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The SARS-CoV-2 pandemic alerted the potential for significant harm due to future cross-species transmission of various animal coronaviruses to human. There is a significant need of antibody-based drugs to treat patients infected with previously unseen coronaviruses. In this study, we generated CV804, an antibody that binds to the S2 domain of SARS-CoV-2 spike protein, which is highly conserved across the coronavirus family and less susceptible to mutations. CV804 demonstrated broad cross-reactivities not only disease-associated human beta coronaviruses including SARS-CoV, MERS-CoV, HCoV-OC43, HCoV-HKU1 and with existing mutant strains of SARS-CoV-2 and but also with 20 representative animal-origin coronaviruses. CV804 exhibits strong antibody-dependent cellular cytotoxicity (ADCC) to SARS-CoV-2 spike protein expressed on cells in vitro, while completely lacks virus-neutralization activity. In animal models, CV804 suppressed disease progression caused by SARS-CoV-2 infection. Structural studies using HDX-MS combined with reactivity analysis with point mutants of recombinant spike proteins revealed that CV804 binds to a unique conformational epitope within the S2 domain of the spike proteins that is highly conserved among various coronaviruses. Overall, obtained data suggest that the non-neutralizing CV804 antibody recognizes the conformational structure of the spike protein displayed on the surface of infected cells and weakens the viral virulence by supporting the host immune cells' attack through ADCC activity in vivo. The CV804 epitope information revealed in this study is useful for designing pan-corona antibody therapeutics and universal coronavirus vaccines for preparing potential future pandemics. [ABSTRACT FROM AUTHOR]

The importance of treating non‐pulmonary vein (PV) foci is increasing with improvements in the durability of PV isolation. We describe two cases wherein non‐PV foci were induced by mechanical irritation and conventional induction in the same area, which was impossible to induce after ablation, suggesting a relationship between mechanical irritation and induction of non‐PV foci. In a recurrent case of non‐PV foci, atrial fibrillation (AF) was induced only by mechanical irritation. No recurrence has been observed since the ablation of the area. Thus, treatment of the AF induction site with mechanical irritation could be considered an effective therapy for non‐PV foci. [ABSTRACT FROM AUTHOR]

Most autophagy-related genes, or ATG genes, have been identified through studies using budding yeast. Although the functions of the ATG genes are well understood, the contributions of individual genes to non-selective and various types of selective autophagy remain to be fully elucidated. In this study, we quantified the activity of non-selective autophagy, the cytoplasm-to-vacuole targeting (Cvt) pathway, mitophagy, endoplasmic reticulum (ER)-phagy and pexophagy in all Saccharomyces cerevisiae atg mutants. Among the mutants of the core autophagy genes considered essential for autophagy, the atg13 mutant and mutants of the genes involved in the two ubiquitin-like conjugation systems retained residual autophagic functionality. In particular, mutants of the Atg8 ubiquitin-like conjugation system (the Atg8 system) exhibited substantial levels of non-selective autophagy, the Cvt pathway and pexophagy, although mitophagy and ER-phagy were undetectable. Atg8-system mutants also displayed intravacuolar vesicles resembling autophagic bodies, albeit at significantly reduced size and frequency. Thus, our data suggest that membranous sequestration and vacuolar delivery of autophagic cargo can occur in the absence of the Atg8 system. Alongside these findings, the comprehensive analysis conducted here provides valuable datasets for future autophagy research. [ABSTRACT FROM AUTHOR]

The endoplasmic reticulum (ER) undergoes selective autophagy called reticulophagy or ER-phagy. Multiple reticulon- and receptor expression enhancing protein (REEP)-like ER-shaping proteins, including budding yeast Atg40, serve as reticulophagy receptors that stabilize the phagophore on the ER by interacting with phagophore-conjugated Atg8. Additionally, they facilitate phagophore engulfment of the ER by remodeling ER morphology. We reveal that Hva22, a REEP family protein in fission yeast, promotes reticulophagy without Atg8-binding capacity. The role of Hva22 in reticulophagy can be replaced by expressing Atg40 independently of its Atg8-binding ability. Conversely, adding an Atg8-binding sequence to Hva22 enables it to substitute for Atg40 in budding yeast. Thus, the phagophore-stabilizing and ER-shaping activities, both of which Atg40 solely contains, are divided between two separate factors, receptors and Hva22, respectively, in fission yeast. Abbreviations: AIM: Atg8-family interacting motif; Atg: autophagy related; DTT: dithiothreitol; ER: endoplasmic reticulum GFP: green fluorescent protein; NAA: 1-naphthaleneacetic acid; REEP: receptor expression enhancing protein; RFP: red fluorescent protein; UPR: unfolded protein response. [ABSTRACT FROM AUTHOR]

Engineered living cells have been designed to perform a broad variety of functions (8-16) but, with few exceptions, complex computational constructs (such as comparators, bit adders or multiplexers) are difficult [...], Ongoing efforts within synthetic and systems biology have been directed towards the building of artificial computational devices (1) using engineered biological units as basic building blocks (2,3). Such efforts, inspired in the standard design of electronic circuits (4-7), are limited by the difficulties arising from wiring the basic computational units (logic gates) through the appropriate connections, each one to be implemented by a different molecule. Here, we show that there is a logically different form of implementing complex Boolean logic computations that reduces wiring constraints thanks to a redundant distribution of the desired output among engineered cells. A practical implementation is presented using a library of engineered yeast cells, which can be combined in multiple ways. Each construct defines a logic function and combining cells and their connections allow building more complex synthetic devices. As a proof of principle, we have implemented many logic functions by using just a few engineered cells. Of note, small modifications and combination of those cells allowed for implementing more complex circuits such as a multiplexer or a 1-bit adder with carry, showing the great potential for re-utilization of small parts of the circuit. Our results support the approach of using cellular consortia as an efficient way of engineering complex tasks not easily solvable using single-cell implementations.

Muscle disuse induces atrophy through increased reactive oxygen species (ROS) released from damaged mitochondria. Mitophagy, the autophagic degradation of mitochondria, is associated with increased ROS production. However, the mitophagy activity status during disuse‐induced muscle atrophy has been a subject of debate. Here, we developed a new mitophagy reporter mouse line to examine how disuse affected mitophagy activity in skeletal muscles. Mice expressing tandem mCherry‐EGFP proteins on mitochondria were then used to monitor the dynamics of mitophagy activity. The reporter mice demonstrated enhanced mitophagy activity and increased ROS production in atrophic soleus muscles following a 14‐day hindlimb immobilization. Results also showed an increased expression of multiple mitophagy genes, including Bnip3, Bnip3l, and Park2. Our findings thus conclude that disuse enhances mitophagy activity and ROS production in atrophic skeletal muscles and suggests that mitophagy is a potential therapeutic target for disuse‐induced muscle atrophy. [ABSTRACT FROM AUTHOR]

We describe an autopsy case of a 75-year-old female with limited cutaneous systemic sclerosis (lcSSc) and gangrene due to macrovascular involvement. She was diagnosed with lcSSc complicated with pulmonary arterial hypertension and digital ulcers 9 years before admission. She had recurrent and refractory lower limb ulcers (LLUs), and died because of sepsis caused by gangrene infection. Autopsy findings revealed severely thickened arterial walls of the visceral organs, consistent with vascular involvement of SSc. Systemic vascular involvement in lcSSc may progress in patients with LLUs who harbour several risk factors for vascular involvement. [ABSTRACT FROM AUTHOR]

Mitochondria produce the majority of ATP required by cells via oxidative phosphorylation. Therefore, regulation of mitochondrial quality and quantity is important for maintaining cellular activities. Mitophagy, the selective degradation of mitochondria, is thought to contribute to control of mitochondrial quality and quantity. In recent years, the molecular mechanism of mitophagy has been extensively studied in yeast and mammalian cells. In particular, identification of the mitophagy receptor Atg32 has contributed to substantial progress in understanding of mitophagy in yeast. This review summarizes the molecular mechanism of mitophagy in yeast and compares it to the mechanism of mitophagy in mammals. We also discuss the current understanding of mitophagy in plants. [ABSTRACT FROM AUTHOR]

Yeast cells undergo diploid-specific developments such as spore formation via meiosis and pseudohyphal development under certain nutrient-limited conditions. Studies on these aspects require homozygous diploid mutants, which are generally constructed by crossing strains of opposite mating-type with the same genetic mutation. So far, there has been no direct way to generate and select diploids from haploid cells. Here, we developed a method for efficient construction of homozygous diploids using a PGAL1-HO gene (galactose-inducible mating-type switch) and a PSTE18-URA3 gene (counter selection marker for diploids). Diploids are generated by transient induction of the HO endonuclease, which is followed by mating of part of the haploid population. Since the STE18 promoter is repressed in diploids, diploids carrying PSTE18-URA3 can be selected on 5-fluoroorotic acid (5-FOA) plates where the uracil prototrophic haploids cannot grow. To demonstrate that this method is useful for genetic studies, we screened suppressor mutations of the complex colony morphology, strong agar invasion and/or hyper-filamentous growth caused by lack of the Hog1 MAPK in the diploid Σ1278b strain background. Following this approach, we identified 49 suppressor mutations. Those include well-known positive regulator genes for filamentous growth signaling pathways, genes involved in mitochondrial function, DNA damage checkpoint, chromatin remodeling, and cell cycle, and also previously uncharacterized genes. Our results indicate that combinatorial use of the PGAL1-HO and PSTE18-URA3 genes is suitable to efficiently construct and select diploids and that this approach is useful for genetic studies especially when combined with large-scale screening. [ABSTRACT FROM AUTHOR]

The article presents a study which examined the suppression of phenotypes of the kexB disruptant in Aspergillus oryzae and Aspergillus nidulas under hyperosmotic conditions. Suppression mechanism of the oryzapsins and processing of recombinant fusion protein were analyzed. Results show that kexB phenotypes were not suppressed by oryzapsins, but the recombinant fusion proteins were cleaved, and concludes that an alternative processing protease in the Aspergillus fungi is possible.

The article discusses a study which presents novel reporter systems for monitoring activation of the Aspergillus nidulans high-osmolarity glycerol response (AnHOG) pathway. It is stated that AnHOG pathway involved in osmoadaptation. The study found that fludioxonil causes improper activation of HogA mitogen-activated protein kinase in A. nidulans. The results indicate that the reporter activity is regulated via HogA activation.

The article examines the role of SskA ad SrrA response regulators in oxidative stress responses of hyphae and asexual spores in the phosphorelay signaling network of fungus Aspergillus nidulans. According to the authors, expression of the vegetative-stage specific catB catalase gene was severely impaired in both mutants. They add the results of the study indicate the physiological importance of the His-Asp phosphorelay signaling network involving the SskA and SrrA response regulators.

Genome sequencing analyses revealed thatAspergillus nidulanshas orthologous genes to all those of the high-osmolarity glycerol (HOG) response mitogen-activated protein kinase (MAPK) pathway ofSaccharomyces cerevisiae.A. nidulansmutant strains lackingsskA,sskB,pbsB,orhogA, encoding proteins orthologous to the yeast Ssk1p response regulator, Ssk2p/Ssk22p MAPKKKs, Pbs2p MAPKK and Hog1p MAPK, respectively, showed growth inhibition under high osmolarity, and HogA MAPK in these mutants was not phosphorylated under osmotic or oxidative stress. Thus, activation of theA. nidulansHOG (AnHOG) pathway depends solely on the two-component signalling system, and MAPKK activation mechanisms in the AnHOG pathway differ from those in the yeast HOG pathway, where Pbs2p is activated by two branches, Sln1p and Sho1p. Expression ofpbsBcomplemented the high-osmolarity sensitivity of yeastpbs2Δ, and the complementation depended on Ssk2p/Ssk22p, but not on Sho1p. Pbs2p requires its Pro-rich motif for binding to the Src-homology3 (SH3) domain of Sho1p, but PbsB lacks a typical Pro-rich motif. However, a PbsB mutant (PbsB(Pro)) with the yeast Pro-rich motif was activated by the Sho1p branch in yeast. In contrast, HogA insskAΔ expressing PbsB(Pro) was not phosphorylated under osmotic stress, suggesting thatA. nidulansShoA, orthologous to yeast Sho1p, is not involved in osmoresponsive activation of the AnHOG pathway. We also found that besides HogA, PbsB can activate another Hog1p MAPK orthologue, MpkC, inA. nidulans, althoughmpkCis dispensable in osmoadaptation. In this study, we discuss the differences between the AnHOG and the yeast HOG pathways. [ABSTRACT FROM AUTHOR]

Reports on the isolation and functional analysis of a gene, tcsB, by encoding a transmembrane hybrid-type histidine kinase from Aspergillus nidulans. TcsB encoding of an A. nidulans hybrid-type histidine kinase; Strains, media and growth conditions; Molecular cloning and sequencing of tcsB gene.

The article analyzes a histidine protein kinase, NikA, that is found in many fungi, with special reference to responses to potent fungicides, iprodione and fluodioxonil. According to the authors, NikA and two downstream response regulators are crucially implicated in the mode of action of fungicides and that the further downstream HogA-mitogen activated protein kinase cascade is exaggerated abnormally in hyphase by the fungicides in a manner dependent on the NikA-SskA phosphorelay.

Background: Given the popularity of pterional craniotomy, numerous modifications have been made to prevent postoperative deformities. With the advent of titanium plates, fixation has become both simple and excellent. However, titanium plates can cause skin problems, infection, or cause skull growth to fail., Methods: To develop a simple, cost-effective, and esthetically satisfactory fixation method, without the use of non-metallic materials, six young and older patients underwent pterional craniotomy. CranioFix Absorbable clamps were used to fix the bone flap in the frontal and temporal regions such that the frontal part was in close contact with the skull. After fixation, the bone chips and bone dust were placed in the bone gap and fixed with fibrin glue. We measured the computed tomography values of the reconstructed area and thickness of the temporal profiles postoperatively over time., Results: Bone fusion was achieved in all patients by 1 year after surgery. Both the thickness of the temporalis muscle and the thickness of the temporal profile had changed within 2 mm as compared with the preoperative state., Conclusion: Our simple craniotomy technique, gentle tissue handling, and osteoplastic cranioplasty yielded satisfactory esthetic results and rigidness in pterional craniotomy., Competing Interests: There are no conflicts of interest., (Copyright: © 2023 Surgical Neurology International.)

Objective: Endovascular coil embolization for anterior communicating artery (ACoA) and anterior cerebral artery (ACA) aneurysms is associated with high total and near-total occlusion rates, but the complication rate is high. The development of newer endovascular technologies may improve the clinical outcomes. This study investigated the status of endovascular treatment of ACoA and ACA aneurysms by comparing our results with past reports., Methods: Between January 2006 and December 2018, we investigated 50 patients who were followed for 12 months or longer to clarify the outcomes of coil embolization. The outcomes of embolization were evaluated using time-of-flight MRA. The safety was evaluated based on procedure-related complications that affected clinical outcomes., Results: Initial assessments demonstrated complete obliteration in 84% (42 of 50 patients) and a residual neck in 14% (7 of 50 patients). Procedure-related complications developed in 12% (6 of 50 patients). The procedure-related morbidity rate was 2% (1 of 50 patients) and there was no procedure-related death. Recanalization was noted in 14% (7 of 50 patients, median follow-up period, 57 months). The recanalized aneurysms were significantly smaller than the stable aneurysms in maximum size (4.3 mm vs. 5.8 mm; p = 0.017) and height (3.7 mm vs. 4.3 mm; p = 0.035)., Conclusion: We demonstrated the safety and effectiveness of endovascular coil embolization for ACoA and ACA aneurysms. The small size of aneurysms may be related to recanalization., Competing Interests: All authors have no conflicts of interest to declare., (©2022 The Japanese Society for Neuroendovascular Therapy.)

Objective: Transvenous embolization (TVE) is typically used in combination with the residual shunt of transarterial embolization (TAE) for the treatment of direct carotid-cavernous fistulas (direct CCFs). This report is about our additional embolization method using combination therapy., Case Presentation: Five consecutive cases of direct CCF were presented; two were caused by aneurysms and three by head injuries. The treatment for each was started with TAE, with the addition of TVE if a shunt remained. At the time of TVE, a microcatheter positioned in the internal carotid artery passing from the cavernous sinus through the aneurysm neck or fistula was pulled back (pull-back method). It was then placed in the coil mass with TAE, and additional coils were filled. In two cases, the shunt disappeared by using only TAE, whereas it disappeared after being additionally embolized by the pull-back method in the remaining cases. All patients recovered with no postoperative complications., Conclusion: The TAE and TVE combination therapy with the pull-back method could efficiently embolize the residual shunt after TAE., Competing Interests: The authors declare no conflicts of interest., (©2022 The Japanese Society for Neuroendovascular Therapy.)

Many eukaryotic histidine-to-aspartate (His-Asp) phosphorelay systems consist of three types of signal transducers: a His-kinase (HK), a response regulator (RR), and a histidine-containing phosphotransfer intermediate (HPt). In general, the HPt acts as an intermediate between the HK and the RR and is indispensable for inducing appropriate responses to environmental stresses. In a previous study, we attempted but were unable to obtain deletion mutants of the ypdA gene in order to characterize its function in the filamentous fungus Aspergillus nidulans . In the present study, we constructed the C ypdA strain in which ypdA expression is conditionally regulated by the A. nidulans alcA promoter. We constructed C ypdA strains with RR gene disruptions (C ypdA-sskA Δ, C ypdA-srrA Δ, and C ypdA-sskA Δ srrA Δ). Suppression of YpdA induced by ypdA downregulation activated the downstream HogA mitogen-activated protein kinase cascade. YpdA suppression caused severe growth defects and abnormal hyphae, with features such as enhanced septation, a decrease in number of nuclei, nuclear fragmentation, and hypertrophy of vacuoles, both regulated in an SskA-dependent manner. Fludioxonil treatment caused the same cellular responses as ypdA suppression. The growth-inhibitory effects of fludioxonil and the lethality caused by ypdA downregulation may be caused by the same or similar mechanisms and to be dependent on both the SskA and SrrA pathways., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Yoshimi, Hagiwara, Ono, Fukuma, Midorikawa, Furukawa, Fujioka, Mizutani, Sato, Miyazawa, Maruyama, Marui, Yamagata, Nakajima, Tanaka and Abe.)

Mitophagy plays an important role in mitochondrial homeostasis. In yeast, the phosphorylation of the mitophagy receptor Atg32 by casein kinase 2 is essential for mitophagy. This phosphorylation is counteracted by the yeast equivalent of the STRIPAK complex consisting of the PP2A-like protein phosphatase Ppg1 and Far3-7-8-9-10-11 (Far complex), but the underlying mechanism remains elusive. Here we show that two subpopulations of the Far complex reside in the mitochondria and endoplasmic reticulum, respectively, and play distinct roles; the former inhibits mitophagy via Atg32 dephosphorylation, and the latter regulates TORC2 signaling. Ppg1 and Far11 form a subcomplex, and Ppg1 activity is required for the assembling integrity of Ppg1-Far11-Far8. The Far complex preferentially interacts with phosphorylated Atg32, and this interaction is weakened by mitophagy induction. Furthermore, the artificial tethering of Far8 to Atg32 prevents mitophagy. Taken together, the Ppg1-mediated Far complex formation and its dissociation from Atg32 are crucial for mitophagy regulation., Competing Interests: AI, KF, TF, TS, KI, SY, TK No competing interests declared, (© 2020, Innokentev et al.)

Degradation of mitochondria through mitophagy contributes to the maintenance of mitochondrial function. In this study, we identified that Atg43, a mitochondrial outer membrane protein, serves as a mitophagy receptor in the model organism Schizosaccharomyces pombe to promote the selective degradation of mitochondria. Atg43 contains an Atg8-family-interacting motif essential for mitophagy. Forced recruitment of Atg8 to mitochondria restores mitophagy in Atg43-deficient cells, suggesting that Atg43 tethers expanding isolation membranes to mitochondria. We found that the mitochondrial import factors, including the Mim1-Mim2 complex and Tom70, are crucial for mitophagy. Artificial mitochondrial loading of Atg43 bypasses the requirement of the import factors, suggesting that they contribute to mitophagy through Atg43. Atg43 not only maintains growth ability during starvation but also facilitates vegetative growth through its mitophagy-independent function. Thus, Atg43 is a useful model to study the mechanism and physiological roles, as well as the origin and evolution, of mitophagy in eukaryotes., Competing Interests: TF, YE, TS, KF, SY, KI, DK, YY, TK No competing interests declared, (© 2020, Fukuda et al.)

Background: This study was performed to clarify whether gut microbiota obtained from fecal samples could identify the type of diabetes in patients of each gender by using a combination of terminal restriction fragment length polymorphism (T-RFLP) analysis and data mining., Methods: A cross-sectional study was performed at three centers. Fecal samples were collected from 12 Japanese patients with type 1 diabetes mellitus (T1D), 18 patients with type 2 diabetes mellitus (T2D), and 31 subjects without diabetes mellitus (non-DM). Amplification of fecal 16S rRNA was carried out. After digestion of the amplification products with restriction enzymes ( Alu I, Bsl I, Hae III, and Msp I), terminal restriction fragments (T-RFs) of DNA were detected. A data mining algorithm (classification and regression tree (CART) modeling system) provides a decision tree that classifies subjects into various groups according to pre-assigned characteristics., Results: Among men, the error rate was 2.4% with Msp I, while error rates were 0.0% with other restriction enzymes. Among women, the error rate was 0.0% with all restriction enzymes. The operational taxonomic units (OTUs) incorporated into the decision tree differed between men and women., Conclusions: We were able to classify the 16SrRNA gene amplification products obtained from fecal samples of T1D patients, T2D patients, and non-DM subjects with a high level of precision by combining T-RFLP analysis and data mining. Specific gut microbiota patterns were found for T1D and T2D patients, as well as a sex difference of the patterns., Competing Interests: The authors declare no conflict of interest.

OBJECTIVE The vascular lumen of an acutely occluded internal carotid artery (ICA) generally contains numerous thrombi. Therefore, carotid angiography on the affected side during revascularization therapy of acute ICA occlusion has a potential risk of causing distal embolization. In this study the authors propose the use of contralateral carotid angiography. METHODS Six patients with acute ICA occlusion underwent revascularization therapy using a stent retriever or Penumbra system. Revascularization therapy was performed with placement of a 9-Fr balloon-guiding catheter (BGC) in the affected ICA and a 4-Fr diagnostic catheter in the contralateral ICA. During the procedure, the 9-Fr BGC was kept inflated, and all control angiography was performed from the 4-Fr diagnostic catheter. After thrombectomy, contralateral carotid angiography combined with manual aspiration from the 9-Fr BGC was performed to assess the presence or absence of residual thrombi in the affected ICA. The 9-Fr BGC was deflated only after the complete absence of residual thrombi in the affected ICA was confirmed. RESULTS The time required for introducing the 4-Fr diagnostic catheter into the contralateral ICA was within a few minutes in all patients. Residual thrombi in the affected ICA were found in 3 of 6 patients. The residual thrombi in these 3 patients were completely removed; thus, distal embolization was prevented. CONCLUSIONS Contralateral carotid angiography is useful for avoiding distal embolization during revascularization therapy of acute ICA occlusion. Further studies involving a larger number of patients are warranted to verify the clinical efficacy of this contralateral carotid angiography.

Mitochondrial autophagy (mitophagy) is a process that selectively degrades mitochondria via autophagy. Recent studies have shown that mitophagy plays an important role in mitochondrial homeostasis by degrading damaged or excess mitochondria. The budding yeast Saccharomyces cerevisiae is a powerful model organism that has been employed to study several biological phenomena. Recently, there has been significant progress in the understanding of mitophagy in yeast following the identification of Atg32, a mitochondrial outer membrane receptor protein for mitophagy. In this chapter, we describe protocols to study mitophagy in yeast via a genome-wide screen for mitophagy-deficient mutants using fluorescence microscopy and immunoblotting.

Background: The demand of a burr hole surgery for chronic subdural hematoma (CSDH) is increasing in the global aging society. Burr hole-derived autologous bone dusts are not associated with extra costs compared with other commonly used synthetic materials. In addition, postoperative calvarium ossification requires periosteum-mediated blood supply, which is lacking after using avascular synthetic materials. Based on these findings, we hypothesized that the combination of the bone plugs and the preserved periosteum during burr hole surgeries for CSDH would induce efficient calvarium ossification., Methods: We evaluated the long-term effects of bone plugs on the degree of ossification and cosmetic appearance of the skin covering the burr hole sites. We included 8 patients (9 burr holes) who received the autologous bone dust derived from burr holes. As the control group, 9 burr holes that did not receive any burr hole plugs were retrospectively selected. These burr holes were evaluated by computed tomography (CT) scan for the calvarium defect ratios, CT value-based ossification, and the degree of skin sinking., Results: Ossification was observed in all the bone plugs by the bone density CT scans; they maintained their volume at 12 months after the surgeries. The calvarium defect ratios (volume ratios of the unossified parts in the burr holes) gradually increased during the first 6 months and reached 0.44 at 12 months. The mean CT values also increased from 527 HU to 750 HU for the first 6 months and reached 905 HU at 12 months. The degrees of skin sinking at the burr hole sites with the bone plugs were 1.24 mm whereas those without the bone plugs were 2.69 mm ( P = 0.004)., Conclusion: Application of burr hole-derived autologous bone dust is associated with better ossification and objective cosmetic result following burr hole surgery after CSDH., Competing Interests: There are no conflicts of interest.

Mitophagy is thought to play an important role in mitochondrial quality control. Mitochondrial division is believed to occur first, and autophagosome formation subsequently occurs to enwrap mitochondria as a process of mitophagy. However, there has not been any temporal analysis of mitochondrial division and autophagosome formation in mitophagy. Therefore, the relationships among these processes remain unclear. We show that the mitochondrial division factor Dnm1 in yeast or Drp1 in mammalian cells is dispensable for mitophagy. Autophagosome formation factors, such as FIP200, ATG14, and WIPIs, were essential for the mitochondrial division for mitophagy. Live-cell imaging showed that isolation membranes formed on the mitochondria. A small portion of the mitochondria then divided from parental mitochondria simultaneously with the extension of isolation membranes and autophagosome formation. These findings suggest the presence of a mitophagy process in which mitochondrial division for mitophagy is accomplished together with autophagosome formation., (© 2016 Yamashita et al.)

Mitogen-activated protein kinases (MAPKs) have a number of targets which they regulate at transcriptional and post-translational levels to mediate specific responses. The yeast Hog1 MAPK is essential for cell survival under hyperosmotic conditions and it plays multiple roles in gene expression, metabolic regulation, signal fidelity and cell cycle regulation. Here we describe essential and non-essential roles of Hog1 using engineered yeast cells in which osmoadaptation was reconstituted in a Hog1-independent manner. We rewired Hog1-dependent osmotic stress-induced gene expression under the control of Fus3/Kss1 MAPKs, which are activated upon osmostress via crosstalk in hog1Δ cells. This approach revealed that osmotic up-regulation of only two Hog1-dependent glycerol biosynthesis genes, GPD1 and GPP2, is sufficient for successful osmoadaptation. Moreover, some of the previously described Hog1-dependent mechanisms appeared to be dispensable for osmoadaptation in the engineered cells. These results suggest that the number of essential MAPK functions may be significantly smaller than anticipated and that knockout approaches may lead to over-interpretation of phenotypic data.

Background: Many diabetes patients who require insulin perform multiple subcutaneous injections every day that often cause pain, discomfort, and anxiety. We compared efficacy (glycemic control) and patient preference for two types of needle: a shorter straight needle (32 gauge×4 mm, straight wall; Nippon Becton Dickinson Co., Ltd., Tokyo, Japan; hereafter referred to as BD32S4) and a thinner microtapered needle (33-gauge tip and 28-gauge base×5 mm, double-tapered wall; Terumo Corp., Tokyo, Japan; hereafter referred to as TR33T5) in a single-center study., Patients and Methods: Eighty-four patients with diabetes were enrolled in a randomized, open-label crossover trial. The patients injected their usual insulin dosage with one type of needle for 4 weeks and then switched to the other type for the next 4 weeks. The serum glycated albumin level was measured before and after each 4-week period. Each patient assessed pain during injection on a 150-mm visual analog scale (VAS). Needle preference, perceptions of handling, and acceptance were assessed by the patients, who completed a questionnaire after using each type of needle for 4 weeks., Results: In total, 79 patients completed the study. There was no difference of glycemic control between the two needles. The mean VAS score was -14.5 mm (95% confidence interval, -20.9, -8.0 mm), indicating that the patients perceived less pain with the BD32S4 needle. In the overall evaluation, a significantly higher percentage of patients selected the BD32S4 as the better needle compared with the TR33T5 (60.3% vs. 19.2%; P<0.0001)., Conclusions: The BD32S4 needle was more highly evaluated and was preferred by the patients with respect to pain during injection, usability, and visual impression, without having a negative impact on glycemic control. The overall preference of patients for the shorter needle in this study suggests that needle length may be one of the major contributing factors for patients' comfort in insulin injection, although the other relevant factors of needles still need to be considered.