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Honey bees are a model for host-microbial interactions with experimental designs evolving towards conventionalized worker bees. Research on gut microbiome transmission and assembly has examined only a fraction of factors associated with the colony and hive environment. Here we studied the effects of diet and social isolation on tissue-specific bacterial and fungal colonization of the midgut and two key hindgut regions. We found that both treatment factors significantly influenced early hindgut colonization explaining similar proportions of microbiome variation. In agreement with previous work, social interaction with older workers was unnecessary for core hindgut bacterial transmission. Exposure to natural eclosion and fresh stored pollen resulted in gut bacterial communities that were taxonomically and structurally equivalent to those produced in the natural colony setting. Stressed diets of no pollen or autoclaved pollen in social isolation resulted in decreased fungal abundance and bacterial diversity, and atypical microbiome structure and tissue-specific variation of functionally important core bacteria. Without exposure to the active hive environment, the abundance and strain diversity of keystone ileum species Gilliamella apicola was markedly reduced. These changes were associated with significantly larger ileum microbiotas suggesting that extended exposure to the active hive environment plays an antibiotic role in hindgut microbiome establishment. We conclude that core hindgut microbiome transmission is facultative horizontal with 5 of 6 core hindgut species readily acquired from the built hive structure and natural diet. Our findings contribute novel insights into factors influencing assembly and maintenance of honey bee gut microbiota and facilitate future experimental designs.

As essential pollinators of ecosystems and agriculture, honey bees (Apis mellifera) are host to a variety of pathogens that result in colony loss. Two highly prevalent larval diseases are European foulbrood (EFB) attributed to the bacterium Melissococcus plutonius, and Varroosis wherein larvae can be afflicted by one or more paralytic viruses. Here we used high-throughput sequencing and qPCR to detail microbial succession of larval development from six diseased, and one disease-free apiary. The disease-free larval microbiome revealed a variety of disease-associated bacteria in early larval instars, but later developmental stages were dominated by beneficial symbionts. Microbial succession associated with EFB pathology differed by apiary, characterized by associations with various gram-positive bacteria. At one apiary, diseased larvae were uniquely described as “melting and deflated”, symptoms associated with Varroosis. We found that Acute Bee Paralysis Virus (ABPV) levels were significantly associated with these symptoms, and various gram-negative bacteria became opportunistic in the guts of ABPV afflicted larvae. Perhaps contributing to disease progression, the ABPV associated microbiome was significantly depleted of gram-positive bacteria, a likely result of recent antibiotic application. Our results contribute to the understanding of brood disease diagnosis and treatment, a growing problem for beekeeping and agriculture worldwide.

Supplemental material, sj-docx-1-jdr-10.1177_00220345231164104 for FoxO1/NLRP3 Inflammasome Promotes Age-Related Alveolar Bone Resorption by Z. Wang, F. Zhou, X. Feng, H. Li, C. Duan, Y. Wu and Y. Xiong in Journal of Dental Research

Honey bees exhibit an elaborate social structure based in part on an age-related division of labor. Young workers perform tasks inside the hive, while older workers forage outside the hive, tasks associated with distinct diets and metabolism. Critical to colony fitness, the work force can respond rapidly to changes in the environment or colony demography and assume emergency tasks, resulting in young foragers or old nurses. We hypothesized that both task and age affect the gut microbiota consistent with changes to host diet and physiology. We performed two experiments inducing precocious foragers and reverted nurses, then quantified tissue-specific gut microbiota and host metabolic state associated with nutrition, immunity and oxidative stress. In the precocious forager experiment, both age and ontogeny explained differences in midgut and ileum microbiota, but host gene expression was best explained by an interaction of these factors. Precocious foragers were nutritionally deficient, and incurred higher levels of oxidative damage relative to age-matched nurses. In the oldest workers, reverted nurses, the oxidative damage associated with age and past foraging was compensated by high Vitellogenin expression, which exceeded that of young nurses. Host-microbial interactions were evident throughout the dataset, highlighted by an age-based increase of Gilliamella abundance and diversity concurrent with increased carbonyl accumulation and CuZnSOD expression. The results in general contribute to an understanding of ecological succession of the worker gut microbiota, defining the species-level transition from nurse to forager.

Sirui Fu,1,2,&ast; Meiqing Pan,2,3,&ast; Jie Zhang,4,&ast; Hui Zhang,2,3,5,&ast; Zhenchao Tang,2,3,5 Yong Li,1 Wei Mu,2,3,5 Jianwen Huang,1 Di Dong,3 Chongyang Duan,6 Xiaoqun Li,7 Shuo Wang,2,3 Xudong Chen,8 Xiaofeng He,9 Jianfeng Yan,10 Ligong Lu,1 Jie Tian2,3,5,11 1Zhuhai Interventional Medical Centre, Zhuhai People’s Hospital (Zhuhai Hospital Affiliated with Jinan University), Zhuhai, People’s Republic of China; 2Beijing Advanced Innovation Centre for Big Data-Based Precision Medicine, School of Engineering Medicine, Beihang University, Beijing, People’s Republic of China; 3CAS Key Laboratory of Molecular Imaging, Beijing Key Laboratory of Molecular Imaging, The State Key Laboratory of Management and Control for Complex Systems, Institute of Automation, Chinese Academy of Sciences, Beijing, People’s Republic of China; 4Department of Radiology, Zhuhai People’s Hospital (Zhuhai Hospital Affiliated with Jinan University), Zhuhai, People’s Republic of China; 5Key Laboratory of Big Data-Based Precision Medicine (Beihang University), Ministry of Industry and Information Technology, Beijing, People’s Republic of China; 6Department of Biostatistics, School of Public Health, Southern Medical University, Guangzhou, People’s Republic of China; 7Department of Interventional Treatment, Zhongshan City People’s Hospital, Zhongshan, People’s Republic of China; 8Department of Radiology, Shenzhen People’s Hospital, Shenzhen, People’s Republic of China; 9Interventional Diagnosis and Treatment Department, Nanfang Hospital, Southern Medical University, Guangzhou, People’s Republic of China; 10Department of Radiology, Yangjiang People’s Hospital, Yangjiang, People’s Republic of China; 11University of Chinese Academy of Sciences, Beijing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jie TianBeijing Advanced Innovation Centre for Big Data-Based Precision Medicine, School of Engineering Medicine, Beihang University, Beijing, 100191, People’s Republic of ChinaEmail tian@ieee.orgLigong LuZhuhai Interventional Medical Centre, Zhuhai People’s Hospital (Zhuhai Hospital Affiliated with Jinan University), No. 79 Kangning Road, Zhuhai, 519000, Guangdong Province, People’s Republic of ChinaEmail llg0902@sina.comPurpose: For timely treatment of extrahepatic metastasis and macrovascular invasion (aggressive progressive disease [PD]) in hepatocellular carcinoma, models aimed at stratifying the risks of subsequent aggressive PD should be constructed.Patients and Methods: After dividing 332 patients from five hospitals into training (n = 236) and validation (n = 96) datasets, non-invasive models, including clinical/semantic factors (ModelCS), deep learning radiomics (ModelD), and both (ModelCSD), were constructed to stratify patients according to the risk of aggressive PD. We examined the discrimination and calibration; similarly, we plotted a decision curve and devised a nomogram. Furthermore, we performed analyses of subgroups who received different treatments or those in different disease stages and compared time to aggressive PD and overall survival in the high- and low-risk subgroups.Results: Among the constructed models, ModelCSD, combining clinical/semantic factors and deep learning radiomics, outperformed ModelCS and ModelD (areas under the curve [AUCs] for the training dataset: 0.741, 0.815, and 0.856; validation dataset: 0.780, 0.836, and 0.862), with statistical difference per the net reclassification improvement, the integrated discrimination improvement, and/or the DeLong test in both datasets. Besides, ModelCSD had the best calibration and decision curves. The performance of ModelCSD was not affected by treatment types (AUC: resection = 0.839; transarterial chemoembolization = 0.895; p = 0.183) or disease stages (AUC: BCLC [Barcelona Clinic Liver Cancer] stage 0 and A = 0.827; BCLC stage AB &B = 0.861; p = 0.537). Moreover, the high-risk group had a significantly shorter median time to aggressive PD than the low-risk group (training dataset hazard ratio [HR] = 0.108, p < 0.001; validation dataset HR = 0.058, p < 0.001) and poorer overall survival (training dataset HR = 0.357, p < 0.001; validation dataset HR = 0.204, p < 0.001).Conclusion: Our deep learning-based model successfully stratified the risks of aggressive PD. In the high-risk population, current guideline indicates that first-line treatments are insufficient to prevent extrahepatic metastasis and macrovascular invasion and ensure survival benefits, so more therapies may be explored for these patients.Keywords: aggressive disease progression, deep learning radiomics, clinical factors, high-risk, risk prediction

Fa Jin, Weiyang Ou, Boyang Wei, Haiyan Fan, Chengcong Wei, Dazhao Fang, Guangxu Li, Wenchao Liu, Jiahui Liu, Lei Jin, Xuying He, Chuanzhi Duan Neurosurgery Center, Department of Cerebrovascular Surgery, The National Key Clinical Specialty, The Engineering Technology Research Center of Education Ministry of China on Diagnosis and Treatment of Cerebrovascular Disease, Guangdong Provincial Key Laboratory on Brain Function Repair and Regeneration, The Neurosurgery Institute of Guangdong Province, Zhujiang Hospital, Southern Medical University, Guangzhou, 510282, People’s Republic of ChinaCorrespondence: Chuanzhi Duan; Xuying He Tel +86 135 399 622 33; +86 136 888 771 33Fax +86 206 164 3010Email doctor_duanzj@163.com; 2517079319@qq.comBackground: Ischemic stroke is one of the leading causes of mortality and disability worldwide. Following stroke, there is secondary neuroinflammation that promotes further injury. Identifying the long non-coding RNA (lncRNA) involved in neuroinflammation after cerebral ischemic stroke will promote the discovery of potential therapeutic targets.Methods: We identified differentially expressed genes from genome-wide RNA-seq profiles of mice with focal ischemia using Gene Ontology Term Enrichment, Kyoto Encyclopedia of Genes and Genomes, and Gene Set Enrichment analyses. Immune cell infiltration deconvolution, protein-protein interaction network construction, and co-expression network analyses were also used to screen lncRNAs. In further experiments, lncRNA Neat1 knockdown animal models were developed by intraventricular injection of the antisense oligonucleotide before performing middle cerebral artery occlusion (MCAO). An enzyme-linked immunosorbent assay was performed to measure the level of cytokines. Hematoxylin-eosin staining and immunohistochemical staining were used to observe the changes in morphology.Results: Enrichment analysis revealed that differential mRNAs induced neuroinflammation after MCAO. Immune deconvolution showed that the proportion of microglia gradually increased while monocytes decreased within 24 h. We identified six hub lncRNAs (Neat1, Gm10827, Trp53cor1, Mir670hg, C730002L08Rik, and Mir181a-hg) that were highly correlated with activated-microglia mRNAs (cor > 0.8). We found that Neat1 had the highest correlation coefficient with pro-inflammatory factor mRNA levels. In vivo experiments demonstrated that Neat1 had abnormally high expression after MCAO. Knockdown of Neat1 could significantly alleviate brain damage by reducing the number of activated microglia and reducing their release of proinflammatory cytokines.Conclusion: We identified inflammation-associated lncRNA Neat1 as crucial, which means it is a potential target for ischemic stroke treatment.Keywords: bioinformatics, lncRNA Neat1, ischemic stroke, MCAO, microglia, neuroinflammation

Honey bees exhibit an elaborate social structure based in part on an age-related division of labor. Young workers perform tasks inside the hive, while older workers forage outside the hive, tasks associated with distinct diets and metabolism. Critical to colony fitness, the work force can respond rapidly to changes in the environment or colony demography and assume emergency tasks, resulting in young foragers or old nurses. We hypothesized that both task and age affect the gut microbiota consistent with changes to host diet and physiology. We performed two experiments inducing precocious foragers and reverted nurses, then quantified tissue-specific gut microbiota and host metabolic state associated with nutrition, immunity and oxidative stress. In the precocious forager experiment, both age and ontogeny explained differences in midgut and ileum microbiota, but host gene expression was best explained by an interaction of these factors. Precocious foragers were nutritionally deficient, and incurred higher levels of oxidative damage relative to age-matched nurses. In the oldest workers, reverted nurses, the oxidative damage associated with age and past foraging was compensated by high Vitellogenin expression, which exceeded that of young nurses. Host-microbial interactions were evident throughout the dataset, highlighted by an age-based increase of

The highly social honey bee has dense populations but a significantly reduced repertoire of immune genes relative to solitary species, suggesting a greater reliance on social immunity. Here we investigate immune gene expression and gut microbial succession in queens during colony introduction. Recently mated queens were placed into an active colony or a storage hive for multiple queens: a queen-bank. Feeding intensity, social context, and metabolic demand differ greatly between the two environments. After 3 weeks, we examined gene expression associated with oxidative stress and immunity and performed high-throughput sequencing of the queen gut microbiome across four alimentary tract niches. Microbiota and gene expression in the queen hindgut differed by time, queen breeder source, and metabolic environment. In the ileum, upregulation of most immune and oxidative stress genes occurred regardless of treatment conditions, suggesting postmating effects on gut gene expression. Counterintuitively, queens exposed to the more social colony environment contained significantly less bacterial diversity indicative of social immune factors shaping the queens microbiome. Queen bank queens resembled much older queens with decreased Alpha 2.1, greater abundance of

Rhabdomyosarcoma (RMS) is one of the most common soft tissue sarcomas and is extremely aggressive and highly malignant. The majority of cases occur in children, and most commonly appear in the arms or legs. Currently, RMS is treated with aggressive chemotherapy, as well as surgery and/or radiation in many cases. The aggressive treatments often cause long term aftereffects, and can even cause permanent life-altering disabilities. Alternative treatment modalities are urgently needed. The aim of this work is to build a dual-functional therapy/near-infrared (NIR) imaging nanoparticle, which could perform gene therapy with specific RMS gene knock-down regulated by pH-activated-release, and monitor the effects on tumour size with delivered near-infrared (NIR) dye. The nanoparticle will be surface functionalized with novel aptamers for improved targeting. The design can therefore be separated into four parts from outside to inside: aptamerbased targeting system, a pH-responsive charge reversible coating layer, nanoparticle-based NIR non-invasive imaging, and nanoparticle transfected siRNA for alveolar RMS (aRMS) specific gene knockdown. Aptamers were chosen as the targeting ligand for the nanosystem since compared to antibodies, they are much smaller, have higher affinity and specificity, and lower immunogenicity. Furthermore, they are easier and cheaper to manufacture. The binding target for aptamer generation was Neural EGF-Like 1 (NELL1) chosen based on validation results which suggested overexpression on the surface of the target aRMS cells. Oligos were isolated, and their binding affinity was quantified using Surface Plasmon Resonance (SPR). The selected NELL1 aptamer could locate and firmly bind to the NELL1 on the surface of aRMS cells, which could therefore potentially minimize off-target effects. The aptamer was attached to a synthesized pH-activated charge reversible PMPC-PDPA@PEG- 2 b-PAMA/DMMA copolymer layer. The PEG-b-PAMA/DMMA within the copolymer was able to efficiently convert the charge from negative (under physiological pH) to positive (under acidic pH), and the PMPC-PDPA could revert from a hydrophilic to a hydrophobic state after being activated by the acidic environment of tumours and subcellular organelles (endosomes & lysosomes). The enhanced tissue accumulation and cellular uptake could be achieved with increased adsorption onto negatively charged cellular membranes. For NIR imaging, IR820 NIR dye was loaded into the mesoporous silica nanoparticles core to enable non-invasive monitoring of the uptake and accumulation of the nanosystem during treatment. Compared with free IR820 dye which could only rely on passive diffusion, the IR820- loaded MSNPs were found to have significant improvements from the perspectives of cellular uptake, accumulation amount and penetration depth within both 2D and 3D cell models. Two siRNA targeting options were selected and validated for aRMS gene therapy: (i) Human αB-crystallin (CRYAB) & Heat Shock Protein Family B (Small) Member 2 (HSPB2) double knockout, and (ii) Keratin 17 (KRT17) knockdown. Through linking the validated siRNAs onto the MSNPs surface with the disulfide bound which could only be cleavaged by the high concentration of GSH within RMS cells, it has been found to have a high degree of RNase I degradation prevention, and a significant enhanced transfection efficiency. The total GSH levels within RD and RH30 cells were found to be 5.08 and 9.28 times higher than that of control fibroblast cells. Compared with the commercially available DharmaFect transfection reagent, the nanoparticles transfected siRNA has shown a similar or better level of gene knockdown; with a significant suppression of aRMS cells proliferation and mobility. Overall, the constructed nanosystem could provide another route for tackling aRMS in a more effective way, with fewer side-effects.

Zhengguo Qiu,1 Qianhe Shen,2 Chao Jiang,3 Li Yao,4 Xiaopeng Sun,5 Jing Li,6 Chongzhen Duan,1 Rui Li,1 Xiuli Li,1 Subash CB Gopinath,7,8 Periasamy Anbu,9 Thangavel Lakshmipriya,8 Xu Li10 1Department of Anesthesiology, Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine, Xianyang, Shaanxi, 712000, People’s Republic of China; 2Department of Anesthesiology, Xi’an GemFlower Changqing Hospital, Xi’an, Shaanxi, 710200, People’s Republic of China; 3The Third Department of Neurology, The Second Affiliated Hospital of Xi’an Medical University, Xi’an, Shaanxi, 710038, People’s Republic of China; 4Department of Neurology, The Hospital of Xidian Group, Xi’an, Shaanxi, 710077, People’s Republic of China; 5Department of Otolaryngology, The Second Affiliated Hospital of Xi’an Medical University, Xi’an, Shaanxi, 710038, People’s Republic of China; 6Department of Anesthesiology, The Second Affiliated Hospital of Xi’an Medical University, Xi’an, Shaanxi, 710038, People’s Republic of China; 7Faculty of Chemical Engineering Technology, Universiti Malaysia Perlis (UniMAP), Arau, Perlis, 02600, Malaysia; 8Institute of Nano Electronic Engineering, Universiti Malaysia Perlis (UniMAP), Kangar, 01000, Malaysia; 9Department of Biological Engineering, College of Engineering, Inha University, Incheon, 402-751, Republic of Korea; 10Department of Surgery and Anesthesiology, Xi’an Hospital of Traditional Chinese Medicine, Xi’an, Shaanxi, People’s Republic of ChinaCorrespondence: Xu Li Email qzg2020@sina.comBackground: Alzheimer’s disease (AD) is a neurodegenerative chronic disorder that causes dementia and problems in thinking, cognitive impairment and behavioral changes. Amyloid-beta (Aβ) is a peptide involved in AD progression, and a high level of Aβ is highly correlated with severe AD. Identifying and quantifying Aβ levels helps in the early treatment of AD and reduces the factors associated with AD.Materials and Methods: This research introduced a dual probe detection system involving aptamers and antibodies to identify Aβ. Aptamers and antibodies were attached to the gold (Au) urchin and hybrid on the carbon nanohorn-modified surface. The nanohorn was immobilized on the sensor surface by using an amine linker, and then a Au urchin dual probe was immobilized.Results: This dual probe-modified surface enhanced the current flow during Aβ detection compared with the surface with antibody as the probe. This dual probe interacted with higher numbers of Aβ peptides and reached the detection limit at 10 fM with R2=0.992. Furthermore, control experiments with nonimmune antibodies, complementary aptamer sequences and control proteins did not display the current responses, indicating the specific detection of Aβ.Conclusion: Aβ-spiked artificial cerebrospinal fluid showed a similar response to current changes, confirming the selective identification of Aβ.Keywords: amyloid-beta, biomarker, nanomaterial, aptamer, antibody

Jianyuan Song,1 Cuicui Duan,2 Wangyu Cai,1 Wei Wu,1 Houyi Lv,2 Jian Xu1 1The Fourth Affiliated Hospital, Zhejiang University School of Medicine, Yiwu, Zhejiang 322000, People’s Republic of China; 2Department of Assisted Reproduction, Women’s Hospital School of Medicine Zhejiang University, Hangzhou, Zhejiang 310006, People’s Republic of ChinaCorrespondence: Jian XuThe Fourth Affiliated Hospital, Zhejiang University School of Medicine, Yiwu, Zhejiang 322000, People’s Republic of ChinaEmail xuj@zju.edu.cnPurpose: Gonadotrophin releasing hormone agonist (GnRH-a) is widely used for pituitary down-regulation and recruiting more follicles in assisted reproduction. However, no information is available on its value for patients with thin endometrial thickness.Patients and Methods: This was a retrospective cohort study of 302 patients with endometrium < 8 mm undergoing fresh embryo transfer at a fertility center of a university hospital from January 2016 and December 2018. In 148 cycles of the GnRH-a prolonged protocol, one depot of 3.75 mg GnRH-a was injected on day 2 of the menstrual cycle, while in 154 cycles of the short GnRH-a long protocol, 0.1 mg of GnRH-a was injected daily from the mid-luteal phase. The live birth rate and clinical pregnancy rate were compared between the two groups. Other outcome measures included the implantation rate, miscarriage rate, and characteristics of stimulation procedures.Results: Live birth rates and clinical pregnancy rates were significantly higher in the GnRH-a prolonged protocol group than in the other group (36.5% vs 20.8%, P=0.002; 43.9% vs 28.2%, P=0.006, respectively). The live birth rate was significantly increased in the prolonged protocol group (crude OR: 2.190, 95% CI: 1.311, 3.660; adjusted OR: 2.458, 95% CI: 1.430, 4.224) compared with that in the reference group. The implantation rate of the former group was also significantly higher than that of the latter group (35.4% vs 15.9%, P=0.000). There was no significant difference in miscarriage rates between the two protocols. In terms of stimulation procedures, the GnRH-a prolonged protocol group required significantly higher Gn time (10.9 vs 9.5 days, P=0.000) and Gn consumption (2625.0 vs 2047.5 IU, P=0.000) than the short GnRH-a long protocol group.Conclusion: The GnRH-a prolonged protocol in fresh embryo transfer cycles yielded better clinical outcomes of patients with thin endometrium than the short GnRH-a long protocol.Keywords: ovarian stimulation, GnRH-a prolonged protocol, thin endometrium, IVF, COH

Honey bees are a model for host-microbial interactions with experimental designs evolving towards conventionalized worker bees. Research on gut microbiome transmission and assembly has examined only a fraction of factors associated with the colony and hive environment. Here, we studied the effects of diet and social isolation on tissue-specific bacterial and fungal colonization of the midgut and two key hindgut regions. We found that both treatment factors significantly influenced early hindgut colonization explaining similar proportions of microbiome variation. In agreement with previous work, social interaction with older workers was unnecessary for core hindgut bacterial transmission. Exposure to natural eclosion and fresh stored pollen resulted in gut bacterial communities that were taxonomically and structurally equivalent to those produced in the natural colony setting. Stressed diets of no pollen or autoclaved pollen in social isolation resulted in decreased fungal abundance and bacterial diversity, and atypical microbiome structure and tissue-specific variation of functionally important core bacteria. Without exposure to the active hive environment, the abundance and strain diversity of keystone ileum species Gilliamella apicola was markedly reduced. These changes were associated with significantly larger ileum microbiotas suggesting that extended exposure to the active hive environment plays an antibiotic role in hindgut microbiome establishment. We conclude that core hindgut microbiome transmission is facultative horizontal with 5 of 6 core hindgut species readily acquired from the built hive structure and natural diet. Our findings contribute novel insights into factors influencing assembly and maintenance of honey bee gut microbiota and facilitate future experimental designs.

Junling Yang,1 Zaijun Hao,1 Wei Li,1 Caiping Duan,1 Xiujuan Fan,1 Jing Xin,1 Chunguang Ren2 1Department of Anesthesiology, Ordos Central Hospital, Ordos, People’s Republic of China; 2Department of Anesthesiology, Liaocheng People’s Hospital, Liaocheng, Shandong, People’s Republic of ChinaCorrespondence: Chunguang Ren Email weiyichunguang@163.comBackground: Although video-assisted thoracoscopic surgery (VATS) is increasingly used, the optimal analgesia strategy is still unknown. We explore the efficacy and safety of preemptive ultrasound-guided paravertebral block (PVB) combined with parecoxib during VATS.Methods: Seventy-four patients were divided into two groups. PVBs were performed before anesthesia induction under real-time ultrasound guidance. Visual analog scale (VAS) score with coughing at 48 h after surgery, postoperative sufentanil consumption and level of sedation (LOS) at 1, 4, 8, 12, 24, and 48 h postoperatively, intraoperative hemodynamics, satisfaction scores of patients and surgeons, remedial measures, time to chest tube removed and mobilization, adverse effects and hospital stay length were recorded. We also recorded inflammatory markers, respiratory function and the prevalence of chronic pain after surgery.Results: VAS scores at rest and with coughing during the first 24 h after surgery were significantly lower in the P group (P< 0.05). Consumption of sevoflurane, remifentanil, and dexmedetomidine was all significantly reduced in the P group (P< 0.05). The consumption of sufentanil within 48 h after surgery, time to first dose and total dose of rescue ketorolac was significantly lower in the P group (P< 0.05). The FEV1/FVC ratio was significantly higher in the P group at 1 and 3 d after surgery (P< 0.05). Times to chest tube removal and mobilization were significantly shorter in the P group (P< 0.05). Compared with the C group, the level of both ACTH and cortisol was significantly reduced in the P group at 1 and 3 d after surgery (P< 0.05).Conclusion: PVB combined with parecoxib was associated with better pain relief, decreased sufentanil and ketorolac consumption, less hemodynamic instability, and a lower surgery-related stress response. However, the incidences of chronic pain 3 and 6 months after surgery and the risk of complication except urinary retention were not significantly different between groups.Keywords: paravertebral block, parecoxib, video-assisted thoracic surgery, preemptive analgesia

Simple Summary Honey bees are vital to the agriculture of the world, but like all managed organisms, disease control has become challenging due to the overuse and misuse of antibiotics. Alternate solutions with potential to control disease include natural compounds and probiotic supplements. Probiotic supplements in honey bees have been praised by industry, but studies applying probiotics to honey bee larval disease are lacking and technically challenging. In this study we tested the effectiveness of a demonstrated probiotic (Parasacharribacter apium strain C6) to mitigate a damaging larval disease called European Foul Brood (EFB). Based on a controlled laboratory study and two separate trials, the probiotic had no effect on EFB disease. The control groups performed as expected, validating the very sensitive lab procedure used to artificially rear honey bee larvae. Surprisingly, the probiotic provided no survival benefit to larvae in the absence of disease, contradicting past results. We discuss the difficult technique of larval rearing in the laboratory with reference to an improved experimental design introducing disease agents and potential remedies. In summary, our findings indicate that the representation of honey bee health and disease in the laboratory setting requires repeatable validation with reference to rigorous control and natural colony context. Abstract European honey bees (Apis mellifera Linnaeus) are beneficial insects that provide essential pollination services for agriculture and ecosystems worldwide. Modern commercial beekeeping is plagued by a variety of pathogenic and environmental stressors often confounding attempts to understand colony loss. European foulbrood (EFB) is considered a larval-specific disease whose causative agent, Melissococcus plutonius, has received limited attention due to methodological challenges in the field and laboratory. Here, we improve the experimental and informational context of larval disease with the end goal of developing an EFB management strategy. We sequenced the bacterial microbiota associated with larval disease transmission, isolated a variety of M. plutonius strains, determined their virulence against larvae in vitro, and explored the potential for probiotic treatment of EFB disease. The larval microbiota was a low diversity environment similar to honey, while worker mouthparts and stored pollen contained significantly greater bacterial diversity. Virulence of M. plutonius against larvae varied markedly by strain and inoculant concentration. Our chosen probiotic, Parasaccharibacter apium strain C6, did not improve larval survival when introduced alone, or in combination with a virulent EFB strain. We discuss the importance of positive and negative controls for in vitro studies of the larval microbiome and disease.

Background In social insects, identical genotypes can show extreme lifespan variation providing a unique perspective on age-associated microbial succession. In honey bees, short- and long-lived host phenotypes are polarized by a suite of age-associated factors including hormones, nutrition, immune senescence, and oxidative stress. Similar to other model organisms, the aging gut microbiota of short-lived (worker) honey bees accrue Proteobacteria and are depleted of Lactobacillus and Bifidobacterium, consistent with a suite of host senescence markers. In contrast, long-lived (queen) honey bees maintain youthful cellular function with much lower expression of oxidative stress genes, suggesting a very different host environment for age-associated microbial succession. Results We sequenced the microbiota of 63 honey bee queens exploring two chronological ages and four alimentary tract niches. To control for genetic and environmental variation, we quantified carbonyl accumulation in queen fat body tissue as a proxy for biological aging. We compared our results to the age-specific microbial succession of worker guts. Accounting for queen source variation, two or more bacterial species per niche differed significantly by queen age. Biological aging in queens was correlated with microbiota composition highlighting the relationship of microbiota with oxidative stress. Queens and workers shared many major gut bacterial species, but differ markedly in community structure and age succession. In stark contrast to aging workers, carbonyl accumulation in queens was significantly associated with increased Lactobacillus and Bifidobacterium and depletion of various Proteobacteria. Conclusions We present a model system linking changes in gut microbiota to diet and longevity, two of the most confounding variables in human microbiota research. The pattern of age-associated succession in the queen microbiota is largely the reverse of that demonstrated for workers. The guts of short-lived worker phenotypes are progressively dominated by three major Proteobacteria, but these same species were sparse or significantly depleted in long-lived queen phenotypes. More broadly, age-related changes in the honey bee microbiota reflect the regulatory anatomy of reproductive host metabolism. Our synthesis suggests that the evolution of colony-level reproductive physiology formed the context for host-microbial interactions and age-related succession of honey bee microbiota. Electronic supplementary material The online version of this article (10.1186/s40168-018-0489-1) contains supplementary material, which is available to authorized users.

Youyi Dai,1 Huaxin Duan,2,3 Chaojun Duan,4 Hong Zhu,1 Rongrong Zhou,1 Haiping Pei,5 Liangfang Shen1 1Department of Oncology, Xiangya Hospital, Central South University, 2Department of Oncology, Hunan Provincial People’s Hospital, 3Department of Oncology, The First Affiliated Hospital of Hunan Normal University, 4Institute of Medical Sciences, Xiangya Hospital, Central South University, 5Department of General Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, People’sRepublic of China Abstract: Colorectal cancer (CRC) has become a major public health problem, ranking as the third most common type of cancer. Our previous study has revealed that TCF21 is frequently silenced by promoter hypermethylation in both CRC cell lines and primary CRC, with TCF21 methylation being significantly correlated with lymph node invasion. In this study, we further analyze the expression of TCF21 in CRC tissues and investigate the role of TCF21 in CRC invitro and in vivo. We also explore the possible pathway regulated by TCF21. We thus demonstrate that decreased levels of TCF21 are associated with the pathological stage, clinical stage and lymph node metastasis, indicating a poor prognosis in CRC patients; overexpression of TCF21 inhibits cell proliferation, migration and invasion in the colorectal cell lines HCT116 and HT29. Furthermore, TCF21 functions as a tumor suppressor probably through inactivation of PI3K/AKT signaling and inhibition of MMPs. Our results suggest that enhancement of TCF21 levels may be a potential strategy to facilitate the prevention and treatment of CRC in the clinic. Keywords: colorectal cancer, TCF21, prognosis, PI3K/AKT signaling, MMPs

Honey bee colony performance and health are intimately linked to the foraging environment. Recent evidence suggests that the US Conservation Reserve Program (CRP) has a positive impact on environmental suitability for supporting honey bee apiaries. However, relatively little is known about the influence of habitat conservation efforts on honey bee colony health. Identifying specific factors that influence bee health at the colony level incorporates longitudinal monitoring of physiology across diverse environments. Using a pooled-sampling method to overcome individual variation, we monitored colony-level molecular biomarkers during critical pre- and post-winter time points. Major categories of colony health (nutrition, oxidative stress resistance, and immunity) were impacted by apiary site. In general, apiaries within foraging distance of CRP lands showed improved performance and higher gene expression of vitellogenin (vg), a nutritionally regulated protein with central storage and regulatory functions. Mirroring vg levels, gene transcripts encoding antioxidant enzymes and immune-related proteins were typically higher in colonies exposed to CRP environments. Our study highlights the potential of CRP lands to improve pollinator health and the utility of colony-level molecular diagnostics to assess environmental suitability for honey bees.

© 2019 Di Wu et al. This network meta-analysis addresses the need for evidence-based best-practice treatment regimens for HER2-positive breast cancer. We compared the relative efficacy and tolerability of currently available HER2-positive neoadjuvant immunotherapy regimens based on systematic searches of available randomized controlled trials (RCTs) data. Based on intention-to-treat principle, pathological complete response (pCR), overall serious adverse events (SAEs), and breast-conserving surgery (BCS) rate were analyzed using random-effect, Bayesian network meta-analysis, and standard pairwise meta-analysis. 16 RCTs (3868 patients) were included. Analyzed treatment regimens were as follows: chemotherapy+trastuzumab+pertuzumab (CTP), trastuzumab emtansine+pertuzumab (MP), chemotherapy+trastuzumab (CT), chemotherapy+pertuzumab (CP), trastuzumab+pertuzumab (TP), chemotherapy+trastuzumab+lapatinib (CTL), and chemotherapy+lapatinib (CL), and chemotherapy (C) alone. We found that, for the chance of achieving pCR, CTP was ranked first (SUCRA: 97%), followed by CTL, MP, and CT (SUCRA: 80%, 75%, and 55%, resp.). MP provided the safest regimen (SUCRA: 97%), then TP, C, and TPC (SUCRA: 82%, 76%, and 47%, resp.). CTL proved the most toxic therapy (SUCRA: 7%). No significant difference between neoadjuvant regimens was identified for BCS. Hormone receptor status did not impact ORs for pCR in any regimen. In conclusion, our findings support CTP as the optimum neoadjuvant regimen for HER2-positive breast cancer, with the best pCR and acceptable toxicity compared with CT. MP provides a therapeutic option for patients with poor performance status.

BackgroundIn social insects, identical genotypes can show extreme lifespan variation providing a unique perspective on age-associated microbial succession. In honey bees, short and long-lived host phenotypes are polarized by a suite of age-associated factors including hormones, nutrition, immune senescence and oxidative stress. Similar to other model organisms, the aging gut microbiota of short-lived (worker) honey bees accrue Proteobacteria and are depleted of Lactobacillus and Bifidobacterium, consistent with a suite of host senescence markers. In contrast, long-lived (queen) honey bees maintain youthful cellular function without expressing oxidative stress genes, suggesting a very different host environment for age-associated microbial succession.ResultsWe sequenced the microbiota of 63 honey bee queens exploring two chronological ages and four alimentary tract niches. To control for individual variation we quantified carbonyl accumulation in queen fat body tissue as a proxy for biological aging. We compared our results to the age-specific microbial succession of worker guts. Accounting for queen source variation, two or more bacterial species per niche differed significantly by queen age. Biological aging in queens was correlated with microbiota composition highlighting the relationship of microbiota with oxidative stress. Queens and workers shared many major gut bacterial species, but differ markedly in community structure and age succession. In stark contrast to aging workers, carbonyl accumulation in queens was significantly associated with increased Lactobacillus and Bifidobacterium and depletion of various Proteobacteria.ConclusionsWe present a model system linking changes in gut microbiota to diet and longevity, two of the most confounding variables in human microbiota research. As described for other model systems, metabolic changes associated with diet and host longevity correspond to the changing microbiota. The pattern of age-associated succession in the queen microbiota is largely the reverse of that demonstrated for workers. The guts of short-lived worker phenotypes are progressively dominated by three major Proteobacteria, but these same species were sparse or significantly depleted in long-lived queen phenotypes. More broadly, our results suggest that lifespan evolution formed the context for host-microbial interactions and age-related succession of honey bee microbiota.

Honey bee colony nutritional ecology relies on the acquisition and assimilation of floral resources across a landscape with changing forage conditions. Here, we examined the impact of nutrition and queen age on colony health across extended periods of reduced forage in a southern climate. We measured conventional hive metrics as well as colony-level gene expression of eight immune-related genes and three recently identified homologs of vitellogenin (vg), a storage glycolipoprotein central to colony nutritional state, immunity, oxidative stress resistance and life span regulation. Across three apiary sites, concurrent longitudinal changes in colony-level gene expression and nutritional state reflected the production of diutinus (winter) bees physiologically altered for long-term nutrient storage. Brood production by young queens was significantly greater than that of old queens, and was augmented by feeding colonies supplemental pollen. Expression analyses of recently identified vg homologs (vg-like-A, -B, and -C) revealed distinct patterns that correlated with colony performance, phenology, and immune-related gene transcript levels. Our findings provide new insights into dynamics underlying managed colony performance on a large scale. Colony-level, molecular physiological profiling is a promising approach to effectively identify factors influencing honey bee health in future landscape and nutrition studies.

Carbohydrate-active enzymes play an important role in the honey bee (Apis mellifera) due to its dietary specialization on plant-based nutrition. Secretory glycoside hydrolases (GHs) produced in worker head glands aid in the processing of floral nectar into honey and are expressed in accordance with age-based division of labor. Pollen utilization by the honey bee has been investigated in considerable detail, but little is known about the metabolic fate of indigestible carbohydrates and glycosides in pollen biomass. Here, we demonstrate that pollen consumption stimulates the hydrolysis of sugars that are toxic to the bee (xylose, arabinose, mannose). GHs produced in the head accumulate in the midgut and persist in the hindgut that harbors a core microbial community composed of approximately 108 bacterial cells. Pollen consumption significantly impacted total and specific bacterial abundance in the digestive tract. Bacterial isolates representing major fermentative gut phylotypes exhibited primarily membrane-bound GH activities that may function in tandem with soluble host enzymes retained in the hindgut. Additionally, we found that plant-originating β-galactosidase activity in pollen may be sufficient, in some cases, for probable physiological activity in the gut. These findings emphasize the potential relative contributions of host, bacteria, and pollen enzyme activities to carbohydrate breakdown, which may be tied to gut microbiome dynamics and associated host nutrition.

Leukemic lymphoblasts within different immunophenotypic populations possess stem cell properties. However, whether or not the self-renewal program is retained from stem cells or conferred on progenitors by leukemogenic molecules remains unknown. We have addressed the issue in the context of TEL-AML1-associated acute lymphoblastic leukemia (ALL) by profiling a refined program edited from genes essential for self-renewal of hematopoietic stem cells and B-cell development. Bioinformatic analysis shows that ALL populations are loosely clustered and close to the normal population that contains stem and primitive progenitor cells. This finding indicates that immunophenotypes do not reflect maturation stages in ALL and that the self-renewal program may be retained from stem cells. Results of assessing 'first hit' function of TEL-AML1 in different populations of normal cells demonstrate the molecular model. Therefore, the current study shows a leukemogenic scenario of human ALL in which programs of stem cells are sustained in distinct fractions by leukemogenic mutations.

Bao Liu,1 Larry Z Liu,2,3 Jianwei Xuan,2 Man Luo,4 Yansheng Li,5 Chaohui Duan,6 Hongqin Cheng,7 Xiaohui Yang81School of Public Health, Fudan University, Shanghai, People’s Republic of China; 2Pfizer Inc, New York, NY, USA; 3Weill Medical College of Cornell University, New York, NY, USA; 4Zhongshan Hospital, Fudan University, Shanghai, People’s Republic of China; 5Renji Hospital, Shanghai Jiaotong University, Shanghai, People’s Republic of China; 6The Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, People’s Republic of China; 7Xuanwu Hospital, Beijing, People’s Republic of China; 8Beijing Anzhen Hospital, Capital University of Medical Science, Beijing, People’s Republic of ChinaBackground: Atrial fibrillation (AF) is associated with an increased risk of stroke. This study assessed treatment patterns associated with stroke prevention among patients with AF in three major cities of the People’s Republic of China.Methods: A random sample of 2,862 medical charts for patients with AF at six tertiary hospitals located in Beijing, Shanghai, and Guangzhou between 2003 and 2008 were reviewed. Patient demographics, clinical characteristics, and treatment patterns were extracted from medical charts. Antithrombotic regimens included antiplatelets, anticoagulants, and a combination of both. Descriptive analyses were performed to summarize basic antithrombotic patterns. A logistic regression model examined demographic and clinical factors associated with antithrombotic treatment patterns.Results: Of the patient sample, 55% were male, the average age was 72 years (49% ≥75 years), 15% had valvular AF, 78% had nonvalvular AF, and the remainder had unspecified AF. CHADS2 scores ≥2 were reported for 53% of patients. Antithrombotic treatment was not received by 17% of patients during hospitalization, and 66% did not receive warfarin. Among patients with valvular or nonvalvular AF, 33%, 30%, and 20% received antiplatelet, anticoagulation, and antiplatelet plus anticoagulation treatments, respectively. For patients with CHADS2 scores of 0, 1, 2, 3, and ≥4, 52%, 42%, 28%, 21%, and 21%, respectively, were treated with warfarin. Predictors of no antithrombotic treatment included age and hospital location.Conclusion: Anticoagulation therapy was underused in Chinese patients with AF. Antithrombotic treatment was not associated with stroke risk. Further studies need to examine the clinical consequences of various antithrombotic treatment patterns in Chinese patients with AF.Keywords: atrial fibrillation, stroke prevention, antithrombotic treatment, China

Feihu Wang1, Yuxuan Chen2, Dianrui Zhang1, Qiang Zhang3, Dandan Zheng1, Leilei Hao1, Yue Liu1, Cunxian Duan1, Lejiao Jia1, Guangpu Liu11Department of Pharmaceutics, College of Pharmacy, Shandong University, Jinan, People’s Republic of China; 2Department of Pharmacy, Shenzhou Hospital, Shenyang, People’s Republic of China; 3State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, People’s Republic of ChinaBackground: A critical disadvantage for successful chemotherapy with paclitaxel (PTX) is its nontargeting nature to cancer cells. Folic acid has been employed as a targeting ligand of various anticancer agents to increase their cellular uptake within target cells since the folate receptor is overexpressed on the surface of such tumor cells. In this study, a novel biodegradable deoxycholic acid-O-carboxymethylated chitosan–folic acid conjugate (DOMC-FA) was used to form micelles for encapsulating the anticancer drug PTX.Methods and results: The drug-loading efficiency, encapsulation efficiency, in vitro drug release and physicochemical properties of PTX-loaded micelles were investigated in detail. In vitro cell culture studies were carried out in MCF-7 cells, a human breast carcinoma cell line, with folate receptor overexpressed on its surface. An increased level of uptake of folate-conjugated micelles compared to plain micelles in MCF-7 cells was observed, and the enhanced uptake of folate-micelles mainly on account of the effective process of folate receptor-mediated endocytosis. The MTT assay, morphological changes, and apoptosis test implied that the folate-conjugated micelles enhanced the cell death by folate-mediated active internalization, and the cytotoxicity of the FA-micellar PTX (DOMC-FA/PTX) to cancer cells was much higher than micelles without folate (DOMC/PTX) or the commercially available injectable preparation of PTX (Taxol).Conclusion: Results indicate that the PTX-loaded DOMC-FA micelle is a successful anticancer-targeted drug-delivery system for effective cancer chemotherapy.Keywords: paclitaxel, folate, polymeric micelles, targeted delivery

OBJECTIVES To investigate whether angiotensin II (AII) receptor antagonism decreases the inflammation and oedema in acute murine experimental autoimmune cystitis (EAC), as interstitial cystitis (IC) might have an autoimmune component and AII has been implicated in autoimmune-mediated vascular congestion, oedema and scarring. MATERIALS AND METHODS Female Balb/cAN mice were divided into three treatment groups (eight in each group) that were autoimmunized with bladder homogenate to induce EAC. One group received an AII type 1 receptor (AT1) antagonist, one group an AII type 2 receptor (AT2) antagonist, and one group remained untreated (EAC). A control and sham-injected group were also included. After 10 weeks, bladders were removed, sectioned, and stained with haematoxylin and eosin. RESULTS Grossly, there was no thickening or adhesions in the bladders of the control or sham-injected mice. In five of seven surviving EAC bladders, there were dense adhesions to surrounding peritoneal structures. There were also adhesions and bladder thickening in all of the AT2 antagonist-treated mice (though in a milder form) but in only two of seven surviving AT1 antagonist-treated mice. There was no inflammation or oedema in the sham and control groups. All the EAC bladders were inflamed, with submucosal oedema and urothelial detachment from the lamina propria. In the AT1 antagonist-treated mice there was no inflammation or oedema. By contrast, all AT2 antagonist-treated mice had moderate inflammation and minor detachment of the urothelium from the lamina propria. CONCLUSIONS AT1 receptor blockade ameliorated the inflammatory infiltration, submucosal oedema, and urothelial detachment associated with EAC in mice. This was achieved to a lesser extent by AT2 receptor blockade. If some patients with IC have a pathophysiology similar to that of EAC mice, there might be potential benefit from AII receptor blockade.

Ischemia/reperfusion injury is a leading cause of renal damage and antisense gene therapy has been shown to ameliorate its effects. However, this approach has been limited by current delivery methods that require high concentrations of intravenous nucleic acids lacking specificity for targeting tissues. To overcome these limitations we developed a novel murine partial nephrectomy model to evaluate polyethylene-glycol (PEG) hydrogel tissue sealant as a topical oligonucleotide delivery system.A total of 18 male C57BL/6 mice underwent left partial nephrectomy with vascular occlusion. Hydrogel primer and then sealant were applied to the cut surface and photopolymerized. Using this method 16 additional mice received hydrogel primer mixed with Cy5 labeled fluorescent oligonucleotide (10 to 100 microg). Kidneys were harvested at various time points and assessed for oligonucleotide penetration using fluorescence microscopy.A survival rate of 100% (34 subjects) was obtained using this mouse model of partial nephrectomy. PEG hydrogel provided adequate protection against renal hematoma and intraperitoneal blood. Fluorescent images revealed that 50 microg was the minimum dose resulting in complete progressive cellular penetration with time. In addition to direct diffusion from the application site, movement of oligonucleotide through the subcapsular space into the cortex was an observed mechanism of distribution.A murine partial nephrectomy model was successfully created using PEG hydrogel. In addition to achieving hemostasis, hydrogel served as a successful depot for delivering oligonucleotides throughout the kidney.

UTokyo Research掲載「動物の成長や細胞の増殖の新しい調節機構を発見」 URI: http://www.u-tokyo.ac.jp/ja/utokyo-research/research-news/novel-regulatory-mechanisms-of-animal-growth-and-cell-proliferation.html, UTokyo Research "Novel regulatory mechanisms of animal growth and cell proliferation" URI: http://www.u-tokyo.ac.jp/en/utokyo-research/research-news/novel-regulatory-mechanisms-of-animal-growth-and-cell-proliferation.html

The products and the related gene expression of flavonoid metabolism were examined in the skin and pulp of Yan73 (Vtis vinifera L. cv.) grapes, a teinturier variety cultivated in China, and the data were compared with those in the skin and pulp of Cabernet Sauvignon (Vitis vinifera L. cv.) grapes, a well-known red variety. The results showed that, in comparison with the skins of Cabernet Sauvignon, the skins of Yan73 berries were characterised by fewer types of flavonols and anthocyanins, a higher level of flavonoids, and a much lower percentage of 3'-hydroxylated flavonols. Flavonoid metabolism was also present in the pulp of Yan73, and the percentages of 3'-hydroxylated flavonols and 3'-hydroxylated flavan-3-ols were significantly higher than the percentages of their corresponding 3',5'-hydroxylated forms. Only flavan-3-ols were found in the pulp of Cabernet Sauvignon. The expression of VvmybAl exhibited a co-ordination with the accumulation of anthocyanins in the pulp of the teinturier cultivar, which showed great similarity to that in the skin. A good correlation was found between the expression of VvF3'H and 3'-hydroxylated flavonoids, as well as between the expression of VvF3'5'H and 3',5'-hydroxylated flavonoids in the skin and pulp of the grape berries. It is suggested that tissue-specific accumulation of flavonoids in grape berries is related to the transcriptional expression of VvF3'H and VvF3'5'H.

Proanthocyanidins in grape berries are synthesised mainly before véraison, and very little attention is paid to the evolution of proanthocyanidins (PAs) in grapes from véraison to harvest. The present study focused on the changes of flavan-3-ols with different degrees of polymerisation in grape skins and the difference in proanthocyandin composition of 'Shiraz', 'Cabernet Sauvignon' and 'Marselan' grapes (Vitis vinifera L.). The results show that the content of flavan-3-ols, the percentage of prodelphinidins (%P) and mean degree polymerisation (mDP) found in 'Cabernet Sauvignon' berry skins at post-véraison were higher than those in 'Shiraz' and 'Marselan' skins. Only monomelic, dimeric, trimeric and polymeric flavan-3-ols were detected in the three grape cultivars. Polymers with more than tenfold flavan-3-ol units accounted for a relatively high proportion in grape berry skins, and the content in the three cultivars declined continuously during ripening. Principal component analysis showed that proanthocyanidin content, composition and mDP at grape harvest stage depended strongly on grape cultivar. This study provides some useful information for understanding the accumulation of PAs during berry maturation and this information can be used to improve wine quality.

This study demonstrates inducible transgenic expression in the exceptionally short-lived turquoise killifish Nothobranchius furzeri, which is a useful vertebrate model for ageing research. Transgenic N. furzeri bearing a green fluorescent protein (GFP), containing construct under the control of a heat shock protein 70 promoter were generated, heat shock-induced and reversible GFP expression was demonstrated, and germline transmission of the transgene to the F1 and F2 generations was achieved. The availability of this inducible transgenic expression system will make the study of ageing-related antagonistically pleiotropic genes possible using this unique vertebrate model organism.

The separation performance of a raw coal sample from South Africa based on the dense gas-solid fluidized bed beneficiation technique was investigated. Analysis showed that the raw coal sample distributes mainly in the -50+6 mm size range with the -1.6 g/cm³ density fraction accounting for 78.20% of the cumulative mass distribution. A flow sheet of the sequential coal beneficiation technique was introduced. The experimental and simulation results of the hydromechanical and density distribution stability indicate that the dense gas-solid fluidized bed could provide a stable separation environment with uniform density fluctuations under suitable operating conditions. With the application of this technique on the raw coal sample, the separation results show that three products, consisting of 73.25% clean coal with an ash content of 9.59%, 11.31% middlings, and 15.44% gangue were effectively separated at two different densities of 1.82 g/cm³ and 1.57 g/cm³. This study provides an effective flow sheet for achieving the high-efficiency separation performance on raw coal from arid areas.

By means of two different parametrizations of quark energy loss and the nuclear parton distributions determined only with lepton-nuclear deep inelastic scattering experimental data, a leading order phenomenological analysis is performed on the nuclear Drell-Yan differential cross section ratios as a function of the quark momentum fraction in the beam proton and target nuclei for E772 experimental data. It is shown that there is the quark energy loss effect in nuclear Drell-Yan process apart from the nuclear effects on the parton distribution as in deep inelastic scattering. The uncertainties due to quark energy loss effect is quantified on determining nuclear sea quark distribution by using nuclear Drell-Yan data. It is found that the quark energy loss effect on nuclear Drell-Yan cross section ratios make greater with the increase of quark momentum fraction in the target nuclei. The uncertainties from quark energy loss become bigger as the nucleus A come to be heavier. The Drell-Yan data on proton incident middle and heavy nuclei versus deuterium would result in an overestimate for nuclear modifications on sea quark distribution functions with neglecting the quark energy loss. Our results are hoped to provide good directional information on the magnitude and form of nuclear modifications on sea quark distribution functions by means of the nuclear Drell-Yan experimental data., Comment: 16 pages, 3 figures

Fluorescence lifetimes of Eu$^{2+}$ doped in CaF$_2$ are measured at various temperatures between 4K and 450K. The lifetime increases with between 100K and 300K, in contrast to the usual lifetime-temperature dependence. At higher or lower temperatures the lifetime decreases with increasing temperature. The phenomenon is explained by simulation of the energy levels and lifetimes of low-lying $4f^65d$ states involved in the fluorescence., Comment: 10 pages, 2 figures, to appear on Journal of Alloys and Compounds

We present molecular dynamics simulations of solid NaNO$_2$ using pair potentials with the rigid-ion model. The crystal potential surface is calculated by using an \emph{a priori} method which integrates the \emph{ab initio} calculations with the Gordon-Kim electron gas theory. This approach is carefully examined by using different population analysis methods and comparing the intermolecular interactions resulting from this approach with those from the \emph{ab initio} Hartree-Fock calculations. Our numerics shows that the ferroelectric-paraelectric phase transition in solid NaNO$_2$ is triggered by rotation of the nitrite ions around the crystallographical c axis, in agreement with recent X-ray experiments [Gohda \textit{et al.}, Phys. Rev. B \textbf{63}, 14101 (2000)]. The crystal-field effects on the nitrite ion are also addressed. Remarkable internal charge-transfer effect is found., Comment: RevTeX 4.0, 11 figures

We present molecular dynamics simulations of solid NaNO$_2$ using pair potentials with the rigid-ion model. The crystal potential surface is calculated by using an \emph{a priori} method which integrates the \emph{ab initio} calculations with the Gordon-Kim electron gas theory. This approach is carefully examined by using different population analysis methods and comparing the intermolecular interactions resulting from this approach with those from the \emph{ab initio} Hartree-Fock calculations. Our numerics shows that the ferroelectric-paraelectric phase transition in solid NaNO$_2$ is triggered by rotation of the nitrite ions around the crystallographical c axis, in agreement with recent X-ray experiments [Gohda \textit{et al.}, Phys. Rev. B \textbf{63}, 14101 (2000)]. The crystal-field effects on the nitrite ion are also addressed. Remarkable internal charge-transfer effect is found., RevTeX 4.0, 11 figures

Insulin-like growth factor binding protein-5 (IG-FBP-5) is an important modulator of IGF actions. IG-FBP-5 mRNA is abundant in human fibroblasts and is regulated by cAMP. To understand the molecular mechanism underlying this cell type-specific expression and regulation, we isolated the 5'-flanking region of the human IGFBP-5 gene and fused it to a promoter-less reporter plasmid encoding luciferase. Transient transfection of the construct into fibroblasts displayed both constitutive and cAMP-induced promoter activity in an orientation-specific manner. Sequence analysis revealed the existence of distal and proximal consensus AP-2 recognition sites located 5' from the TATA box. Both sequences bound specifically to human AP-2 in vitro by gel shift mobility assay. The possible role of AP-2 was examined by cotransfection of AP-2-deficient HepG2 cells with the IGFBP-5 promoter construct and a human AP-2 expression construct. Cotransfection with AP-2 significantly elevated IGFBP-5 promoter activity. This trans-activation was IGFBP-5 promoter and AP-2 specific. In AP-2 abundant fibroblasts, expression of AP-2B, a dominant-negative inhibitor of AP-2, suppressed IGFBP-5 promoter activity. In HepG2 cells, AP-2B alone had no significant effect, but the AP-2-induced activation of promoter activity was inhibited by AP-2B in a dose-dependent manner. The relative functional importance of the putative AP-2 binding sites was examined using a number of deletion mutants and point mutations. When the first two distal CCCCACCC-like putative AP-2 sites were deleted or mutated, there was no change in AP-2-induced trans-activation. Deletion or mutation of the proximal GCCNNNGGC-like sequences, however, abolished the AP-2-induced activation. These results suggest that AP-2 regulates the IGFBP-5 gene expression through the proximal GCCNNNGGC-like sequences. This AP-2-mediated trans-activation contributes at least in part to the constitutively high expression of IGFBP-5 in fibroblasts and to the cAMP responsiveness of this gene.