates that relative to younger adults, older adults show a larger decline in long-term memory (LTM) for associations than for the components that make up these associations. The purpose of the present study was to investigate whether we can impair associative memory performance in young adults by reducing their working memory (WM) resources, hence providing potential clues regarding the underlying causes of the associative memory deficit in older adults. With two experiments, we investigated whether we can reduce younger adults’ long-term associative memory using secondary tasks in which either storage or processing WM loads were manipulated, while participants learned name–face pairs and then remembered the names, the faces, and the name–face associations. Results show that reducing either the storage or the processing resources of WM produced performance patterns of an associative long-term memory deficit in young adults. Furthermore, younger adults’ associative memory deficit was a function of their performance on a working memory span task. These results indicate that one potential reason older adults have an associative deficit is a reduction in their WM resources but further research is needed to assess the mechanisms involved in age-related associative memory deficits. [ABSTRACT FROM AUTHOR]

Divided attention (DA) at encoding has been shown to significantly disrupt later memory for the studied information. However, what type of processing gets disrupted during DA remains unresolved. In this study, we assessed the degree to which strategic effortful processes are affected under DA by comparing the effects of DA at encoding under intentional and pure incidental learning instructions. In three experiments, participants studied list of words or word pairs under either full or divided attention. Results of three experiments, which used different methodologies, converged to show that the effects of DA at encoding reduce memory performance to the same degree under incidental and intentional learning. Secondary task performance indicated that encoding under intentional learning instructions was more effortful than under incidental learning instructions. In addition, the results indicated enhanced attention to the initial appearance of the words under both types of learning instructions. Results are interpreted to imply that other processes, rather than only strategic effortful ones, might be affected by DA at encoding. [ABSTRACT FROM AUTHOR]

Herein, we aimed to evaluate the recently proposed risk assessment strategies of a cytochrome P450 (CYP) mediated drug–drug interaction (DDI) according to the European Medicines Evaluation Agency (EMEA) draft guideline, and discuss the differences between this guideline and the Food and Drug Administration (FDA) draft guidance. A retrospective study on reported 35 clinical DDI cases revealed that the EMEA assessment successfully predicts moderate-to-strong DDIs, i.e. drugs that cause more than 2-fold increase in the area under the curve in the presence and absence of CYP inhibitor (AUCi/AUC); however, EMEA tends to overlook weak DDIs with AUCi/AUC ≤ 2 to > 1.25. For CYP3A4 inhibitors, even clinically insignificant DDIs were overemphasized if the intestinal DDI is considered. The differences between unbound fraction in plasma and microsomes account for the discrepancies in DDI risk assessment results between EMEA and FDA assessments. Comparing two assessment results for CYP2D6 and CYP2C9 inhibitors, the FDA assessment suggested potential DDI risks for sulphinpyrazone and amitriptyline, while the EMEA assessment indicated no potential risk for these drugs. Through a retrospective study, we showed practical differences in the DDI assessment strategies of EMEA and FDA and suggested improvements in their current strategies. [ABSTRACT FROM AUTHOR]

The F1-ATPase, the soluble part of FoF1-ATP synthase, is a rotary molecular motor consisting of α3β3γðε. The γ and ε subunits rotate relative to the α3β3ð sub-complex on ATP hydrolysis by the β subunit. The ε subunit is known as an endogenous inhibitor of the ATPase activity of the F1-ATPase and is believed to function as a regulator of the ATP synthase. This inhibition by the ε subunit (ε inhibition) of F1-ATPase from thermophilic Bacillus PS3 was analyzed by single molecule measurements. By using a mutant ε subunit deficient in ATP binding, reversible transitions between active and inactive states were observed. Analysis of pause and rotation durations showed that the ε inhibition takes a different path from the ADP-Mg inhibition. Furthermore, the addition of the mutant ε subunit to the α3β3γ sub-complex was found to facilitate recovery of the ATPase activity from the ADPMg inhibition. Thus, it was concluded that these two inhibitions are essentially exclusive of each other [ABSTRACT FROM AUTHOR]

The article presents a study on the comparison of neurotropism and the function of neuronal cytoskeletal components within the neural propagation of swine hemagglutinating encephalomyelitis virus (HEV) with those of pseudorabies virus (PRV). The comparison of HEV and PRV were carried out by utilizing cultured non-neuronal cells and nerve cells from dorsal root ganglia (DRG) of new-sprung or 2-4 year-old mice. Meanwhile, through the use of cytoskeletal inhibitors, it was intimated that the spread of PRV and HEV among and within nerve cells are dependent upon intermediate filaments and microtubules of nerve cells. Moreover, it indicates that both viruses might be significantly transferred between the axon terminals and cell body of neurons by means of fast axonal flow.

F1-ATPase is a rotary molecular motor driven by ATP hydrolysis that rotates the γ-subunit against the α3β3 ring. The crystal structures of F1, which provide the structural basis for the catalysis mechanism, have shown essentially 1 stable conformational state. In contrast, single-molecule studies have revealed that F1 has 2 stable conformational states: ATP-binding dwell state and catalytic dwell state. Although structural and single-molecule studies are crucial for the understanding of the molecular mechanism of F1, it remains unclear as to which catalytic state the crystal structure represents. To address this issue, we introduced cysteine residues at βE391 and γR84 of F1 from thermophilic Bacillus PS3. In the crystal structures of the mitochondrial F1, the corresponding residues in the ADP-bound β (βDP) and γ were in direct contact. The βE190D mutation was additionally introduced into the β to slow ATP hydrolysis. By incorporating a single copy of the mutant β-subunit, the chimera F1, α3β2β(E190D/E391 C)β(R84C), was prepared. In single-molecule rotation assay, chimera F1 showed a catalytic dwell pause in every turn because of the slowed ATP hydrolysis of β(E190D/E391C). When the mutant β and γ were cross-linked through a disulfide bond between βE391C and γR84C, F1 paused the rotation at the catalytic dwell angle of β(E190D/E391C), indicating that the crystal structure represents the catalytic dwell state and that βDP is the catalytically active form. The former point was again confirmed in experiments where F1 rotation was inhibited by adenosine-5'- (β,γ-imino)-triphosphate and/or azide, the most commonly used inhibitors for the crystallization of F1. [ABSTRACT FROM AUTHOR]

A rotary motor F1, a catalytic part of ATP synthase, makes a 120° step rotation driven by hydrolysis of one ATP, which consists of 80° and 40° substeps initiated by ATP binding and probably by ADP and/or Pi dissociation, respectively. During active rotations, F1 spontaneously fails in ADP release and pauses after a 80° substep, which is called the ADP-inhibited form. In the present work, we found that when pushed > +40° with magnetic tweezers, the pausing F1 resumes its active rotation after releasing inhibitory ADP. The rate constant of the mechanical activation exponentially increased with the pushed angle, implying that F1 weakens the affinity of its catalytic site for ADP as the angle goes forward. This finding explains not only its unidirectional nature of rotation, but also its physiological function in ATP synthesis; it would readily bind ADP from solution when rotated backward by an F0 motor in the ATP synthase. Furthermore, the mechanical work for the forced rotation was efficiently converted into work for expelling ADP from the catalytic site, supporting the tight coupling between the rotation and catalytic event. [ABSTRACT FROM AUTHOR]

Astaxanthin (3,3′-dihydroxy-β,β-carotene-4,4′-dione) acts as a protective agent against oxidative damage to cells and is widely used in various industries. In this study, we profiled fatty acids in the astaxanthin-producing heterobasidomycete Xanthophyllomyces dendrorhous. Our analysis revealed that X. dendrorhous can produce α-linolenic acid (C18:3ω3) in its cells. To identify plant oils with positive effects on X. dendrorhous , we screened various oils and tested for coproduction of astaxanthin and fatty acids in the yeast. Interestingly, culturing X. dendrorhous with 0.01% (w/v) meadowfoam oil led to a three-fold increase in the growth of the yeast. Due to this increased growth, X. dendrorhous produced more total fatty acids (2.7-fold increase) and astaxanthin (1.4-fold increase) than were produced without oil supplementation. However, fatty acid composition did not change substantially under supplemented conditions. Thus, X. dendrorhous can coproduce industrially valuable astaxanthin and fatty acids, and meadowfoam oil can serve as a promoting factor for this coproduction. • Xanthophyllomyces dendrorhous is able to produce functional fatty acids including α-linolenic acid. • Meadowfoam oil substantially increases the growth of X. dendrorhous. • The increased growth led to improvement of astaxanthin and total fatty acid production in X. dendrorhous. [ABSTRACT FROM AUTHOR]

The valuable marine carotenoid, astaxanthin, is used in supplements, medicines and cosmetics. In this study, crustacyanin, an astaxanthin-binding protein, was used to solubilize and concentrate astaxanthin. The recombinant crustacyanin of European lobster spontaneously formed an inclusion body when it was over-expressed in Escherichia coli. In this study, fusing the NusA-tag to the crustacyanin subunits made it possible to express in a soluble fraction and solubilize astaxanthin in aqueous solution. By cutting off the NusA-tag, the crustacyanin subunits generated the pure insoluble form, and captured and concentrated astaxanthin. Overall, the attaching and releasing NusA-tag method has the potential to supply solubilized carotenoids in aqueous solution and concentrated carotenoids, respectively. [ABSTRACT FROM AUTHOR]

Carotenoids are used commercially for dietary supplements, cosmetics, and pharmaceuticals because of their antioxidant activity. In this study, colored microorganisms were isolated from deep sea sediment that had been collected from Suruga Bay, Shizuoka, Japan. One strain was found to be a pure yellow carotenoid producer, and the strain was identified as Sphingomonas sp. (Proteobacteria) by 16S rRNA gene sequence analysis; members of this genus are commonly isolated from air, the human body, and marine environments. The carotenoid was identified as nostoxanthin ((2,3,2′,3′)-β,β-carotene-2,3,2′,3′-tetrol) by mass spectrometry (MS), MS/MS, and ultraviolet-visible absorption spectroscopy (UV-Vis). Nostoxanthin is a poly-hydroxy yellow carotenoid isolated from some photosynthetic bacteria, including some species of Cyanobacteria. The strain Sphingomonas sp. SG73 produced highly pure nostoxanthin of approximately 97% (area%) of the total carotenoid production, and the strain was halophilic and tolerant to 1.5-fold higher salt concentration as compared with seawater. When grown in 1.8% artificial sea salt, nostoxanthin production increased by 2.5-fold as compared with production without artificial sea salt. These results indicate that Sphingomonas sp. SG73 is an efficient producer of nostoxanthin, and the strain is ideal for carotenoid production using marine water because of its compatibility with sea salt. [ABSTRACT FROM AUTHOR]

The article presents a case study of a 66-year-old female patient who suffered from tinea capitis due to Microsporum canis.

The histologic characteristics of a salivary mucocele in a beagle used in a toxicity study are described in this report. A pale yellowish cyst under the mandibular skin containing frothy mucus was observed at necropsy. Microscopically, numerous villous projections arose from the internal surface of the cyst and were lined by stratified epithelial-like macrophages, which were immunopositive for macrophage scavenger receptor A. A ruptured sublingual interlobar duct connected to the lumen was observed near the cyst. Luminal amorphous material showed a positive reaction with Alcian blue and periodic acid-Schiff staining as did mucin in the sublingual gland. Ultrastructurally, the epithelial-like macrophages had numerous vacuoles containing electron-lucent material, which was presumed to be lysosomal in origin, and had pseudopods on their cell surfaces interdigitating with those on the adjacent cells. This case report helps to understand the diversity of the background findings in beagles used in toxicity studies. [ABSTRACT FROM AUTHOR]

The synuclein family includes three isoforms, termed α, β and γ. α-Synuclein accumulates in various pathological lesions resulting from neurodegenerative disorders including Parkinson''s disease (PD), dementia with Lewy bodies (DLB) and multiple system atrophy. However, neither β- nor γ-synuclein has been detected in Lewy bodies, and thus it is unclear whether these isoforms contribute to neurological pathology. In the present study, we used immunohistochemistry to demonstrate accelerated accumulation of β- and γ-synucleins in axonal spheroids in gracile axonal dystrophy (gad) mice, which do not express ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1). γ-Synuclein immunoreactivity in the spheroids appeared in the gracile nucleus at 3 weeks of age and was maintained until 32 weeks. β-Synuclein immunoreactivity appeared in spheroids around 12 weeks of age. In contrast, α-synuclein immunoreactivity was barely detectable in spheroids. Immunoreactivity for synaptophysin and ubiquitin were either faint or undetectable in spheroids. Given that UCH-L1 deficiency results in axonal degeneration and spheroid formation, our findings suggest that β- and γ-synuclein participate in the pathogenesis of axonal swelling in gad mice. [Copyright &y& Elsevier]

Ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) is a neuron-specific ubiquitin recycling enzyme. A mutation at residue 93 and polymorphism at residue 18 within human UCH-L1 are linked to familial Parkinson’s disease and a decreased Parkinson’s disease risk, respectively. Thus, we constructed recombinant human UCH-L1 variants and examined their structure (using circular dichroism) and hydrolase activities. We confirmed that an I93M substitution results in a decrease in kcat (45.6%) coincident with an alteration in α-helical content. These changes may contribute to the pathogenesis of Parkinson’s disease. In contrast, an S18Y substitution results in an increase in kcat (112.6%) without altering the circular dichroistic spectrum. These data suggest that UCH-L1 hydrolase activity may be inversely correlated with Parkinson’s disease risk and that the hydrolase activity is protective against the disease. Furthermore, we found that oxidation of UCH-L1 by 4-hydroxynonenal, a candidate for endogenous mediator of oxidative stress-induced neuronal cell death, results in a loss of hydrolase activity. Taken together, these results suggest that further studies of altered UCH-L1 hydrolase function may provide new insights into a possible common pathogenic mechanism between familial and sporadic Parkinson’s disease. [Copyright &y& Elsevier]