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BRCA1 maintains genome integrity and suppresses tumorigenesis by promoting homologous recombination (HR)–mediated repair of DNA double-strand breaks (DSB) and DNA damage-induced cell-cycle checkpoints. Phosphorylation of BRCA1 by ATM, ATR, CHK2, CDK, and PLK1 kinases has been reported to regulate its functions. Here we show that ATR and ATM-mediated phosphorylation of BRCA1 on T1394, a highly conserved but functionally uncharacterized site, is a key modification for its function in the DNA damage response (DDR). Following DNA damage, T1394 phosphorylation ensured faithful repair of DSBs by promoting HR and preventing single-strand annealing, a deletion-generating repair process. BRCA1 T1394 phosphorylation further safeguarded chromosomal integrity by maintaining the G2–M checkpoint. Moreover, multiple patient-derived BRCA1 variants of unknown significance were shown to affect T1394 phosphorylation. These results establish an important regulatory mechanism of BRCA1 function in the DDR and may have implications in the development or prognosis of BRCA1-associated cancers. Significance: This study identifies a BRCA1 phosphorylation event critical for its DNA repair function and reveals the functional defects of several BRCA1 variants of unknown significance.

BACKGROUND Several previous studies have reported an unusual root formation in which a fractured apical fragment of an immature root continued to develop independent of the main root after trauma to an immature tooth. To date, there have been only rare reports of the continuing apical formation of the fractured root associated with dens evaginatus (DE). This paper presents a case of a separated root tip formation associated with a fractured tubercle of DE. CASE SUMMARY An 11-year-old boy was referred for gingival sinus on the buccal side of the right mandibular second premolar (tooth # 45). Clinically, tooth # 45 was free of caries, but there was a sign of a fractured tubercle of DE on the occlusal surface. Radiography showed that the root canal of tooth # 45 was widely radiolucent. A separated root apex was found apically under the main root and was nearly completely formed with an apical orifice at the apical tip. Tooth # 45 was diagnosed as tubular fracture of DE with chronic apical periodontitis. A revascularization technique was recommended to treat the tooth. At 3-mo and 1-yr follow-up, the patient remained asymptomatic. Periapical radiography revealed that the separated root tip distally drifted with closure of the apex. However, the root length and thickness of the main root did not increased. CONCLUSION Clinicians should be aware that even if tubercle of DE is fractured in an immature tooth, the root tip may be separated from the main root and completely formed.

In this paper, we develop a general Bayesian hierarchical model for bridging across patient subgroups in phase I oncology trials, for which preliminary information about the dose–toxicity relationship can be drawn from animal studies. Parameters that re-scale the doses to adjust for intrinsic differences in toxicity, either between animals and humans or between human subgroups, are introduced to each dose–toxicity model. Appropriate priors are specified for these scaling parameters, which capture the magnitude of uncertainty surrounding the animal-to-human translation and bridging assumption. After mapping data onto a common, ‘average’ human dosing scale, human dose–toxicity parameters are assumed to be exchangeable either with the standardised, animal study-specific parameters, or between themselves across human subgroups. Random-effects distributions are distinguished by different covariance matrices that reflect the between-study heterogeneity in animals and humans. Possibility of non-exchangeability is allowed to avoid inferences for extreme subgroups being overly influenced by their complementary data. We illustrate the proposed approach with hypothetical examples, and use simulation to compare the operating characteristics of trials analysed using our Bayesian model with several alternatives. Numerical results show that the proposed approach yields robust inferences, even when data from multiple sources are inconsistent and/or the bridging assumptions are incorrect.

It remains controversial to claim blood pressure (BP) as a leading risk factor for high risk of death in peritoneal dialysis patients, and less is known about the relationship between BP and mortality in Chinese peritoneal dialysis patients.From Zhejiang Renal Data System in China, we collected data on patients treated and followed up at 98 peritoneal dialysis centres from 2008 to 2016. The associations of BP parameters [SBP, DBP, mean arterial pressure (MAP) and pulse pressure (PP)] with all-cause and cardiovascular mortality were examined. We fitted Cox models for mortality with penalized splines using nonparametric smoothers. Several sensitivity analyses were performed to confirm the robustness of our primary findings.A total of 7335 Chinese peritoneal dialysis patients were included. During a median follow-up of 35.8 months, 1281 (17.5%) patients died. SBP, DBP, MAP follow a U-shaped pattern of both all-cause and cardiovascular mortality. PP presents a reverse L-shaped association with all-cause mortality. Either a higher (SBP141, DBP85 or MAP102 mmHg) or lower (SBP119, DBP67 or MAP88 mmHg) BP tends to have a significantly higher all-cause and cardiovascular mortality risk. Higher PP (60 mmHg) is related to a higher risk of all-cause mortality, but not cardiovascular mortality. These associations remain the same in our competing risk analysis and subgroup analyses.These data indicate U-shaped associations of SBP, DBP and MAP with all-cause mortality and cardiovascular mortality, respectively, and a reverse L-shaped association of PP with all-cause mortality. Further studies are needed to reliably establish the optimal BP targets for better hypertension control in peritoneal dialysis patients.

Leveraging preclinical animal data for a phase I first-in-man trial is appealing yet challenging. A prior based on animal data may place large probability mass on values of the dose-toxicity model parameter(s), which appear infeasible in light of data accrued from the ongoing phase I clinical trial. In this paper, we seek to use animal data to improve decision making in a model-based dose-escalation procedure for phase I oncology trials. Specifically, animal data are incorporated via a robust mixture prior for the parameters of the dose-toxicity relationship. This prior changes dynamically as the trial progresses. After completion of treatment for each cohort, the weight allocated to the informative component, obtained based on animal data alone, is updated using a decision-theoretic approach to assess the commensurability of the animal data with the human toxicity data observed thus far. In particular, we measure commensurability as a function of the utility of optimal prior predictions for the human responses (toxicity or no toxicity) on each administered dose. The proposed methodology is illustrated through several examples and an extensive simulation study. Results show that our proposal can address difficulties in coping with prior-data conflict commencing in sequential trials with a small sample size., 26 pages, 6 figures; accepted by Biometrical Journal

Graphical Abstract Demineralization of hard tooth tissues leads to dental caries, which cause health problems and economic burdens throughout the world. A biomimetic mineralization strategy is expected to reverse early dental caries. Commercially available anti-carious mineralizing products lead to inconclusive clinical results because they cannot continuously replenish the required calcium and phosphate resources. Herein, we prepared a mineralizing film consisting of hydroxypropylmethylcellulose (HPMC) and polyaspartic acid-stabilized amorphous calcium phosphate (PAsp-ACP) nanoparticles. HPMC which contains multiple hydroxyl groups is a film-forming material that can be desiccated to form a dry film. In a moist environment, this film gradually changes into a gel. HPMC was used as the carrier of PAsp-ACP nanoparticles to deliver biomimetic mineralization. Our results indicated that the hydroxyl and methoxyl groups of HPMC could assist the stability of PAsp-ACP nanoparticles and maintain their biomimetic mineralization activity. The results further demonstrated that the bioinspired mineralizing film induced the early mineralization of demineralized dentin after 24 h with increasing mineralization of the whole demineralized dentin (3–4 µm) after 72–96 h. Furthermore, these results were achieved without any cytotoxicity or mucosa irritation. Therefore, this mineralizing film shows promise for use in preventive dentistry due to its efficient mineralization capability. Supplementary Information The online version contains supplementary material available at 10.1186/s12951-021-01133-7.

Fasting induces profound changes in the hypothalamic–pituitary–thyroid (HPT) axis. After binding thyroid hormone (TH), the TH receptor beta 2 isoform (THRB2) represses Trh and Tsh subunit genes and is the principle negative regulator of the HPT axis. Using mass spectrometry, we identified a major phosphorylation site in the AF-1 domain of THRB2 (serine 101, S101), which is conserved among many members of the nuclear hormone receptor superfamily. More than 50% of THRB2 is phosphorylated at S101 in cultured thyrotrophs (TαT1.1) and in the mouse pituitary. All other THR isoforms lack this site and exhibit limited overall levels of phosphorylation. To determine the importance of THRB2 S101 phosphorylation, we used the TαT1.1 cell line and S101A mutant knock-in mice (Thrb2(S101A)). We found that TH promoted S101 THRB2 phosphorylation and was essential for repression of the axis at physiologic TH concentrations. In mice, THRB2 phosphorylation was also increased by fasting and mimicked Trh and Tshb repression by TH. In vitro studies demonstrated that a master metabolic sensor, AMP-activated kinase (AMPK) induced phosphorylation at the same site and caused Tshb repression independent of TH. Furthermore, we identified cyclin-dependent kinase 2 (CDK2) as a direct kinase phosphorylating THRB2 S101 and propose that AMPK or TH increase S101 phosphorylation through the activity of CDK2. This study provides a physiologically relevant function for THR phosphorylation, which permits nutritional deprivation and TH to use a common mechanism for acute suppression of the HPT axis.

Background: Most of data available in the literature reported the sperm retrieval rate (SRR) and ICSI results of microdissection testicular sperm extraction (micro-TESE) in non-obstructive azoospermia (NOA) patients with different etiologies. Unfortunately, there is currently a lack of comprehensive Intent-to-treat (ITT) data to guide clinicians in conducting comprehensive consultations with NOA patients. The aim of current study was to obtain more comprehensive evidence-based data and clinical outcomes for better consultation of NOA patients who opted to undergo micro-TESE combined with ICSI-IVF.Materials and methods: A retrospective study involved 968 NOA patients underwent micro-TESE during the period between January 2015 and December 2019. Those who had successful sperm retrieval and performed ICSI-IVF cycles were included in intent-to-treat (ITT) analysis. The primary outcome measure was the live birth rate (LBR). The cumulative pregnancy or live birth was defined as clinical pregnancy or at least one live-born baby resulting from an ICSI initiated cycle. Two kinds of stratification analyses were performed based on different etiologies of NOA and various amounts of sperm retrieved. The 'Student's t-test was used for comparison of continuous variables. One-way ANOVA was used to assess outcomes among more than two groups. Chi-squared (χ2) or Fisher's exact test was used for proportions.Results: The SRR of all 968 NOA patients undergoing micro-TESE was 44.6% (n=432). ITT analysis was performed in 424 patients, and ICSI-IVF was applied in 362 couples, leading to 171 clinical pregnancies (40.3%) and 161 live-birth deliveries (38.0%) in the first embryo transfer cycle. No significant difference was observed for per-protocol analysis between the groups of frozen sperm and fresh sperm in cumulative clinical pregnancy rate (CPR, 51.0% vs. 45.2%) and live-birth rate (LBR, 47.5% vs. 42.9%). NOA patients with Y chromosome azoospermia factor c (AZFc) microdeletions had the lowest rate of the high-score embryo on day 3 (4.4%, PPvs. 49.2%, PConclusions: Micro-TESE is an effective sperm retrieval technique for NOA patients. Our data indicated no significant difference in the LBR between ICSI-IVF cycles using frozen or fresh testicular sperm.

BackgroundDespite progress that has been made in the treatment of many immune-mediated inflammatory diseases (IMIDs), there remains a need for improved treatments. Randomised controlled trials (RCTs) provide the highest form of evidence on the effectiveness of a potential new treatment regimen, but they are extremely expensive and time consuming to conduct. Consequently, much focus has been given in recent years to innovative design and analysis methods that could improve the efficiency of RCTs. In this article, we review the current use and future potential of these methods within the context of IMID trials.MethodsWe provide a review of several innovative methods that would provide utility in IMID research. These include novel study designs (adaptive trials, Sequential Multi-Assignment Randomised Trials, basket, and umbrella trials) and data analysis methodologies (augmented analyses of composite responder endpoints, using high-dimensional biomarker information to stratify patients, and emulation of RCTs from routinely collected data). IMID trials are now well-placed to embrace innovative methods. For example, well-developed statistical frameworks for adaptive trial design are ready for implementation, whilst the growing availability of historical datasets makes the use of Bayesian methods particularly applicable.To assess whether and how these innovative methods have been used in practice, we conducted a review via PubMed of clinical trials pertaining to any of 51 IMIDs that were published between 2018 and 20 in five high impact factor clinical journals.ResultsAmongst 97 articles included in the review, 19 (19.6%) used an innovative design method, but most of these were relatively straightforward examples of innovative approaches. Only two (2.1%) reported the use of evidence from routinely collected data, cohorts, or biobanks. Eight (9.2%) collected high-dimensional data.ConclusionsApplication of innovative statistical methodology to IMID trials has the potential to greatly improve efficiency, to generalise and extrapolate trial results, and to further personalise treatment strategies. Currently, such methods are infrequently utilised in practice. New research is required to ensure that IMID trials can benefit from the most suitable methods.

Purpose To investigate the temporal characteristics of visual processing at the fovea and the periphery in high myopia. Methods Eighteen low (LM, ≤ -0.50 and > -6.00 D) and 18 high myopic (HM, ≤ -6.00 D) participants took part in this study. The contrast thresholds in an orientation discrimination task under various stimulus onset asynchrony (SOA) masking conditions were measured at the fovea and a more peripheral area (7°) for the two groups. An elaborated perceptual template model (ePTM) was fit to the behavioral data for each participant. Results An analysis of variance with three factors (SOA, degree of myopia and eccentricity) was performed on the threshold data. The interaction between SOA and degree of myopia in the fovea was significant (F (4, 128) = 2.66, P = 0.036), suggesting that the masking effect had different temporal patterns between the two groups. The temporal profiles for the two groups were derived based on the ePTM model. The peak and the spread of the temporal window in the fovea were much lower and wider, respectively, in the HM group than that in the LM group (both Ps < 0.05). There was no significant difference in the peripheral temporal window between the two groups. Conclusions High myopia is associated with defective temporal processing in the fovea, captured by a flattened temporal window.

Dentin hypersensitivity is attributable to the exposed dentin and its patent tubules. We proposed the therapeutic management of demineralized dentin surfaces using a mineralizing adhesive to seal and remineralize dentin, dentinal tubules, and odontoblast processes. An experimental self-etch adhesive and a mineralizing adhesive consisting of the self-etch adhesive and 20 wt % poly-aspartic acid-stabilized amorphous calcium phosphate (PAsp-ACP) nanoparticles were prepared and characterized by X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, transmission electron microscopy (TEM), and scanning electron microscopy. After 60 acid-etched midcoronal dentin disks were treated with distilled water (control), a desensitizing agent (Gluma), the experimental self-etch adhesive, and the mineralizing adhesive, dentin permeability was measured and mineralization was evaluated by Raman, FTIR, XRD, TEM, and selected-area electron diffraction, irrespective of abrasive and acidic challenges. In vitro cytotoxicity of the adhesive and the mineralizing adhesive was assessed by Cell Counting Kit-8. The mineralizing adhesive possessed excellent biocompatibility. We proposed a hybrid mineralization layer composed of the light-cured mineralizing adhesive and the mineralized dentin surfaces, as well as interiorly mineralized resin tags and odontoblast processes inside of the dentinal tubules. This hybrid mineralization not only reduced dentin permeability but also resisted abrasive and acidic attacks.

The phytohormone auxin controls a myriad of processes in plants, at least in part through its regulation of cell expansion. The "acid growth hypothesis" has been proposed to explain auxin-stimulated cell expansion for five decades, but the mechanism underlying auxin-induced cell wall acidification is poorly characterized. Auxin induces the phosphorylation and activation of the plasma membrane (PM) H+-ATPase that pumps protons into the apoplast, yet how auxin activates its phosphorylation remains elusive. Here, we show that the transmembrane kinase (TMK) auxin signaling proteins interact with PM H+-ATPases and activate their phosphorylation to promote cell wall acidification and hypocotyl cell elongation in Arabidopsis. Auxin induced TMK's interaction with H+-ATPase on the plasma membrane within 1-2 minutes as well as TMK-dependent phosphorylation of the penultimate Thr residue. Genetic, biochemical, and molecular evidence demonstrates that TMKs are required for auxin-induced PM H+-ATPase activation, apoplastic acidification, and cell expansion. Thus, our findings reveal a crucial connection between auxin and PM H+-ATPase activation in regulating apoplastic pH changes and cell expansion via TMK-based cell surface auxin signaling.

The phytohormone auxin controls many processes in plants, at least in part through its regulation of cell expansion1. The acid growth hypothesis has been proposed to explain auxin-stimulated cell expansion for five decades, but the mechanism that underlies auxin-induced cell-wall acidification is poorly characterized. Auxin induces the phosphorylation and activation of the plasma membrane H+-ATPase that pumps protons into the apoplast2, yet how auxin activates its phosphorylation remains unclear. Here we show that the transmembrane kinase (TMK) auxin-signalling proteins interact with plasma membrane H+-ATPases, inducing their phosphorylation, and thereby promoting cell-wall acidification and hypocotyl cell elongation in Arabidopsis. Auxin induced interactions between TMKs and H+-ATPases in the plasma membrane within seconds, as well as TMK-dependent phosphorylation of the penultimate threonine residue on the H+-ATPases. Our genetic, biochemical and molecular evidence demonstrates that TMKs directly phosphorylate plasma membrane H+-ATPase and are required for auxin-induced H+-ATPase activation, apoplastic acidification and cell expansion. Thus, our findings reveal a crucial connection between auxin and plasma membrane H+-ATPase activation in regulating apoplastic pH changes and cell expansion through TMK-based cell surface auxin signalling., Auxin induces transmembrane-kinase-dependent activation of H+-ATPase in the plasma membrane through phosphorylation of its penultimate threonine residue, promoting apoplastic acidification and hypocotyl cell elongation in Arabidopsis.

Triple-negative breast cancer (TNBC) is highly aggressive, difficult to treat and commonly develops visceral metastasis, including lung metastasis. We observed that High mobility group box 1 protein (HMGB1) was highly expressed in human TNBC and positively correlated with cancer metastasis. The hypoxic tumor environment is known to regulate HMGB1 secretion, but an understanding of the underlying mechanism by which tumor-derived HMGB1 regulates interstitial components and promotes breast cancer lung metastasis has remained elusive. The results of the present study showed that the number of CD62Ldim neutrophils, which have a strong ability to produce neutrophil extracellular traps (NETs), increased significantly in both peripheral blood and lung tissues in a mouse TNBC model and were regulated by tumor-derived HMGB1 through the TLR2 pathway. Furthermore, serum HMGB1 levels were positively correlated with CD62Ldim neutrophils in 86 breast cancer patients. We demonstrated that CD62Ldim neutrophils accelerated lung metastasis and that interventions targeting the “HMGB1-CD62Ldim neutrophil-NETs” axis could inhibit lung metastasis. Our results suggest that the combination of HMGB1 and CD62Ldim neutrophils is a potential marker for breast cancer lung metastasis and is novel target for future prevention and therapy.

Purpose Aging affects a variety of visual functions. In this study, we aim to quantitatively investigate the temporal characteristics of visual processing in aging. Methods Twelve younger (24.1 ± 1.6 years) and 12 older observers (58.4 ± 3.6 years) participated in the study. All participants had normal or corrected-to-normal vision. The contrast thresholds of the participants were measured using an orientation discrimination task with white external noise masks. The target-mask stimulus onset asynchronies were 16.7 ms, 33.4 ms, 50.0 ms, 83.4 ms, and ∞ (no external noise masks) in separate conditions. The signal stimulus was carefully chosen such that it was equally visible for the younger and older participants. An elaborated perceptual template model (ePTM) was fit to the data of each participant. Results Without masks, there was no difference in contrast thresholds between the younger and older groups (P = 0.707). With masks, contrast thresholds in the older group elevated more than those in the younger group, and the pattern of threshold elevation differed in the two groups. The ePTM fitted the data well, with the older observers having lower template gains than the younger observers (P = 3.58 × 10-6). A further analysis of the weight parameters of the temporal window revealed that the older observers had a flatter temporal window than the younger observers (P = 0.025). Conclusions Age-related temporal processing deficits were found in older observers with normal contrast sensitivity to the signal stimuli. The deficits were accounted for by the inferior temporal processing window of the visual system in aging.

Background Controlled ovarian stimulation (COS) has a negative effect on the endometrial receptivity compared with natural menstrual cycle. Whether it’s necessary to postpone the first frozen embryo transfer (FET) following a freeze-all strategy in order to avoid any residual effect on endometrial receptivity consequent to COS was inconclusive. Objective The purpose of this retrospective study was to explore whether the delayed FET improve the live birth rate and neonatal outcomes stratified by COS protocols after a freeze-all strategy. Methods A total of 4404 patients who underwent the first FET cycle were enrolled in this study between April 2014 to December 2017, and were divided into immediate (within the first menstrual cycle following withdrawal bleeding) or delayed FET (waiting for at least one menstrual cycle and the transferred embryos were cryopreserved for less than 6 months). Furthermore, each group was further divided into two subgroups according to COS protocols, and the pregnancy and neonatal outcomes were analyzed between the immediate and delayed FET following the same COS protocol. Results When FET cycles following the same COS protocol, there was no significant difference regarding the rates of live birth, implantation, clinical pregnancy, multiple pregnancy, early miscarriage, premature birth and stillbirth between immediate and delayed FET groups. Similarly, no significant differences were found for the mean gestational age, the mean birth weight, and rates of low birth weight and very low birth weight between the immediate and delayed FET groups. The sex ratio (male/female) and the congenital anomalies rate also did not differ significantly between the two FET groups stratified by COS protocols. Conclusion Regardless of COS protocols, FET could be performed immediately after a freeze-all strategy for delaying FET failed to improve reproductive and neonatal outcomes.

Oxytocin and vasopressin are neurohypophyseal hormones with sequence similarity and play a central role in bodily homeostatic regulation. Pain is currently understood to be an important phenotype that those two neurohormones strongly downregulate. Nociceptors, the first component of the ascending neural circuit for pain signals, have constantly been shown to be modulated by those peptides. The nociceptor modulation appears to be critical in pain attenuation, which has led to a gradual increase in scientific interest about their physiological processes and also drawn attention to their translational potentials. This review focused on what are recently understood and stay under investigation in the functional modulation of nociceptors by oxytocin and vasopressin. Effort to produce a nociceptor-specific view could help to construct a more systematic picture of the peripheral pain modulation by oxytocin and vasopressin.

Clinical trials have been conducted for the neuronal ceroid lipofuscinoses (NCLs), a group of neurodegenerative lysosomal diseases that primarily affect children. Whereas clinical rating systems will evaluate long-term efficacy, biomarkers to measure short-term response to treatment would be extremely valuable. To identify candidate biomarkers, we analyzed autopsy brain and matching CSF samples from controls and three genetically distinct NCLs due to deficiencies in palmitoyl protein thioesterase 1 (CLN1 disease), tripeptidyl peptidase 1 (CLN2 disease), and CLN3 protein (CLN3 disease). Proteomic and biochemical methods were used to analyze lysosomal proteins, and, in general, we find that changes in protein expression compared with control were most similar between CLN2 disease and CLN3 disease. This is consistent with previous observations of biochemical similarities between these diseases. We also conducted unbiased proteomic analyses of CSF and brain using isobaric labeling/quantitative mass spectrometry. Significant alterations in protein expression were identified in each NCL, including reduced STXBP1 in CLN1 disease brain. Given the confounding variable of post-mortem changes, additional validation is required, but this study provides a useful starting set of candidate NCL biomarkers for further evaluation.

Vanadium extraction of vanadium-bearing titanomagnetite was investigated by selective chlorination. Thermodynamics analyses on the interactive reactions among related species in the system were made before the experiments. Some fundamental experiments for extracting vanadium by FeClx as chlorinating agent were conducted over the temperature range of 900-1300 K under air or oxygen atmosphere. The results show that vanadium can be extracted by the selective chlorination, using FeClx, based on thermodynamic analysis and experiment. Vanadium extraction ratio first increases with the increase of temperature, and then decreases with the increase of temperature over the range of 900-1300 K under air or oxygen atmosphere. The higher molar ratio of FeCl3 to oxides (n chl:n oxd) reacting with FeCl3, the higher ratio of vanadium extraction. Under oxygen atmosphere, the vanadium extraction ratio is up to 32% at 1100 K for 2 h by using FeCl3 as chlorinating agent.

Accurate knowledge of the intracellular location of proteins is important for numerous areas of biomedical research including assessing fidelity of putative proteinprotein interactions, modeling cellular processes at a system-wide level and investigating metabolic and disease pathways. Many proteins have not been localized, or have been incompletely localized, partly because most studies do not account for entire subcellular distribution. Thus, proteins are frequently assigned to one organelle whereas a significant fraction may reside elsewhere. As a step toward a comprehensive cellular map, we used subcellular fractionation with classic balance sheet analysis and isobaric labeling/quantitative mass spectrometry to assign locations to >6000 rat liver proteins. We provide quantitative data and error estimates describing the distribution of each protein among the eight major cellular compartments: nucleus, mitochondria, lysosomes, peroxisomes, endoplasmic reticulum, Golgi, plasma membrane and cytosol. Accounting for total intracellular distribution improves quality of organelle assignments and assigns proteins with multiple locations. Protein assignments and supporting data are available online through the Prolocate website (http://prolocate.cabm.rutgers. edu). As an example of the utility of this data set, we have used organelle assignments to help analyze whole exome sequencing data from an infant dying at 6 months of age from a suspected neurodegenerative lysosomal storage disorder of unknown etiology. Sequencing data was prioritized using lists of lysosomal proteins comprising wellestablished residents of this organelle as well as novel candidates identified in this study. The latter included copper transporter 1, encoded by SLC31A1, which we localized to both the plasma membrane and lysosome. The patient harbors two predicted loss of function mutations in SLC31A1, suggesting that this may represent a heretofore undescribed recessive lysosomal storage disease gene.

As an indicator of the antioxidant capability of plants, catalase can detoxify reactive oxygen species (ROS) generated by environmental stresses. Sweet potato is one of the top six most important crops in the world. However, its catalases remain largely unknown. In this study, a catalase encoding gene,IbCAT2(accession number: KY615708), was identified and cloned from sweet potato cv. Xushu 18. It contained a 1479 nucleotides’ open reading frame (ORF). S-R-L, Q-K-L, and a putative calmodulin binding domain were located at the C-terminus of IbCAT2, which suggests that IbCAT2 could be a peroxisomal catalase. Next-generation sequencing (NGS) based quantitative analyses showed thatIbCAT2was mainly expressed in young leaves and expanding tuberous roots under normal conditions. When exposed to 10% PEG6000 or 200 mmol/L NaCl solutions,IbCAT2was upregulated rapidly in the first 11 days and then downregulated, although different tissues showed different degree of change. Overexpression ofIbCAT2conferred salt and drought tolerance inEscherichia coliandSaccharomyces cerevisiae. The positive response ofIbCAT2to abiotic stresses suggested thatIbCAT2might play an important role in stress responses.

// Haiyan Zheng 1 , Baohua Yang 2 , Dedong Xu 4 , Wenbo Wang 2 , Jie Tan 3 , Liyuan Sun 3 , Qinghua Li 3 , Li Sun 3 , Xuewei Xia 2, 3 1 Department of Neurosurgery, The Fourth Affiliated Hospital, Zhejiang University School of Medicine, Yiwu, 322000, Zhejiang, China 2 Department of Neurosurgery, Guilin Medical University, Affiliated Hospital, Guilin, 541001, China 3 Guangxi Key Laboratory of Brain and Cognitive Neuroscience, Guilin Medical University, Guilin, 541001, China 4 Department of Neurosurgery, Hainan General Hospital, Haikou, 570311, China Correspondence to: Xuewei Xia, email: xxw7456@163.com Keywords: glioma, immunotherapy, staphylococcal enterotoxin B, interleukin-9, T helper-9 cell Received: June 20, 2016 Accepted: December 05, 2016 Published: December 16, 2016 ABSTRACT The effects of Staphylococcal enterotoxin B (SEB) on regulation of immune response have been recognized; whether SEB can enhance the effects of immunotherapy on glioma remains to be investigated. This study tests a hypothesis that administration with SEB enhances the effects of specific immunotherapy on glioma growth in mice. In this study, a glioma-bearing mouse model was developed by adoptive transfer with GL261 cells (a mouse glioma cell line). The mice were treated with the GL261 cell extracts (used as an Ag) with or without administration of SEB. We observed that treating glioma-bearing mice with the glioma Ag and SEB induced glioma-specific Th9 cells in both glioma tissue and the spleen. Treating CD4 + CD25 − T cells with SEB increased p300 phosphorylation, histone H3K4 acetylation at the interleukin (IL)-9 promoter locus, and increased the IL-9 transcriptional factor binding to the IL-9 promoter. Treating CD4 + CD25 − T cells with both SEB and glioma Ag induced glioma-specific Th9 cells. The glioma-specific Th9 cells induced glioma cell apoptosis in the culture. Treating the glioma-bearing mice with SEB and glioma Ag significantly inhibited the glioma growth. In conclusion, SEB plus glioma Ag immunotherapy inhibits the experimental glioma growth, which may be a novel therapeutic remedy for the treatment of glioma.

Pancreas segmentation is a challenging task in medical image analysis especially for the patients with pancreatic cancer. First, the images often have poor contrast and blurred boundaries. Second, there exist large variations in gray scale, texture, location, shape and size among pancreas images. It becomes even worse with cases of pancreatic cancer. Besides, as an inevitable phenomenon, some of the slices have disconnected topology in pancreas part. All these problems lead to high segmentation uncertainties and make the results inaccurate. Existing pancreas segmentation methods rarely achieve sufficiently accurate and robust results especially for cancer cases. To tackle these problems, we propose a 2D deep learning-based method which can involve uncertainties in the process of segmentation iteratively. The proposed method describes the uncertain regions of pancreatic MRI images based on shadowed sets theory. The results are further corrected through increasing the weights of uncertain regions in iterative training. We evaluate our approach on a challenging pancreatic cancer MRI images dataset collected from the Changhai Hospital, and also validate our approach on the NIH pancreas segmentation dataset. The experimental results demonstrate that our proposed method outperforms the state-of-the-art methods in terms of the Dice similarity coefficient of 73.88% on cancer MRI dataset and 84.37% on NIH dataset respectively.

BACKGROUND The efficacy of an ultrasound guided transmuscular quadratus lumborum block (QLB) for perioperative analgesia of the upper and lower abdomen remain debatable. The purpose of this study was to compare the cutaneous sensory blocked area (CSBA) between QLB blocks performed at the L2 vs. L4 levels. METHODS Twenty-two healthy volunteers were randomized 1:1 to receive an ultrasound guided right transmuscular QLB at the L2 level (group QL2) or L4 level (group QL4). A cold stimulus was applied for testing of the CSBA at 30 minutes after the blockade was performed. The CSBA was mapped and then calculated. Three hours after the QLB, a cold stimulus was applied once every hour until sensation returned normal and the effective block duration for each volunteer was determined and recorded. RESULTS The maximum cephalad dermatome level reached was T7 in group QL2 vs. T11 in group QL4, respectively. Caudally, both groups reached the L2 dermatome level. The QL2 block primarily affected dermatomes T9 to L1, while the QL4 block affected T11 to L1. The total CSBA was larger in QL2 group than that in QL4 group (748 [171] cm2 vs. 501 [186] cm2, P=0.004). The effective duration of the QLB was significantly longer in group QL2 than in group QL4 (18.5 [2.0]h vs. 14.1 [4.7]h, P=0.012). The number of affected dermatomes assessed by cold test was significantly larger for the volunteers in groups QL2 (4.6 [0.81] vs. 2.1 [0.30], P

Treatments are emerging for the neuronal ceroid lipofuscinoses (NCLs), a group of similar but genetically distinct lysosomal storage diseases. Clinical ratings scales measure long-term disease progression and response to treatment but clinically useful biomarkers have yet to be identified in these diseases. We have conducted proteomic analyses of brain and cerebrospinal fluid (CSF) from mouse models of the most frequently diagnosed NCL diseases: CLN1 (infantile NCL), CLN2 (classical late infantile NCL) and CLN3 (juvenile NCL). Samples were obtained at different stages of disease progression and proteins quantified using isobaric labeling. In total, 8303 and 4905 proteins were identified from brain and CSF, respectively. We also conduced label-free analyses of brain proteins that contained the mannose 6-phosphate lysosomal targeting modification. In general, we detect few changes at presymptomatic timepoints but later in disease, we detect multiple proteins whose expression is significantly altered in both brain and CSF of CLN1 and CLN2 animals. Many of these proteins are lysosomal in origin or are markers of neuroinflammation, potentially providing clues to underlying pathogenesis and providing promising candidates for further validation.

Significance Coffee consumption is linked with reduced risk of Parkinson’s disease (PD), and caffeine is generally believed to be the protective agent. However, several lines of evidence suggest the presence of additional compound(s) in coffee that can be protective as well. Here we show that eicosanoyl-5-hydroxytryptamide, which we purified from coffee as an agent that leads to enhanced enzymatic activity of the specific phosphatase PP2A that dephosphorylates the pathogenic protein α-synuclein, works in synergy with caffeine in protecting against mouse models of PD and Dementia with Lewy bodies. The mechanism of this synergy is also through enhancing PP2A, which is dysregulated in the brains of individuals with these α-synucleinopathies., Hyperphosphorylated α-synuclein in Lewy bodies and Lewy neurites is a characteristic neuropathological feature of Parkinson’s disease (PD) and Dementia with Lewy bodies (DLB). The catalytic subunit of the specific phosphatase, protein phosphatase 2A (PP2A) that dephosphorylates α-synuclein, is hypomethylated in these brains, thereby impeding the assembly of the active trimeric holoenzyme and reducing phosphatase activity. This phosphatase deficiency contributes to the accumulation of hyperphosphorylated α-synuclein, which tends to fibrillize more than unmodified α-synuclein. Eicosanoyl-5-hydroxytryptamide (EHT), a fatty acid derivative of serotonin found in coffee, inhibits the PP2A methylesterase so as to maintain PP2A in a highly active methylated state and mitigates the phenotype of α-synuclein transgenic (SynTg) mice. Considering epidemiologic and experimental evidence suggesting protective effects of caffeine in PD, we sought, in the present study, to test whether there is synergy between EHT and caffeine in models of α-synucleinopathy. Coadministration of these two compounds orally for 6 mo at doses that were individually ineffective in SynTg mice and in a striatal α-synuclein preformed fibril inoculation model resulted in reduced accumulation of phosphorylated α-synuclein, preserved neuronal integrity and function, diminished neuroinflammation, and improved behavioral performance. These indices were associated with increased levels of methylated PP2A in brain tissue. A similar profile of greater PP2A methylation and cytoprotection was found in SH-SY5Y cells cotreated with EHT and caffeine, but not with each compound alone. These findings suggest that these two components of coffee have synergistic effects in protecting the brain against α-synuclein−mediated toxicity through maintenance of PP2A in an active state.

Peripheral nociceptors and their synaptic partners utilize neuropeptides for signal transmission. Such communication tunes the excitatory and inhibitory function of nociceptor-based circuits, eventually contributing to pain modulation. Corticotropin-releasing hormone (CRH) is the initiator hormone for the conventional hypothalamic-pituitary-adrenal axis, preparing our body for stress insults. Although knowledge of the expression and functional profiles of CRH and its receptors and the outcomes of their interactions has been actively accumulating for many brain regions, those for nociceptors are still under gradual investigation. Currently, based on the evidence of their expressions in nociceptors and their neighboring components, several hypotheses for possible pain modulations are emerging. Here we overview the historical attention to CRH and its receptors on the peripheral nociception and the recent increases in information regarding their roles in tuning pain signals. We also briefly contemplate the possibility that the stress-response paradigm can be locally intrapolated into intercellular communication that is driven by nociceptor neurons. Such endeavors may contribute to a more precise view of local peptidergic mechanisms of peripheral pain modulation.

Polyphenols are the predominant ingredients in apple seeds. However, few data are available on the phenolic profile or antioxidant activity in apple seeds in previous researches. In this study, low-molecular-weight phenolic compounds and antioxidant activity in seeds, peels, and flesh of seven apple cultivars grown in northwest China were measured and analyzed using HPLC and FRAP, DPPH, ABTS assays, respectively. HPLC analysis revealed phloridzin as the dominant phenolic compound in the seeds with its contents being 240.45–864.42mg/100gDW. Total phenolic content (TPC) measured by the Folin–Ciocalteu assay in apple seed extracts of seven cultivars ranged from 5.74 (Golden Delicious) to 17.44 (Honeycrisp)mgGAE/gDW. Apple seeds showed higher antioxidant activity than peels or flesh; antioxidant activity in seeds varied from 57.59 to 397.70μM Trolox equivalents (TE)/g FW for FRAP, from 37.56 to 64.31μM TE/g FW for DPPH, and from 220.52 to 708.02μM TE/g FW for ABTS. TPC in apple seeds was significantly correlated with all three assays. Principal component analysis (PCA) indicated that Honeycrisp was characterized with high contents of total polyphenols and phloridzin. Our findings suggest that phenolic extracts from apple seeds have good commercial potential as a promising antioxidant for use in food or cosmetics.

Hepatitis B virus (HBV) replicates its genomic DNA via viral DNA polymerase self-primed reverse transcription of a RNA pre-genome in the nucleocapsid assembled by 120 core protein (Cp) dimers. The arginine-rich carboxyl-terminal domain (CTD) of Cp plays an important role in the selective packaging of viral DNA polymerase-pregenomic (pg) RNA complex into nucleocapsid. Previous studies suggested that the CTD is initially phosphorylated at multiple sites to facilitate viral RNA packaging and subsequently dephosphorylated in association with viral DNA synthesis and secretion of DNA-containing virions. However, our recent studies suggested that Cp is hyper-phosphorylated as free dimers and its dephosphorylation is associated with pgRNA encapsidation. Herein, we provide further genetic and biochemical evidence supporting that extensive Cp dephosphorylation does take place during the assembly of pgRNA-containing nucleocapsids, but not empty capsids. Moreover, we found that cellular protein phosphatase 1 (PP1) is required for Cp dephosphorylation and pgRNA packaging. Interestingly, the PP1 catalytic subunits α and β were packaged into pgRNA-containing nucleocapsids, but not empty capsids, and treatment of HBV replicating cells with core protein allosteric modulators (CpAMs) promoted empty capsid assembly and abrogated the encapsidation of PP1 α and β. Our study thus identified PP1 as a host cellular factor that is co-packaged into HBV nucleocapsids, and plays an essential role in selective packaging of the viral DNA-polymerase-pgRNA complex through catalyzing Cp dephosphorylation., Author summary Selective packaging of pregenomic RNA by core protein dimers into nucleocapsid is a key step of HBV replication and is subjected for the regulation by multiple viral and host cellular factors. HBV core protein phosphorylation and dephosphorylation play an essential role in HBV genome replication. However, the cellular kinases and phosphatases responsible for the biochemical events remain elusive. Identification of cellular protein phosphatase 1 as a host cellular factor catalyzing core protein dephosphorylation and facilitating viral pregenomic RNA packaging into nucleocapsids sheds new light on the molecular mechanism of HBV replication and development of therapeutics to cure chronic HBV infection.

Rationale Deep cerebral venous thrombosis (DCVT) is a rare disease, but always results in poor prognosis. Patient concerns We reported a 79-year-old female with coma after traumatic brain injury (TBI). Diagnosis The epidural hematoma was first diagnosed on non-contrast computerized tomography (CT). The hypodense areas in bilateral thalami and basal ganglia on reexamination CT highly indicated the suspicion of DCVT. Finally, the appearance of thrombosis of the vein of Galen on the computed tomography venography (CTA) and digital subtraction angiography (DSA) confirmed the diagnosis. Interventions The patients received surgery to remove the epidural hematoma. After that, she was treated with oral anticoagulation agent (low molecular weight heparin (LMWH), 180 Axal U/kg 24 h) for 4 weeks, shifted by oral warfarin (2.5 mg qd) for 4 weeks. Outcomes The hypodense areas in bilateral thalami and basal ganglia have been largely reversed. At the time of 6 months after surgery, the patient could take care of herself. Lessons If the CT shows hypodense areas in bilateral thalami and basal ganglia, a diagnosis of DCVT should be suspected once the patients could not recover from the treatment of primary diseases.

A high serum HER-2 extracellular domain (sHER-2 ECD) level has a reverse association with tumor behaviors. In this study, a portable platform for the disease biomarker sHER-2 ECD detection has been established using a pressure-based bioassay. The pressure bioassay consists of a monoclonal antibody immobilized on an eight-well strip, the analyte HER-2, and another monoclonal antibody labeled with the Pt nanoparticles (PtNPs), which have the catalytic ability to decompose H2O2 into H2O and O2(g). The increased pressure due to O2(g) generation is measured by a hand-held pressure meter. A total of 34 serum samples were collected to validate the performance of the pressure bioassay. The results showed that the pressure bioassay platform of HER-2 had a dynamic range from 2 to 50 ng/mL with a limit of detection (LOD) of 2 ng/mL, which was consistent with the ELISA result. In the real serum samples, there was a significant correlation between sHER-2 ECD level and several clinicopathological parameters, especially tissue HER-2 status. Furthermore, the sHER-2 ECD level was found to decrease after targeted therapy in a patient with tHER-2 positive. Overall, this bioassay can facilitate breast cancer diagnosis and prognosis in clinical scenarios and resource-limited areas.

Complete replication of the genome is an essential prerequisite for normal cell division, but a variety of factors can block the replisome, triggering replication stress and potentially causing mutation or cell death. The cellular response to replication stress involves recruitment of proteins to stabilize the replication fork and transmit a stress signal to pause the cell cycle and allow fork restart. We find that the ubiquitously expressed DNA damage response factor 53BP1 is required for the normal response to replication stress. Using primary, ex vivo B cells, we showed that a population of 53BP1−/− cells in early S phase is hypersensitive to short-term exposure to three different agents that induce replication stress. 53BP1 localizes to a subset of replication forks following induced replication stress, and an absence of 53BP1 leads to defective ATR-Chk1-p53 signaling and caspase 3-mediated cell death. Nascent replicated DNA additionally undergoes degradation in 53BP1−/− cells. These results show that 53BP1 plays an important role in protecting replication forks during the cellular response to replication stress, in addition to the previously characterized role of 53BP1 in DNA double-strand break repair.

Effect of abnormal GpG methylation in amniotic fluid cells during the second trimester of pregnancy on adverse health risk of offspring was investigated. In total, 237 sets of amniotic fluid cells were collected from patients who received prenatal diagnosis in the Third Affiliated Hospital of Guangzhou Medical University (Guangzhou, China) from April 2010 to October 2011. Among them, 156 sets were from singleton and 81 sets were from twins. H19 gene was amplified by PCR, and the product was purified and pyrosequencing was used to detect the DNA methylation level of GapG. Follow-up records of the birth outcomes of pregnant women's offspring were collected. Positive rate of DNA amplification in 200 cases of amniotic fluid cells was 84.4% (200/237). Average age of singleton pregnancies was higher than that of twins (P0.05). There was no difference in the methylation level of GapG between singleton and twins (P>0.05), but the abnormal methylation rate of GapG1 in twin fetuses was significantly higher than that of singleton (20.3 vs. 3.6%, χ2=8.364, P=0.004). Offspring sex, singleton or twins, mode of delivery, time of pregnancy, and low birth weight showed no significant effect on GapG methylation level of H19 in the second trimester of pregnancy. No offspring deformities were found regardless of the increased or decreased degree of methylation (P>0.05). The number of fetuses born may cause abnormal GapG1 methylation, but no effect of GapG methylation on the adverse health risk of offspring was found.

The objective of this retrospective study was to evaluate clinical outcomes of early cumulus cells removal and early rescue intracytoplasmic sperm injection (ICSI) in high-risk patients of fertilization failure during human in vitro fertilization (IVF).A total of 5,518 patients were enrolled between January 2014 to December 2016. Of which 505 couples performed short insemination with30% fertilization rate were included in short-term IVF group, 102 patients detected total fertilization failure (TFF) were treated with early rescue ICSI (R-ICSI group), and 4911 couples underwent conventional IVF with overnight co-incubation of gametes (traditional IVF group). The clinical outcomes were analyzed among the three groups.The embryo implantation rates (40.34%, 39.78% and 42.42% for traditional, short-term IVF and R-ICSI groups, respectively) were comparable in the three groups. The clinical pregnancy rates among traditional IVF group, short-term IVF group and R-ICSI group were 57.95%, 57.03% and 60.78%, respectively, and the difference among three groups didn't reach significance.The present study indicated that short insemination had no detrimental effects on clinical outcomes in human IVF and could prevent the occurrence of TFF combined with early rescue ICSI for high-risk patients of fertilization failure, which attained acceptable pregnancy outcomes.

Background Hatching is crucial for mammalian embryo implantation, since difficulties during this process can lead to implantation failure, ectopic pregnancy and consequent infertility. Despite years of intensive researches, how internal and external factors affecting embryo hatch are still largely unclear. Methods The effects of parental genetic material and oxygen concentration on hatch process were examined. Fertilized and parthenogenetic mouse preimplantation embryos were cultured in vitro under 5 and 20% oxygen for 120 h. Zona pellucida drilling by Peizo micromanipulation were performed to resemble the breach by sperm penetration. Results Firstly, parthenogenetic embryos had similarly high blastocyst developmental efficiency as fertilized embryos, but significantly higher hatch ratio than fertilized embryos in both O2 concentrations. 5% O2 reduced the hatch rate of fertilized embryos from 58.2 to 23.8%, but increased that of parthenogenetic embryos from 81.2 to 90.8% significantly. Analogously, 5% O2 decreased the ratio of Oct4-positive cells in fertilized blastocysts, whereas increased that in parthenogenetic blastocysts. Additionally, 5% O2 increased the total embryonic cell number in both fertilized and parthegenetic embryos, when compared to 20% O2, and the total cell number of fertilized embryos was also higher than that of parthegenetic embryos, despite O2 concentration. Real-time PCR revealed that the expression of key genes involving in MAPK pathway and superoxide dismutase family might contribute to preimplantation development and consequent blastocyst hatch in vitro. Finally, we showed that fertilized and parthenogenetic embryos have diverse hatch dynamics in vitro, although the zona pellucida integrity is not the main reason for their mechanistic differences. Conclusion Both parental genetic material and O2 concentration, as the representative of intrinsic and extrinsic factors respectively, have significant impacts on mouse preimplantation development and subsequent hatch dynamics, probably by regulating the gene expression involving in MAPK pathway and superoxide dismutase family to control embryonic cell proliferation and allocation of ICM cells. Electronic supplementary material The online version of this article (10.1186/s12958-018-0356-8) contains supplementary material, which is available to authorized users.

Heparanase(HPSE), an endo-β -D-glucuronidase, is found overexpressed in Ovarian cancer (OC). The purpose of our work was to investigate primitively the possible role of HPSE in the development of OC. In this study, RNA interference (RNAi) with a HPSE small hairpin RNAs(HPSE-shRNA) and plasmid with HPSE were used to identify the effects of HPSE on the regulation of malignant behaviors of OC. OV-90 and SKOV3 were selected as a cell model in vitro and in vivo. The results showed that down-regulation of HPSE can significantly inhibit the proliferative and invasive ability of SKOV3 cells, and up-regulation of HPSE in OV-90 cells showed the opposite effects. Compared with the parental OC cells, HPSE silencing cells exhibited attenuated capacities in developing tumor in nude mice, while the growth tumors xenografts derived from these cells were dramatically regressed. In conclusion, our results suggest that HPSE contributes to the proliferation and metastasis of OC and HPSE might be a potent molecular target for OC treatment.

Background: The currently available polysomnography (PSG) equipments and operating personnel are facing increasing pressure, such situation may result in the problem that a large number of obstructive sleep apnea (OSA) patients cannot receive timely diagnosis and treatment, we sought to develop a nomogram quantifying the risk of OSA for a better decision of using PSG, based on the clinical syndromes and the demographic and anthropometric characteristics. Methods: The nomogram was constructed through an ordinal logistic regression procedure. Predictive accuracy and performance characteristics were assessed with the area under the curve (AUC) of the receiver operating characteristics and calibration plots, respectively. Decision curve analyses were applied to assess the net benefit of the nomogram. Results: Among the 401 patients, 73 (18.2%) were diagnosed and grouped as the none OSA (apnea-hypopnea index [AHI]

Chicken egg yolk is a rich source of nutrients providing high quality proteins, vitamins, minerals, carotenoids and antioxidants. Chicken egg yolk, recovered from whole egg within 24 hours post-lay has been utilized as a starting material in the preparation of a dietary supplement that has been demonstrated to lead to gains in muscle mass in a human clinical study. Further, an oil derived from chicken egg yolk has been utilized as a topical agent to treat third degree burn injury. The molecular changes that take place in fertilized, chicken egg yolk during the first 24 hours post-lay are not well understood. By studying how the protein composition of egg yolk varies with fertility status, one can utilize this knowledge to develop egg yolk-based products that have been optimized for specific applications. In this study, a direct quantitative comparison was made between the proteome of fertilized chicken egg yolk and the proteome of unfertilized chicken egg yolk, both maintained at 20 °C and analyzed within 24 hours post-lay. Egg yolk proteins from each fertility state were digested with trypsin, labeled with distinct chemical labels (tandem mass tag reagents) and then combined in a 1 : 1 ratio. A TMT-labeled tryptic digest derived from chicken egg yolk proteins (fertilized and unfertilized) was separated using high-pH/low-pH reverse-phase chromatography and analyzed using mass spectrometry. 225 protein identifications were made from this TMT-labeled tryptic digest based on a minimum of 2 unique peptides observed per protein. 9 proteins increased in abundance in fertilized egg yolk relative to unfertilized egg yolk and 9 proteins decreased in abundance in fertilized egg yolk relative to unfertilized egg yolk. Some proteins that increased in abundance in fertilized egg yolk play an important role in angiogenesis (pleiotrophin, histidine rich glycoprotein) and defense against pathogens (mannose-binding lectin, β-defensin 11, serum amyloid P-component, ovostatin). Based on this study, fertilized chicken egg yolk may be more useful as a starting material relative to unfertilized chicken egg yolk for the purpose of enriching or isolating proteins with pro-angiogenic and anti-microbial properties.

Spermatids undergo the final steps of maturation during spermiogenesis, a process that necessitates extensive rearrangement of organelles such as the mitochondria. Male infertility has been linked to mitochondrial disorder, for example, hypospermatogenesis and asthenozoospermia. However, the mechanisms that regulate mitochondrial dynamics during spermiogenesis remain largely unknown. We found the glycerol kinase (Gyk)-like proteins glycerol kinase-like 1 (Gykl1) and glycerol kinase 2 (Gk2) were specifically localized to the mitochondria in spermatids. Male mice deficient in either Gykl1 or Gk2 were infertile due to dysfunctional spermatozoa, which exhibited unregulated ATP production, disordered mitochondrial sheath formation, abnormal mitochondrial morphology, and defective sperm tail. We demonstrated that the unique C-terminal sequences found in Gykl1 and Gk2 mediated their targeting to the mitochondrial outer membrane. Furthermore, both Gykl1 and Gk2 could interact with Pld6 (MitoPLD) and induce Pld6 and phosphatidic acid (PA)-dependent mitochondrial clustering in cells. Taken together, our study has revealed previously unsuspected functions of Gyk-like proteins in spermiogenesis, providing new insight into the potential mechanisms that lead to spermatozoa dysfunction and male infertility.

Background This study aimed to summarize common pathogens and associated drug resistance in neonatal sepsis (NS). Methods Blood culture and drug sensitivity results from 96 NS cases treated from January 2010 to August 2014 were retrospectively analyzed. Results A total of 97 pathogenic bacteria were detected from these 96 NS cases; Gram-positive cocci accounted for 76.3% of the cases, among which 70.1% involved coagulase-negative staphylococcus (CONS), whereas Gram-negative bacilli and fungi accounted for 19.6% and fungi 4.1% of cases, respectively. Gram-positive cocci exhibited a higher penicillin resistance rate and full vancomycin sensitivity, whereas Gram-negative bacilli exhibited a higher cephalosporin resistance rate, low meropenem resistance rate (6.7%), and no resistance to amikacin. Conclusions The main causative pathogens of NS in our hospital were Gram-positive cocci, among which coagulase-negative Staphylococcus spp such as S. epidermidis and S. haemolyticus were the main conditional pathogens; among Gram-negative pathogens, Klebsiella pneumoniae were most frequently isolated and showed widespread resistance to penicillins and cephalosporins.

The compressive strength of MgO-fluxed pellets was investigated before and after they were reduced. The porosity and pore size of green pellets, product pellets, and reduced pellets were analyzed to clarify how MgO affects the strength of the pellets. Experimental results show that when the MgO-bearing flux content in the pellets increases from 0.0wt% to 2.0wt%, the compressive strength of the pellets at ambient temperature decreases, but the compressive strength of the pellets after reduction increases. Therefore, the compressive strength of the pellets after reduction exhibits no certain positive correlation with that before reduction. The porosity and pore size of all the pellets (with different MgO contents) increase when the pellets are reduced. However, the increase in porosity of the MgO-fluxed pellets is relatively smaller than that of the traditional non-MgO-fluxed pellets, and the pore size range of the MgO-fluxed pellets is relatively narrower. The reduction swelling index (RSI) is a key factor for governing the compressive strength of the reduced pellets. An approximately reversed linear relation can be concluded that the lower the RSI, the greater the compressive strength of the reduced pellets is.

Metformin, one of the most widely prescribed antihyperglycemic drugs, has recently received increasing attention for its potential effects with regard to cancer prevention and treatment. However, the mechanisms behind the suppression of cancer cell growth by metformin remain far from completely understood. The aim of the present study was to investigate whether metformin could regulate histone modification and its downstream gene transcription, and its potential function in inhibiting breast cancer cell proliferation. A T47D cell proliferation curve was determined by cell counting following metformin treatment with differing doses or time courses. The cell cycle was analyzed by flow cytometry with propidium iodide staining. Histone H2B monoubiquitination was evaluated by western blotting subsequent to histone extraction. The histone H2B monoubiquitination downstream gene expression level was determined by quantitative PCR. The results showed that metformin changed the cell-cycle check-point and inhibited breast cancer cell proliferation in a dose-dependent manner. AMPK was activated and histone H2B monoubiquitination and downstream gene transcription were inhibited following metformin treatment in the T47D cells. The effect of metformin on T47D cell proliferation was dependent on AMPK activity. It was concluded that metformin can suppress breast cancer cell growth by the activation of AMPK and the inhibition of histone H2B monoubiquitination and downstream gene transcription. This study reveals a novel potential mechanism of cancer cell growth suppression by metformin.

Calcined magnesite is a binding additive and an MgO-bearing flux for pellets production. The effects of calcination temperature and time on the characteristics of calcined magnesite were investigated. Experimental results indicated that the best calcination condition was 850 °C and 1 h. Under this condition, the hydration activity of the calcined magnesite was 80.56%, and the average diameter of crystal grain D, specific surface area S and the medium particle size D50 were 25.4 nm, 45.40 m2/g and 3.41 μm, respectively. This kind of calcined magnesite was a good binding additive for pellets production. At the same proportion of calcined magnesite, the effects of activities of calcined magnesite on metallurgical properties of green pellet and indurated pellet showed that calcined magnesite with high activity could improve the dropping strength and compressive strength of green pellet and enhance the burst temperature of green pellet; however, the effects of activity on compressive strength, low-temperature reduction degradation index, reduction swelling index and reduction index of indurated pellet were not obvious.

Purpose The contrast sensitivity function (CSF) is measured traditionally with grating stimuli. Recently, we introduced a new set of digit stimuli to improve the efficiency of CSF tests for people unfamiliar with the Latin alphabet. Given the significant differences between grating and digit stimuli, we conducted this study to evaluate whether the estimated CSFs from the digit test are equivalent to those from the grating test. Methods The CSFs of five young (with Psi) and five older (with quick CSF [qCSF]) participants were measured with a two-alternative forced choice (2AFC) grating orientation identification task and a 10-digit identification task. The CSFs obtained from the two tasks were compared. Results The estimated CSFs from the two tasks matched well after controlling for stimulus types and performance levels. The root mean square error (RMSE) between the CSFs from the two tasks was 0.093 ± 0.029 (300 trials) and 0.131 ± 0.016 (100 trials) log10 units for young and older observers, respectively. To reach the same standard deviation (0.1 log10 units), the digit CSF test required fewer trials/less time than the classic grating CSF for young (60 vs. 90 trials) and older (15 vs. 21 trials) observers. The complicated behavioral responses of the observer in the 10-AFC digit identification task can be accounted by a model that consists of digit similarity and one single parameter of sensory noise (χ2[99] = 3.42, P = 0.999). Conclusions The estimated CSFs from the digit test highly matched those obtained from the grating test; however, the digit test is much more efficient. Translational relevance The digit CSF test provides a compatible assessment of the CSF as the traditional grating CSF test with more efficiency.

As a main charging form of BF (blast furnace), pellets play an important role in blast furnace process. However, comparing with sinters, pellets have many disadvantages, such as reduction swelling, low softening and melting temperature and so on. Therefore, the flux pellets have been applied in blast furnace widely, especially MgO containing pellets. The light burned magnesite is applied as MgO containing additive in pellet production. The characters of light burned magnesite are explored. Meanwhile, the effects of it on low-temperature metallurgical properties are investigated such as low-temperature reduction degradation index (RDI), compressive strength (CS) and the reduction swelling index (RSI). The light burned magnesite calcined at 850 °C manifests better grindability, larger specific surface area, and higher hydration activity. It is found that the addition of light burned magnesite can improve low-temperature metallurgical properties (RDI, RSI) of the pellets. With the increase of light burned magnesite in pellets, the RSI and RDI decrease gradually; when the proportion of light burned magnesite does not exceed 2.0% in pellets, the CS decreases slightly, but it still surpasses 2 689 N, which can still meet the demand of BF.