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The One-to-one Pickup and Delivery Problem with Shortest-path Transport along Real-life Paths (OPDPSTRP) is presented in this paper. It is a variation of the One-to-one Pickup and Delivery Problem (OPDP), which is common in daily life, such as the Passenger Train Operation Plans (PTOP) and partial Taxi-sharing Problem. Unlike the classical OPDP, in the OPDPSTRP, (1) each demand must be transported along the shortest path according to passengers/shippers requirements, and (2) each vehicle should travel along a real-life path. First, six route structure rules are proposed for the OPDPSTRP, and a kind of Mixed-Integer Programming (MIP) models is formulated for it. Second, A Variable Neighborhood Descent (VND), a Variable Neighborhood Research (VNS), a Multi-Start VND (MS_VND) and a Multi-Start VNS (MS_VNS) with five neighborhood operators has been developed to solve the problem. Finally, The Gurobi solver, the VND, the VNS, the MS_VND and the MS_VNS have been compared with each other by 84 random instances partitioned in small size connected graphs, medium size connected graphs and large size connected graphs. From the test results we found that solutions generated by these approaches are often comparable with those found by the Gurobi solver, and the solutions found by these approaches are better than the solutions found by the Gurobi solver when solving instances with larger numbers of demands. In almost all instances, the MS_VND significantly outperforms the VND and the VNS in terms of solution quality, and outperforms the MS_VNS both in terms of solution quality and CPU time. In the instances with large numbers of demands, the MS_VND is still able to generate good feasible solutions in a reasonable CPU time, which is of vital practical significance for real-life instances.

[This corrects the article DOI: 10.1371/journal.pone.0065381.].

The endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved protein complex involved in inserting the transmembrane domain of membrane proteins into membranes in the ER. EMC3 is an essential component of EMC and is important for rhodopsin synthesis in photoreceptor cells. However, the in vivo function of Emc3 in bipolar cells (BCs) has not been determined. To explore the role of Emc3 in BCs, we generated a BC-specific Emc3 knockout mouse model (named Emc3 cKO) using the Purkinje cell protein 2 (Pcp2) Cre line. Although normal electroretinography (ERG) b-waves were observed in Emc3 cKO mice at 6 months of age, Emc3 cKO mice exhibited reduced b-wave amplitudes at 12 months of age, as determined by scotopic and photopic ERG, and progressive death of BCs, whereas the ERG a-wave amplitudes were preserved. PKCa staining of retinal cryosections from Emc3 cKO mice revealed death of rod BCs. Loss of Emc3 led to the presence of the synaptic protein mGLuR6 in the outer nuclear layer (ONL). Immunostaining analysis of presynaptic protein postsynaptic density protein 95 (PSD95) revealed rod terminals retracted to the ONL in Emc3 cKO mice at 12 months of age. In addition, deletion of Emc3 resulted in elevated glial fibrillary acidic protein, indicating reactive gliosis in the retina. Our data demonstrate that loss of Emc3 in BCs leads to decreased ERG response, increased astrogliosis and disruption of the retinal inner nuclear layer in mice of 12 months of age. Taken together, our studies indicate that Emc3 is not required for the development of BCs but is important for long-term survival of BCs.

Empathy is hypothesized to have several components, including affective, cognitive, and somatic contributors. The only validated, self-report measure to date that assesses all three forms of empathy is the Cognitive, Affective, and Somatic Empathy Scale (CASES), but no current study has reported the psychometric properties of this scale outside of the initial U.S. sample. This study reports the first psychometric analysis of a non-English translation of the CASES.Confirmatory factor analysis was used to assess the factor structure of CASES as well as its associations with callous-unemotional traits in 860 male and female children (mean age 11.54± .64 years) from the China Jintan Child Cohort Study.Analyses supported a three-factor model of cognitive, affective, and somatic empathy, with satisfactory fit indices consistent with the psychometric properties of the English version of CASES. Construct validity was established by three findings. First, females scored significantly higher in empathy than males. Second, lower scores of empathy were associated with lower IQ. Third, children with lower empathy also showed more callous-unemotional attributes.We established for the first time cross-cultural validity for Cognitive, Affective, and Somatic Empathy Scale (CASES). Our Chinese data supports the use of this new instrument in non-Western samples, and affirms the utility of this instrument for a comprehensive assessment of empathy in children.

Glaucoma is an optic neuropathy commonly associated with elevated intraocular pressure (IOP) resulting in progressive loss of retinal ganglion cells (RGCs) and optic nerve degeneration, leading to blindness. New therapeutic approaches that better preserve the visual field by promoting survival and health of RGCs are highly needed since RGC death occurs despite good IOP control in glaucoma patients. We have developed a novel approach to reliably induce chronic IOP elevation in mouse using a photopolymerizable biomatrix, hyaluronic acid glycidyl methacrylate. This is achieved by rapid in vivo crosslinking of the biomatrix at the iridocorneal angle by a flash of ultraviolet A (UVA) light to impede the aqueous outflow pathway with a controllable manner. Sustained IOP elevation was induced after a single manipulation and was maintained at ~45% above baseline for >4 weeks. Significant thinning of the inner retina and ~35% reduction in RGCs and axons was noted within one month of IOP elevation. Optic nerve degeneration showed positive correlation with cumulative IOP elevation. Activation of astrocytes and microglia appeared to be an early event in response to IOP elevation preceding detectable RGC and axon loss. Attenuated glial reactivity was noted at later stage where significant RGC/axon loss had occurred suggesting astrocytes and microglia may play different roles over the course of glaucomatous degeneration. This novel murine glaucoma model is reproducible and displays cellular changes that recapitulate several pathophysiological features of glaucoma.

This study aimed to determine the impact of stocking density on the liver proteome and cecal microbiota of Peking ducks. A total of 1,200 21-day-old ducks were randomly assigned to 5 stocking density groups of 5, 6, 7, 8 and 9 ducks/m2, with 6 replicates for each group. At 40 days of age, duck serum and pectorals were collected for biochemical tests; liver and cecal contents of ducks were gathered for proteome and microbiota analysis, respectively. Serum MDA increased while pectorals T-AOC reduced linearly with enhancing stocking density. Duck lipid metabolism was altered under different stocking density as well. Serum LDL-C increased linearly with increasing stocking density. Proteome analysis revealed fatty acid biosynthesis proteins such as acyl-CoA synthetase family member 2 and fatty acid oxidation related proteins including acyl-CoA dehydrogenase long chain and acyl-coenzyme A oxidase were enriched in high stocking density group. Additionally, high stocking density increased oxidative response associated proteins such as DDRGK domain containing 1. Furthermore, increasing stocking density diminished proteins of anti-oxidant capacity including regucalcin and catalase. 16S rDNA analysis revealed that higher stocking density was accompanied with decreased microbial diversity, as well as depletion of anti-inflammatory bacterial taxa, including Bacteroidales, Butyricimonas and Alistipe. Besides, reduced bile acid metabolism-associated bacteria such as Ruminococcaceae, Clostridiales and Desulfovibrionaceae were found in the high-density group. Both proteome and 16S rDNA results showed inflammation and chronic liver disease trend in the high-density group, which suggests the involvement of the liver-gut axis in oxidative stress.

Platelet to lymphocyte ratio (PLR) is a parameter reflecting inflammatory responses in patients with cancer. Several studies have investigated the prognostic value of PLR in patients with colorectal cancer (CRC); however, the results are controversial. Thus, we carried out a meta-analysis to evaluate the association between PLR and CRC prognostication. Relevant articles were retrieved through PubMed, Embase, and Web of Science, and pooled hazard ratio (HR) and 95% confidence interval (CI) were computed by using STATA V.12.0. Both the random-effects model and fixed-effects model were utilized. A total of 13 studies (14 cohorts) with 8,601 patients were included in the meta-analysis. Pooled HRs and 95% CIs demonstrated that increased PLR predicted poor overall survival (OS) (HR = 1.81, 95%CI:1.42-2.31, p

Turbidity (T) has been widely used to detect the occurrence of pollutants in surface water. Using data collected from January 2013 to June 2014 at eleven sites along two rivers feeding the Taihu Basin, China, the relationship between the concentration of five metals (aluminum (Al), titanium (Ti), nickel (Ni), vanadium (V), lead (Pb)) and turbidity was investigated. Metal concentration was determined using inductively coupled plasma mass spectrometry (ICP-MS). The linear regression of metal concentration and turbidity provided a good fit, with R(2) = 0.86-0.93 for 72 data sets collected in the industrial river and R(2) = 0.60-0.85 for 60 data sets collected in the cleaner river. All the regression presented good linear relationship, leading to the conclusion that the occurrence of the five metals are directly related to suspended solids, and these metal concentration could be approximated using these regression equations. Thus, the linear regression equations were applied to estimate the metal concentration using online turbidity data from January 1 to June 30 in 2014. In the prediction, the WASP 7.5.2 (Water Quality Analysis Simulation Program) model was introduced to interpret the transport and fates of total suspended solids; in addition, metal concentration downstream of the two rivers was predicted. All the relative errors between the estimated and measured metal concentration were within 30%, and those between the predicted and measured values were within 40%. The estimation and prediction process of metals' concentration indicated that exploring the relationship between metals and turbidity values might be one effective technique for efficient estimation and prediction of metal concentration to facilitate better long-term monitoring with high temporal and spatial density.

The objective of this research was to analyse whether ameliorating drought stress through exogenously applied spermine (Spm) was related to carbohydrate metabolism, dehydrins accumulation and the transcription of genes encoding dehydrins in two white clovers (drought-susceptible cv. 'Ladino' and drought-resistant cv. 'Haifa') under controlled drying conditions for 10 days. The results show that the application of Spm effectively alleviates negative effects caused by drought stress in both cultivars. Exogenous Spm led to accumulation of more water-soluble carbohydrates (WSC), sucrose, fructose and sorbitol in both cultivars under drought stress, and also significantly elevated glucose content in leaves of drought-resistant cv. 'Haifa', but had no effect on drought-susceptible cv. 'Ladino'. Accordingly, the key enzyme activities of sucrose and sorbitol metabolism changed along with the application of Spm under drought stress. Spm induced a significant increase in sucrose phosphate synthase (SPS) or sorbitol dehydrogenase (SDH) activity, but decrease in sucrose synthetase (SS) activity when two cultivars were subjected to drought. In addition, the improved accumulation of dehydrins induced by exogenous Spm coincided with three genes expression which was responsible for dehydrins synthesis. But Spm-induced transcript level of dehydrin genes increased earlier in cv. 'Ladino' than that in cv. 'Haifa'. Thus, these results suggest that ameliorating drought stress through exogenously applied Spm may be associated with increased carbohydrate accumulation and dehydrins synthesis. There are differences between drought-susceptible and -resistant white clover cultivars related to Spm regulation of WSC metabolism and dehydrins expression.

BackgroundSparganosis caused by invasion of the plerocercoid larvae (spargana) of Spirometra erinaceieuropaei have increased in recent years in China. However, the population genetic structure regarding this parasite is still unclear. In this study, we used the sequences of two mitochondrial genes cytochrome b (cytb) and cytochrome c oxidase subunit I (cox1) to analyze genetic variation and phylogeographic structure of the S. erinaceieuropaei populations.Methodology/principal findingsA total of 88 S. erinaceieuropaei isolates were collected from naturally infected frogs in 14 geographical locations of China. The complete cytb and cox1 genes of each sample was amplified and sequenced. Total 61 haplotypes were found in these 88 concatenated sequences. Each sampled population and the total population have high haplotype diversity (Hd), accompanied by very low nucleotide diversity (Pi). Phylogenetic analyses of haplotypes revealed two distinct clades (HeN+HuN+GZ-AS clade and GX+HN+GZ-GY clade) corresponding two sub-networks yielded by the median-joining network. Pairwise FST values supported great genetic differentiation between S. erinaceieuropaei populations. Both negative Fu's FS value of neutrality tests and unimodal curve of mismatch distribution analyses supported demographic population expansion in the HeN+HuN+GZ-AS clade. The BEAST analysis showed that the divergence time between the two clades took place in the early Pleistocene (1.16 Myr), and by Bayesian skyline plot (BSP) an expansion occurred after about 0.3 Myr ago.ConclusionsS. erinaceieuropaei from central and southern China has significant phylogeographic structure, and climatic oscillations during glacial periods in the Quaternary may affect the demography and diversification of this species.

BACKGROUND:Early diagnosis of human immunodeficiency virus (HIV) allows for appropriately timed interventions with improved outcomes, but HIV screening among asymptomatic persons and the general population in Singapore remains low. In 2008, Singapore's Ministry of Health implemented HIV voluntary opt-out screening (VOS) for hospitalised adults. We evaluated the outcome of VOS and surveyed reasons for its low uptake in our institution. METHODS:We assessed the outcomes of the VOS programme from January 2010 to December 2013 at National University Hospital, a 1081-bed tertiary hospital in Singapore. We also examined reasons for opting-in and opting-out using an interviewer-administered structured questionnaire in a representative sample in January 2013. RESULTS:107,523 patients fulfilled VOS criteria and were offered HIV screening, of which 5215 (4.9%) agreed to testing. 4850 (93.1%) of those who opted-in had an HIV test done. Three (0.06%) tested positive for HIV. 238 patients (14.2%) were surveyed regarding reasons for opting-in or out of VOS. 21 (8.8%) had opted-in. Patients who opted-in were likely to be younger, more educated and reported having more regular sexual partners. Type of housing, number of casual sexual partners, sexual orientation, intravenous drug use, condom use and previous sexually transmitted infection were not associated with deciding to opt-in/out. Patients' most common reasons for opting-out were: belief that they were at low risk (50.2%), belief that they were too old (26.8%), cost (6.9%) and aversion to venepuncture (6.5%). The most common reason for opting-in was desire to know their HIV status (47.6%). CONCLUSION:The success of an HIV-VOS program is largely determined by test uptake. Our study showed that the majority of eligible VOS patients opted-out of HIV screening. Given the considerable cost and low yield of this programme, more needs to be done to better equip patients in self-risk assessment and opting in to testing.

Take-all, which is caused by the fungal pathogen, Gaeumannomyces graminis var. tritici (Ggt), is an important soil-borne root rot disease of wheat occurring worldwide. However, the genetic basis of Ggt pathogenicity remains unclear. In this study, transcriptome sequencing for Ggt in axenic culture and Ggt-infected wheat roots was performed using Illumina paired-end sequencing. Approximately 2.62 and 7.76 Gb of clean reads were obtained, and 87% and 63% of the total reads were mapped to the Ggt genome for RNA extracted from Ggt in culture and infected roots, respectively. A total of 3,258 differentially expressed genes (DEGs) were identified with 2,107 (65%) being 2-fold up-regulated and 1,151 (35%) being 2-fold down-regulated between Ggt in culture and Ggt in infected wheat roots. Annotation of these DEGs revealed that many were associated with possible Ggt pathogenicity factors, such as genes for guanine nucleotide-binding protein alpha-2 subunit, cellulase, pectinase, xylanase, glucosidase, aspartic protease and gentisate 1, 2-dioxygenase. Twelve DEGs were analyzed for expression by qRT-PCR, and could be generally divided into those with high expression only early in infection, only late in infection and those that gradually increasing expression over time as root rot developed. This indicates that these possible pathogenicity factors may play roles during different stages of the interaction, such as signaling, plant cell wall degradation and responses to plant defense compounds. This is the first study to compare the transcriptomes of Ggt growing saprophytically in axenic cultures to it growing parasitically in infected wheat roots. As a result, new candidate pathogenicity factors have been identified, which can be further examined by gene knock-outs and other methods to assess their true role in the ability of Ggt to infect roots.

Compounds released by mungbean roots potentially represent an enormous source of nitrogen (N) and carbon (C) in mungbean-oat intercropping systems. In this study, an in situ experiment was conducted using a 15N - 13C double stem-feeding method to measure N and C derived from the rhizodeposition (NdfR and CdfR) of mungbean and their transfer to oats in an intercropping system. Mungbean plants were sole cropped (S) or intercropped (I) with oat. The plants were labeled 5 weeks after planting and were harvested at the beginning of pod setting (Ip and Sp) and at maturity (Im and Sm). More than 60% and 50% of the applied 15N and 13C, respectively, were recovered in each treatment, with 15N and 13C being quite uniformly distributed in the different plant parts. NdfR represented 9.8% (Sp), 9.2% (Ip), 20.1% (Sm), and 21.2% (Im) of total mungbean plant N, whereas CdfR represented 13.3% (Sp), 42.0% (Ip), 15.4% (Sm), and 22.6% (Im) of total mungbean plant C. When considering the part of rhizodeposition transferred to associated oat, intercropping mungbean released more NdfR and CdfR than mungbean alone. About 53.4-83.2% of below-ground plant N (BGP-N) and 58.4-85.9% of BGP-C originated from NdfR and CdfR, respectively. The N in oats derived from mungbean increased from 7.6% at the pod setting stage to 9.7% at maturity, whereas the C in oats increased from 16.2% to 22.0%, respectively. Only a small percentage of rhizodeposition from mungbean was transferred to oats in the intercropping systems, with a large percentage remaining in the soil. This result indicates that mungbean rhizodeposition might contribute to higher N and C availability in the soil for subsequent crops.

A traditional traffic signal control system is established based on vehicular delay, queue length, saturation and other indicators. However, due to the increasing severity of urban environmental pollution issues and the development of a resource-saving and environmentally friendly social philosophy, the development of low-carbon and energy-efficient urban transport is required. This paper first defines vehicular trajectories and the calculation of vehicular emissions based on VSP. Next, a regression analysis method is used to quantify the relationship between vehicular emissions and delay, and a traffic signal control model is established to reduce emissions and delay using the enumeration method combined with saturation constraints. Finally, one typical intersection of Changchun is selected to verify the model proposed in this paper; its performance efficiency is also compared using simulations in VISSIM. The results of this study show that the proposed model can significantly reduce vehicle delay and traffic emissions simultaneously.

Microtubule-associated proteins play a crucial role in the regulation of microtubule dynamics, and are very important for plant cell and organ development. SBgLR is a potato pollen-specific protein, with five imperfect V-V-E-K-K-N/E-E repetitive motifs that are responsible for microtubule binding activity. In present study, SBgLR showed typical microtubule-associated protein characteristics; it bound tubulin and microtubules, and colocalized with microtubules in vitro. We also found that SBgLR could form oligomers, and that both the SBgLR monomers and oligomers bundle microtubules in vitro. Constitutive expression of SBgLR in tobacco caused curving and right-handed twisting root growth, abnormal directional cell expansion and cell layer arrangement, and pollen abortion. Immunofluorescence staining assays revealed that microtubule organization is altered in root epidermal cells in SBgLR-overexpressing lines. These suggest that SBgLR functions as a microtubule-associated protein in pollen development. Our results indicate that normal organization of MTs may be crucial for pollen development.

PURPOSE: The causal genes for congenital cataract are good candidates for the genetic susceptibility for age-related cataract (ARC). The aim of this study was to investigate association between the polymorphisms in the causal genes for congenital cataract and ARC in a Chinese population. Meanwhile, we performed the replication study for previous identified risk genes for ARC. METHODS: We recruited 212 sporadic Han Chinese patients with age-related cataracts (ARC) and 172 normal controls in this study. We analyzed 31 SNPs from 13 genes which mostly possible contributes the progress of ARC in a Chinese population, comprising 212 cataract patients and 172 controls. Polymorphism-spanning fragments were amplified by using the multiplex polymerase chain reaction (PCR) and genotyped using primer extension method in MassARRAY platform. Allelic and haplotypic difference in the frequencies were estimated using the SHEsis software platform. P-value was adjusted by the Bonferroni correction. RESULTS: There was no difference in the frequencies of the genotype and allele of the all SNPs between the patients with ARC and the controls. In the haplotypic analysis, the haplotypes consisting of rs7154572, rs7150141 and rs12432994 in Kinesin Light Chain 1 Gene (KLC1) showed significant association with ARC (p = 0.000878). A rare haplotype CGT was more frequent in patients (p = 0.000106, and p = 0.00795 after corrected for 75 tests). CONCLUSIONS: Our study provides evidence that the combined effect of three variants within the KLC1 gene may predispose to ARC, but the precise mechanism needs further investigating.

Prodigiosins (PGs) are a family of natural red pigments with anticancer activity, and one member of the family has entered clinical phase II trials. However, the anticancer mechanisms of PGs remain largely unclear. This study was designed to investigate the molecular basis of anticancer activity of UP, a derivative of PGs, in P388 cells. By introducing pharmacological inhibitors and utilizing a variety of analytical approaches including western blotting, flow cytometry and confocal laser microscopy, we found that UP inhibited proliferation of P388 via arresting cells at G2/M phase and inducing cells apoptosis, which was related to the activation of P38, JNK rather than ERK1/2 signaling. ROS regeneration and acidification in cells appear not involved in UP induced apoptosis. Furthermore, utilizing mass spectrometry, sucrose density gradient fractionation and immunofluorescence staining, we discovered that UP was apparently located at ribosome. These results together indicate that ribosome may be the potential target of UP in cancer cells, which opened a new avenue in delineating the anticancer mechanism of PGs.

Bone morphogenetic protein (BMP) signaling pathway plays important roles in urinary tract development although the detailed regulation of its activity in this process remains unclear. Here we report that follistatin-like 1 (Fstl1), encoding a secreted extracellular glycoprotein, is expressed in developing ureter and antagonizes BMP signaling activity. Mouse embryos carrying disrupted Fstl1 gene displayed prominent hydroureter arising from proximal segment and ureterovesical junction defects. These defects were associated with significant reduction in ureteric epithelial cell proliferation at E15.5 and E16.5 as well as absence of subepithelial ureteral mesenchymal cells in the urinary tract at E16.5 and E18.5. At the molecular level, increased BMP signaling was found in Fstl1 deficient ureters, indicated by elevated pSmad1/5/8 activity. In vitro study also indicated that Fstl1 can directly bind to ALK6 which is specifically expressed in ureteric epithelial cells in developing ureter. Furthermore, Sonic hedgehog (SHH) signaling, which is crucial for differentiation of ureteral subepithelial cell proliferation, was also impaired in Fstl1(-/-) ureter. Altogether, our data suggest that Fstl1 is essential in maintaining normal ureter development by antagonizing BMP signaling.

A sudden mechanical insult to the spinal cord is usually caused by changing pressure on the surface of the spinal cord. Most of these insults are mechanical force injuries, and their mechanism of injury to the spinal cord is largely unknown.Using a compression-driven instrument to simulate mechanical force, we applied mechanical pressure of 0.5 MPa to rat dorsal root ganglion (DRG) neurons for 10 min to investigate cytoskeletal alterations and calpain-induced apoptosis after the mechanical force injury.The results indicated that mechanical forces affect the structure of the cytoskeleton and cell viability, induce early apoptosis, and affect the cell cycle of DRG neurons. In addition, the calpain inhibitor PD150606 reduced cytoskeletal degradation and the rate of apoptosis after mechanical force injury.Thus, calpain may play an important role in DRG neurons in the regulation of apoptosis and cytoskeletal alterations induced by mechanical force. Moreover, cytoskeletal alterations may be substantially involved in the mechanotransduction process in DRG neurons after mechanical injury and may be induced by activated calpain. To our knowledge, this is the first report to demonstrate a relationship between cytoskeletal degradation and apoptosis in DRG neurons.

BackgroundEnvironmental stress can accelerate the directional selection and evolutionary rate of specific stress-response proteins to bring about new or altered functions, enhancing an organism's fitness to challenging environments. Plateau pika (Ochotona curzoniae), an endemic and keystone species on Qinghai-Tibetan Plateau, is a high hypoxia and low temperature tolerant mammal with high resting metabolic rate and non-shivering thermogenesis to cope in this harsh plateau environment. Leptin is a key hormone related to how these animals regulate energy homeostasis. Previous molecular evolutionary analysis helped to generate the hypothesis that adaptive evolution of plateau pika leptin may be driven by cold stress.Methodology/principal findingsTo test the hypothesis, recombinant pika leptin was first purified. The thermogenic characteristics of C57BL/6J mice injected with pika leptin under warm (23±1°C) and cold (5±1°C) acclimation is investigated. Expression levels of genes regulating adaptive thermogenesis in brown adipose tissue and the hypothalamus are compared between pika leptin and human leptin treatment, suggesting that pika leptin has adaptively and functionally evolved. Our results show that pika leptin regulates energy homeostasis via reduced food intake and increased energy expenditure under both warm and cold conditions. Compared with human leptin, pika leptin demonstrates a superior induced capacity for adaptive thermogenesis, which is reflected in a more enhanced β-oxidation, mitochondrial biogenesis and heat production. Moreover, leptin treatment combined with cold stimulation has a significant synergistic effect on adaptive thermogenesis, more so than is observed with a single cold exposure or single leptin treatment.Conclusions/significanceThese findings support the hypothesis that cold stress has driven the functional evolution of plateau pika leptin as an ecological adaptation to the Qinghai-Tibetan Plateau.

The long-term fate of stem cells after intramyocardial delivery is unknown. We used noninvasive, repetitive PET/CT imaging with [(18)F]FEAU to monitor the long-term (up to 5 months) spatial-temporal dynamics of MSCs retrovirally transduced with the sr39HSV1-tk gene (sr39HSV1-tk-MSC) and implanted intramyocardially in pigs with induced acute myocardial infarction. Repetitive [(18)F]FEAU PET/CT revealed a biphasic pattern of sr39HSV1-tk-MSC dynamics; cell proliferation peaked at 33-35 days after injection, in periinfarct regions and the major cardiac lymphatic vessels and lymph nodes. The sr39HSV1-tk-MSC-associated [(18)F]FEAU signals gradually decreased thereafter. Cardiac lymphography studies using PG-Gd-NIRF813 contrast for MRI and near-infrared fluorescence imaging showed rapid clearance of the contrast from the site of intramyocardial injection through the subepicardial lymphatic network into the lymphatic vessels and periaortic lymph nodes. Immunohistochemical analysis of cardiac tissue obtained at 35 and 150 days demonstrated several types of sr39HSV1-tk expressing cells, including fibro-myoblasts, lymphovascular cells, and microvascular and arterial endothelium. In summary, this study demonstrated the feasibility and sensitivity of [(18)F]FEAU PET/CT imaging for long-term, in-vivo monitoring (up to 5 months) of the fate of intramyocardially injected sr39HSV1-tk-MSC cells. Intramyocardially transplanted MSCs appear to integrate into the lymphatic endothelium and may help improve myocardial lymphatic system function after MI.

MHC class I molecules present a comprehensive mixture of peptides on the cell surface for immune surveillance. The peptides represent the intracellular protein milieu produced by translation of endogenous mRNAs. Unexpectedly, the peptides are encoded not only in conventional AUG initiated translational reading frames but also in alternative cryptic reading frames. Here, we analyzed how ribosomes recognize and use cryptic initiation codons in the mRNA. We find that translation initiation complexes assemble at non-AUG codons but differ from canonical AUG initiation in response to specific inhibitors acting within the peptidyl transferase and decoding centers of the ribosome. Thus, cryptic translation at non-AUG start codons can utilize a distinct initiation mechanism which could be differentially regulated to provide peptides for immune surveillance.

BACKGROUND: Environmental stress can accelerate the evolutionary rate of specific stress-response proteins and create new functions specialized for different environments, enhancing an organism's fitness to stressful environments. Pikas (order Lagomorpha), endemic, non-hibernating mammals in the modern Holarctic Region, live in cold regions at either high altitudes or high latitudes and have a maximum distribution of species diversification confined to the Qinghai-Tibet Plateau. Variations in energy metabolism are remarkable for them living in cold environments. Leptin, an adipocyte-derived hormone, plays important roles in energy homeostasis. METHODOLOGY/PRINCIPAL FINDINGS: To examine the extent of leptin variations within the Ochotona family, we cloned the entire coding sequence of pika leptin from 6 species in two regions (Qinghai-Tibet Plateau and Inner Mongolia steppe in China) and the leptin sequences of plateau pikas (O. curzonia) from different altitudes on Qinghai-Tibet Plateau. We carried out both DNA and amino acid sequence analyses in molecular evolution and compared modeled spatial structures. Our results show that positive selection (PS) acts on pika leptin, while nine PS sites located within the functionally significant segment 85-119 of leptin and one unique motif appeared only in pika lineages-the ATP synthase alpha and beta subunit signature site. To reveal the environmental factors affecting sequence evolution of pika leptin, relative rate test was performed in pikas from different altitudes. Stepwise multiple regression shows that temperature is significantly and negatively correlated with the rates of non-synonymous substitution (Ka) and amino acid substitution (Aa), whereas altitude does not significantly affect synonymous substitution (Ks), Ka and Aa. CONCLUSIONS/SIGNIFICANCE: Our findings support the viewpoint that adaptive evolution may occur in pika leptin, which may play important roles in pikas' ecological adaptation to extreme environmental stress. We speculate that cold, and probably not hypoxia, may be the primary environmental factor for driving adaptive evolution of pika leptin.

Big data pertains to extensive and intricate compilations of information that necessitate the implementation of proficient and cost-effective evaluation and analysis tools to derive insights and support decision making. The Fermatean fuzzy set theory possesses remarkable capability in capturing imprecision due to its capacity to accommodate complex and ambiguous problem descriptions. This paper presents the study of the concepts of dynamic ordered weighted aggregation operators in the context of Fermatean fuzzy environment. In numerous practical decision making scenarios, the term "dynamic" frequently denotes the capability of obtaining decision-relevant data at various time intervals. In this study, we introduce two novel aggregation operators: Fermatean fuzzy dynamic ordered weighted averaging and geometric operators. We investigate the attributes of these operators in detail, offering a comprehensive description of their salient features. We present a step-by-step mathematical algorithm for decision making scenarios in the context of proposed methodologies. In addition, we highlight the significance of these approaches by presenting the solution to the decision making problem and determining the most effective big data analytics platform for YouTube data analysis. Finally, we perform a thorough comparative analysis to assess the effectiveness of the suggested approaches in comparison to a variety of existing techniques., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Alghazzawi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Road traffic accidents (RTAs) pose a significant hazard to the security of the general public, especially in developing nations. A daily average of more than three thousand fatalities is recorded worldwide, rating it as the second most prevalent cause of death among people aged 5-29. Precise and reliable decisionmaking techniques are essential for identifying the most effective approach to mitigate road traffic incidents. This research endeavors to investigate this specific concern. The Fermatean fuzzy set (FFS) is a strong and efficient method for addressing ambiguity, particularly when the concept of Pythagorean fuzzy set fails to provide a solution. This research presents two innovative aggregation operators: the Fermatean fuzzy ordered weighted averaging (FFOWA) operator and the Fermatean fuzzy dynamic ordered weighted geometric (FFOWG) operator. The salient characteristics of these operators are discussed and important exceptional scenarios are thoroughly delineated. Furthermore, by implementing the suggested operators, we develop a systematic approach to handle multiple attribute decisionmaking (MADM) scenarios that involve Fermatean fuzzy (FF) data. In order to show the viability of the developed method, we provide a numerical illustration encompassing the determination of the most effective approach to alleviate road traffic accidents. Lastly, we conduct a comparative evaluation of the proposed approach in relation to a number of established methodologies., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Alghazzawi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Alveolar echinococcosis (AE) is a lethal parasitic disease. In Gansu Province of China, all AE cases reported in literature were from Zhang and Min Counties, the southern part of the province. Here, we report the discovery of nine AE cases and one cystic echinococcosis (CE) case from Nanfeng Town of Minle County, in the middle of Hexi Corridor in west Gansu Province. The diagnosis of these cases were confirmed by serology, histopathology, computed tomography, B-ultrasound, immunohistochemistry method, DNA polymerase chain reaction and sequencing analysis. Because eight of nine AE cases came from First Zhanglianzhuang (FZLZ) village, we conducted preliminary epidemiological analyses of 730 persons on domestic water, community and ecology such as 356 dogs' faeces of FZLZ, in comparison with those of other five villages surrounding FZLZ. Our studies indicate that Nanfeng Town of Minle County is a newly discovered focus of AE in China as a CE and AE co-epidemic area. Further research of Echinococcus multilocularis transmission pattern in the area should be carried for prevention of this parasitic disease.

Objectives: Clinical risk stratification has an important function in preoperative evaluation of patients at risk for cardiac events prior to non-cardiac surgery. The aim of this study was to determine whether the combined measurement of pre-operative N-terminal pro-brain natriuretic peptide (NT-pro-BNP) and cardiac troponin I (cTnI) could provide useful prognostic information about postoperative major adverse cardiac events (MACE) within 30 days in patients aged over 60 years undergoing emergent non-cardiac surgery., Methods: The study group comprised 2519 patients aged over 60 years that were undergoing emergent non-cardiac surgery between December 2007 and December 2013. NT-pro-BNP and cTnI were measured during hospital admission. The patients were monitored for MACE (cardiac death, non-fatal myocardial infarction, or cardiac arrest) during the 30-day postoperative follow-up period., Results: MACE occurred in 251 patients (10.0%). Preoperative NT-pro-BNP and cTNI level were significantly higher in the individuals that experienced MACE than in those who did not (P < 0.001). The confounding factors of age, sex, co-morbidities and preoperative medications were adjusted in a multivariate logistic regression analysis. This analysis showed that preoperative NT-proBNP level > 917 pg/mL (OR 4.81, 95% CI 3.446-6.722, P < 0.001) and cTnI ≥ 0.07 ng/mL (OR 8.74, 95% CI 5.881-12.987, P < 0.001) remained significantly and independently associated with MACE after the adjustment of the confounding factors. Kaplan-Meier event-free survival curves demonstrated that patients with preoperative simultaneous NT-proBNP level > 917 pg/mL and cTnT ≥ 0.07 ng/mL had worse event-free survival than individual assessments of either biomarker., Conclusion: Preoperative plasma NT-proBNP and cTnI are both independently associated with an increased risk of MACE in elderly patients after emergent non-cardiac surgery. The combination of these biomarkers provides better prognostic information than using either biomarker separately.

To search for more effective tuberculosis (TB) subunit vaccines, antigens expressed in different growth stages of Mycobacterium tuberculosis (M. tuberculosis), such as RpfE (Rv2450c) produced in the stage of resuscitation, Mtb10.4 (Rv0288), Mtb8.4 (Rv1174c), ESAT6 (Rv3875), Ag85B (Rv1886c) mainly secreted by replicating bacilli, and HspX (Rv2031c) highly expressed in dormant bacilli, were selected to construct six fusion proteins: ESAT6-Ag85B-MPT64190-198-Mtb8.4 (EAMM), Mtb10.4-HspX (MH), ESAT6-Mtb8.4, Mtb10.4-Ag85B, ESAT6-Ag85B, and ESAT6-RpfE. The six fusion proteins were separately emulsified in an adjuvant composed of N,N'-dimethyl-N, N'-dioctadecylammonium bromide (DDA), polyribocytidylic acid (poly I:C) and gelatin to construct subunit vaccines, and their protective effects against M. tuberculosis infection were evaluated in C57BL/6 mice. Furthermore, the boosting effects of EAMM and MH in the adjuvant of DDA plus trehalose 6,6'-dimycolate (TDM) on BCG-induced immunity were also evaluated. It was found that the six proteins were stably produced in E. coli and successfully purified by chromatography. Among them, EAMM presented the most effective protection against M. tuberculosis. Interestingly, the mice that received EAMM+MH had significantly lower bacterial counts in the lungs and spleens than the single protein vaccinated groups, and had the same effect as those that received BCG. In addition, EAMM and MH could improve BCG-primed protective efficacy against M. tuberculosis infection in mice. In conclusion, the combination of EAMM and MH containing antigens from both replicating and dormant stages of the bacilli could induce robust immunity against M. tuberculosis infection in mice and may serve as promising subunit vaccine candidate.

The phytohormone abscisic acid ((+)-ABA) plays a key role in many processes. The biological and biochemical activities of unnatural (-)-ABA have been extensively investigated since 1960s. However, the recognition mechanism by which only a few members among PYR/PYL/RCAR (PYLs) family can bind (-)-ABA remains largely unknown. Here we systematically characterized the affinity of PYLs binding to the (-)-ABA and reported the crystal structures of apo-PYL5, PYL3-(-)-ABA and PYL9-(+)-ABA. PYL5 showed the strongest binding affinity with (-)-ABA among all the PYLs. PYL9 is a stringently exclusive (+)-ABA receptor with interchangeable disulfide bonds shared by a subclass of PYLs. PYL3 is a dual receptor to both ABA enantiomers. The binding orientation and pocket of (-)-ABA in PYLs are obviously different from those of (+)-ABA. Steric hindrance and hydrophobic interaction are the two key factors in determining the stereospecificity of PYLs binding to (-)-ABA, which is further confirmed by gain-of-function and loss-of-function mutagenesis. Our results provide novel insights of the bioactivity of ABA enantiomers onto PYLs, and shed light on designing the selective ABA receptors agonists.

Background: Amino acid sequence alignment of phage phiC31 integrase with the serine recombinases family revealed highly conserved regions outside the catalytic domain. Until now, no system mutational or biochemical studies have been carried out to assess the roles of these conserved residues in the recombination of phiC31 integrase., Methodology/principal Findings: To determine the functional roles of these conserved residues, a series of conserved residues were targeted by site-directed mutagenesis. Out of the 17 mutants, 11 mutants showed impaired or no recombination ability, as analyzed by recombination assay both in vivo and in vitro. Results of DNA binding activity assays showed that mutants (R18A, I141A, L143A,E153A, I432A and V571A) exhibited a great decrease in DNA binding affinity, and mutants (G182A/F183A, C374A, C376A/G377A, Y393A and V566A) had completely lost their ability to bind to the specific target DNA attB as compared with wild-type protein. Further analysis of mutants (R18A, I141A, L143A and E153A) synapse and cleavage showed that these mutants were blocked in recombination at the stage of strand cleavage., Conclusions/significance: This data reveals that some of the highly conserved residues both in the N-terminus and C-terminus region of phiC31 integrase, play vital roles in the substrate binding and cleavage. The cysteine-rich motif and the C-tail val-rich region of phiC31 integrase may represent the major DNA binding domains of phiC31 integrase.