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Background Diabetic eye screening programmes have been developed worldwide based on evidence that early detection and treatment of diabetic retinopathy are crucial to preventing sight loss. However, little is known about the decision-making processes and training needs of diabetic retinal graders, particularly in low- and middle-income countries. Objectives To provide data for improving evidence-based diabetic retinopathy training to help novice graders process fundus images more like experts. Subjects/methods This is a mixed-methods qualitative study conducted in southern Vietnam and Northern Ireland. Novice diabetic retinal graders in Vietnam (n = 18) and expert graders in Northern Ireland (n = 5) were selected through a purposive sampling technique. Data were collected from 21st February to 3rd September 2019. The interviewer used neutral prompts during think-aloud sessions to encourage participants to verbalise their thought processes while grading fundus images from anonymised patients, followed by semi-structured interviews. Thematic framework analysis was used to identify themes, supported by illustrative quotes from interviews. Mann–Whitney U tests were used to compare graders’ performance. Results Expert graders used a more systematic approach when grading images, considered all four images per patient and used available software tools such as red-free filters prior to making a decision on management. The most challenging features for novice graders were intra-retinal microvascular abnormalities and new vessels, which were more accurately identified by experts. Conclusion Taking more time to grade fundus images and adopting a protocol-driven “checklist” approach may help novice graders to function more like experts.

A 25-year-old male patient visited the ophthalmology clinic because of upper eye lid ptosis in the right eye, binocular double vision, and light sensitivity. He was diagnosed with a complete third nerve palsy caused by a skull base myofibroma, a rare clinical entity that has not been described before in oculomotor nerve palsy.

An SLC NAND array based in-flash computing core is proposed for enabling vector-matrix multiplications in binarized neural network (BNN) and binary weight network (BWN). Two SLC NAND floating gate (FG) cells in the same string store complementary data to encode binarized weight value of a single synapse for realizing the BNN algorithm. By activating multiple BLs and WLs across block and/or plane levels, the performance of vector-matrix multiplications is significantly accelerated. The system architecture using in-flash computing core for BNN/BWN acceleration is introduced. A wide range of BNN/BWN models can be efficiently mapped to a scalable array of in-flash computing core. An in-flash computing core is able to achieve an estimated peak energy efficiency of 2.56 TOP/s/W for BNN, 292.6 GOP/s/W for 8-b input based BWN by customizing the numbers of word-lines (WLs)/BLs/blocks/planes.

Prokaryotic cell transcriptomics has been limited to mixed or sub-population dynamics and individual cells within heterogeneous populations, which has hampered further understanding of spatiotemporal and stage-specific processes of prokaryotic cells within complex environments. Here we develop a ‘TRANSITomic’ approach to profile transcriptomes of single Burkholderia pseudomallei cells as they transit through host cell infection at defined stages, yielding pathophysiological insights. We find that B. pseudomallei transits through host cells during infection in three observable stages: vacuole entry; cytoplasmic escape and replication; and membrane protrusion, promoting cell-to-cell spread. The B. pseudomallei ‘TRANSITome’ reveals dynamic gene-expression flux during transit in host cells and identifies genes that are required for pathogenesis. We find several hypothetical proteins and assign them to virulence mechanisms, including attachment, cytoskeletal modulation, and autophagy evasion. The B. pseudomallei ‘TRANSITome’ provides prokaryotic single-cell transcriptomics information enabling high-resolution understanding of host-pathogen interactions., Prokaryotic cell transcriptomics has been limited to mixed or sub-population dynamics and individual cells within heterogeneous populations. Here the authors develop a ‘TRANSITomic’ approach to profile transcriptomes of single Burkholderia pseudomallei cells as they transit through host cell infection.

Fragmentation reactions that generate alkynes in conjunction with C–C bond cleavage require special considerations but can produce high-value alkyne building blocks for organic synthesis.

In the present work, self-healable rubber blends consisting of modified bromobutyl rubber (BIIR)/natural rubber (NR) blends filled with silica (Si)/carbon nanotube (CNT) hybrid filler were prepared as model system. It was found that an addition of a low loading of silica as secondary filler for CNTs significantly enhances the tensile strength of the BIIR/NR blends while maintaining the self-healing properties. The synergistic effect of the hybrid filler on the mechanical properties of BIIR/NR blends can be explained by taking into account the selective wetting of the hybrid filler. Thus, a new method based on the wetting concept has been further developed for quantification of the selective wetting of hybrid filler in the BIIR/NR blends. The migration of silica to the BIIR/NR interfacial region and preferential location of the CNTs in NR phase may impart the BIIR/NR blends better mechanical performance and high electrical conductivity. The BIIR/NR blends filled with hybrid filler Si/carbon black (CB) were prepared as reference system. Its similarities and differences with the model system were discussed regarding the selective filler wetting and localization.

Bacterial cooperative associations and dynamics in biofilm microenvironments are of special interest in recent years. Knowledge of localized gene-expression and corresponding bacterial behaviors within the biofilm architecture at a global scale has been limited, due to a lack of robust technology to study limited number of cells in stratified layers of biofilms. With our recent pioneering developments in single bacterial cell transcriptomic analysis technology, we generated herein an unprecedented spatial transcriptome map of the mature in vitro Pseudomonas aeruginosa biofilm model, revealing contemporaneous yet altered bacterial behaviors at different layers within the biofilm architecture (i.e., surface, middle and interior of the biofilm). Many genes encoding unknown functions were highly expressed at the biofilm-solid interphase, exposing a critical gap in the knowledge of their activities that may be unique to this interior niche. Several genes of unknown functions are critical for biofilm formation. The in vivo importance of these unknown proteins was validated in invertebrate (fruit fly) and vertebrate (mouse) models. We envisage the future value of this report to the community, in aiding the further pathophysiological understanding of P. aeruginosa biofilms. Our approach will open doors to the study of bacterial functional genomics of different species in numerous settings.

BACKGROUND: To compare functional staging classifications in Vietnamese patients with primary open angle glaucoma (POAG) and chronic primary angle closure glaucoma (PACG). METHODS: A retrospective cross-section study was conducted at a national setting. Two hundred seven eyes of 207 patients were recruited. Patients were tested with standard automated perimetry. Field loss was generally classified in four stages (normal, early, moderate, and severe), using four classification strategies: (1) Hodapp–Parrish–Anderson (HPA), (2) enhanced Glaucoma Staging System (eGSS), (3) modified Glaucoma Staging System (mGSS) and (4) the Advanced Glaucoma Intervention Study (AGIS). AGIS as a standard method was used to judge the staging performance of the other three classifications in terms of agreement (Cohen Kappa—K) and association (Chi-Square Test—Cramer’s V). RESULTS: The agreement between AGIS and mGSS (K = 0.687; p

Background: Clinically, there are many systems of visual field staging parallel exist. The agreement between glaucoma function and appearance is still uncertain. This study was to compare visual field staging classifications (as function staging) with disc damage likelihood scale (DDLS as appearance staging) in primary open angle glaucoma (POAG).Methods: Cross-section study on 55 patients (90 eyes) examined and treated in Vietnam National Institute of Ophthalmology from 9/2016 – 9/2017. The MD (mean deviation), PSD (pattern standard deviation), VFI (visual field index) indexes were recorded and were used to classify glaucoma staging according to categories of eGSS (enhance glaucoma severity staging), mGSS (modified glaucoma severity staging), HPA (Hoddaps – Palmela – Adersons), AGIS (advance glaucoma intervention study) and DDLS. Using SPSS 16.0 software, statistically significance was tested with proper tests as Chi – square test, Fisher exact test, Kappa (to assess the agreement - disagreement), Spearman Rank (to assess the level of correlation).Results: Mean visual field indexes were -11.49 dB (MD), 5.85 dB (PSD), 74.2% (VFI). Compared to the DDLS, all systems had slight agreement (K < 0.2). The disagreement percentage was highest in eGSS (70%) then followed by mGSS (50%) and HPA (48.9%). In terms of definitive diagnosis, HPA and mGSS showed a substantial agreement (K > 0.6) with AGIS which is higher than that between eGSS and AGIS (retrospectively 0.773 and 0.75 vs 0.399) with p < 0.001. In terms of staging detection, the agreement between HPA &mGSS and AGIS was substantial (K > 0.6) while that between eGSS and AGIS was fair (K < 0.4).Conclusion: mGSS and HPA tend to show the stronger agreement with standard classifications than eGSS. mGSS should be used in clinical practice and research.

Purpose: To identify risk factors for primary open-angle glaucoma (POAG) in a Vietnamese population. Methods: A matched case-control study was conducted at Vietnam National Institute of Ophthalmology. Cases were patients clinically diagnosed with POAG. Controls were caregivers of patients with other eye diseases, free from any ocular and systemic abnormalities, and were selected and then matched with the cases by gender and age. Cases and controls were examined by a glaucoma specialist to confirm presence or absence of POAG. Data on demographic, ophthalmic and medical conditions were collected via interview at time of examination. Associations between POAG and potential factors were examined by univariate conditional logistic regression analysis (OR – Odd Ratios) and multivariate conditional logistic regression (AOR – Adjusted Odd Ratios) with level of significance is 0.05. Results: In total, 128 cases with POAG and 128 controls were recruited. Multivariate conditional (fixed-effects) logistic regression analysis resulted in several significant risk factors, including hypertension AOR =4.7 (95% CI: 1.8–12.0; p=0.002); family history of glaucoma AOR = 6.4 (95% CI: 1.3 – 32.2; p = 0.026); over–the–counter eye medication AOR = 3.1 (95% CI: 1.5 – 6.5; p = 0.006), and a protective role for marriage AOR = 0.2 (95% CI: 0.1 – 0.7; p = 0.006) and weekly exercise AOR = 0.3 (95% CI: 0.1 – 0.8; p = 0.021). Conclusions: Hypertension, family history of glaucoma and over–the–counter eye medication were risk factors while marriage and weekly exercise were protective factors for POAG in Vietnamese population.

A total synthesis of the illudalane sesquiterpene illudinine was realized in eight steps and 14% overall yield from commercially available dimedone. The approach features tandem fragmentation/Knoevenagel-type condensation and microwave-assisted oxidative cycloisomerization to establish the isoquinoline core. Completion of the synthesis involves a recently reported cascade SNAr/Lossen rearrangement on a densely functionalized aryl bromide and an optimized procedure for O-methylation of 8-hydroxyisoquinolines. The oxidative cycloisomerization proceeds by way of a novel inverse-demand intramolecular dehydro-Diels–Alder cycloaddition, which has a potentially broader appeal for preparing substituted isoquinolines.

// Lin Zhou 1, * , Yuan Chang 1, * , Le Xu 2, * , Son Tung Nguyen Hoang 1 , Zheng Liu 3 , Qiang Fu 3 , Zongming Lin 1 , Jiejie Xu 3 1 Department of Urology, Zhongshan Hospital, Fudan University, Shanghai, China 2 Department of Urology, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, China 3 Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Shanghai, China * These authors have contributed equally to this work Correspondence to: Zongming Lin, email: zongminglin@gmail.com Jiejie Xu, email: jjxufdu@fudan.edu.cn Keywords: vascular mimicry, bladder cancer, recurrence, radical cystectomy, adjuvant chemotherapy Received: July 25, 2016 Accepted: October 04, 2016 Published: October 20, 2016 ABSTRACT Vascular mimicry (VM) refers to the plasticity of aggressive cancer cells forming de novo vascular networks, which promoted tumor metastasis. The aim of this study was evaluate the impact of VM on recurrence-free survival (RFS) in urothelial carcinoma of the bladder (UCB). Records from 202 patients treated with radical cystectomy (RC) for UCB at Zhongshan Hospital between 2002 and 2014 were reviewed. The presence of VM was identified by CD31-PAS double staining. Positive VM staining occurred in 19.3% (39 of 202) UCB cases, and it was associated with increased risks of recurrence (Log-Rank p

It is generally believed that the Pseudomonas aeruginosa biofilm matrix itself acts as a molecular sieve or sink that contributes to significant levels of drug resistance, but it is becoming more apparent that multidrug efflux pumps induced during biofilm growth significantly enhance resistance levels. We present here a novel transcriptional regulator, PA3898, which controls biofilm formation and multidrug efflux pumps in P. aeruginosa. A mutant of this regulator significantly reduced the ability of P. aeruginosa to produce biofilm in vitro and affected its in vivo fitness and pathogenesis in Drosophila melanogaster and BALB/c mouse lung infection models. Transcriptome analysis revealed that PA3898 modulates essential virulence genes/pathways, including multidrug efflux pumps and phenazine biosynthesis. Chromatin immunoprecipitation sequencing (ChIP-seq) identified its DNA binding sequences and confirmed that PA3898 directly interacts with promoter regions of four genes/operons, two of which are mexAB-oprM and phz2. Coimmunoprecipitation revealed a regulatory partner of PA3898 as PA2100, and both are required for binding to DNA in electrophoretic mobility shift assays. PA3898 and PA2100 were given the names MdrR1 and MdrR2, respectively, as novel repressors of the mexAB-oprM multidrug efflux operon and activators for another multidrug efflux pump, EmrAB. The interaction between MdrR1 and MdrR2 at the promoter regions of their regulons was further characterized via localized surface plasmon resonance and DNA footprinting. These regulators directly repress the mexAB-oprM operon, independent of its well-established MexR regulator. Mutants of mdrR1 and mdrR2 caused increased resistance to multiple antibiotics in P. aeruginosa, validating the significance of these newly discovered regulators.

Natural competency requires uptake of exogenous DNA from the environment and the integration of that DNA into recipient bacteria can be used for DNA-repair or genetic diversification. The Burkholderia genus is unique in that only some of the species and strains are naturally competent. We identified and characterized two genes, comE and crp, from naturally competent B. pseudomallei 1026b that play a role in DNA uptake and catabolism. Single-copies of rhamnose-inducible comE and crp genes were integrated into a Tn7 attachment-site in non-naturally competent Burkholderia including pathogens B. pseudomallei K96243, B. cenocepacia K56-2, and B. mallei ATCC23344. Strains expressing comE or crp were assayed for their ability to uptake and catabolize DNA. ComE and Crp allowed non-naturally competent Burkholderia species to catabolize DNA, uptake exogenous gfp DNA and express GFP. Furthermore, we used synthetic comE and crp to expand the utility of the λ-red recombineering system for genetic manipulation of non-competent Burkholderia species. A newly constructed vector, pKaKa4, was used to mutate the aspartate semialdehyde dehydrogenase (asd) gene in four B. mallei strains, leading to the complete attenuation of these tier-1 select-agents. These strains have been excluded from select-agent regulations and will be of great interest to the field.

3,3-Dimethylcyclopentanes (neopentylenes) are ubiquitous in Nature but largely absent from synthetic pharmaceutical libraries. Neopentylenes define a hydrophobic and rigid 3-D topology with distinct molecular pharmacology, as exemplified here with two neopentylene-fused analogues of the synthetic anti-inflammatory drug, ibuprofen.

Customized architecture is widely recognized as an important approach for improved performance and energyefficiency. To balance generality and customization benefit, researchers have proposed to federate heterogeneous micro-engines. Using the 10x10 architecture and an integrated image and vision benchmark as a case study, we explore the performance and energy benefits achievable. Results for current 32nm technology and DDR3 memory show 10x10 architecture benefits of 140x performance and 72x energy overall. Adding 3D-stacked DRAM increase benefits to 171x (performance) and 100x (energy). Finally, considering future 7nm transistor process, benefits as large as 597x (performance) and 137x energy are observed.

Until recently, transcriptome analyses of single cells have been confined to eukaryotes. The information obtained from single-cell transcripts can provide detailed insight into spatiotemporal gene expression, and it could be even more valuable if expanded to prokaryotic cells. Transcriptome analysis of single prokaryotic cells is a recently developed and powerful tool. Here we describe a procedure that allows amplification of the total transcript of a single prokaryotic cell for in-depth analysis. This is performed by using a laser-capture microdissection instrument for single-cell isolation, followed by reverse transcription via Moloney murine leukemia virus, degradation of chromosomal DNA with McrBC and DpnI restriction enzymes, single-stranded cDNA (ss-cDNA) ligation using T4 polynucleotide kinase and CircLigase, and polymerization of ss-cDNA to double-stranded cDNA (ds-cDNA) by Φ29 polymerase. This procedure takes ∼5 d, and sufficient amounts of ds-cDNA can be obtained from single-cell RNA template for further microarray analysis.

Burkholderia spp. are genetically and physiologically diverse. Some strains are naturally transformable and capable of DNA catabolism. Burkholderia pseudomallei (Bp) strains 1026b and K96243 and B. thailandensis strain E264 are able to utilize DNA as a sole carbon source for growth, while only strains 1026b and E264 are naturally transformable. In this study, we constructed low-copy broad-host-range fosmid library, containing Bp strain 1026b chromosomal DNA fragments, and employed a novel positive selection approach to identify genes responsible for DNA uptake and DNA catabolism. The library was transferred to non-competent Bp K96243 and B. cenocepacia (Bc) K56-2, harboring chromosomally-inserted FRT-flanked sacB and pheS counter-selection markers. The library was incubated with DNA encoding Flp recombinase, followed by counter-selection on sucrose and chlorinated phenylalanine, to select for clones that took up flp-DNA, transiently expressed Flp, and excised the sacB-pheS cassette. Putative clones that survived the counter-selection were subsequently incubated with gfp-DNA and bacteria were visualized via fluorescent microscopy to confirm natural competency. Fosmid sequencing identified several 1026b genes implicated in DNA uptake, which were validated using chromosomal mutants. One of the naturally competent clones selected in Bc K56-2 enabled Bc, Bp and B. mallei to utilize DNA as a sole carbon source, and all fosmids were used to successfully create mutations in non-naturally-competent B. mallei and Bp strains.

A rhodium-catalyzed cycloisomerization and oxidation of tethered dienynes for the synthesis of indanes is described. An auxiliary fragmentation/olefination method (also described herein) provides novel access to tethered alkyne-dienoate substrates. The reported method circumvents current limitations in and expands the scope of inverse-demand Diels–Alder-type cycloadditions. Traditional discovery substrates involving malonate-, ether-, and sulfonamide-based tethers are problematic in the current methodology, underscoring the unique virtue of neopentylene-tethered substrates for reaction discovery.

Sand production in Sandstone reservoirs is a complex problem to Oil & Gas companies. Many methods have used to solve this problem but these methods only have effect for the first period of production without effective in long time. Sand production causes big damages such as: producing tools corrosion in hole, separating tools corrosion at surface, formation collapse, so sand production in well is always the urgent problem. Based on formation characteristic research, advantages and disadvantages of sand failure analysis and combining the advanced methods, this study introduces Production Sand Pressure Model to predict sand productivity in sedimentary reservoirs in field X in Cuu Long basin.

Strategic pairing of ring openings and cycloisomerization provides rapid and efficient “open and shut” entry into sparsely functionalized illudalanes, as exemplified here in the context of a six-step synthesis of alcyopterosin A. Key steps include a tandem ring-opening fragmentation/olefination process for preparing a neopentyl-tethered 1,6-enyne, ring-opening olefination telescoped with alkyne homologation, and Rh-catalyzed oxidative cycloisomerization.

The performance of a brackish water reverse osmosis membrane in the presence of a model polysaccharide foulant, sodium alginate, is investigated. The concentration of foulant, the feed pH and the presence of calcium are all shown to impact upon this performance. The flux decline is greatest at low feed pH, high calcium concentrations and high foulant concentrations. Sodium and chloride rejection falls as foulant concentration increases, due to cake-enhanced osmotic pressure. Effects are again greatest at low feed pH and high calcium concentrations. In part, this reflects a shift of the isoelectric point of the membrane from 4.3 to below 3.0 as alginate is added, reflecting the lower pK a of the alginate foulant. Conversely, calcium rejection improves in the presence of even small quantities of alginate foulant at all pH values. These increases in rejection are believed to relate to a direct interaction between the dissolved ions and the alginate which act to sequester the salt.

Phage λ-Red proteins are powerful tools for pulling and knocking out chromosomal fragments but have been limited to the γ-proteobacteria. Procedures are described here to easily knock out (KO) and pull out (PO) chromosomal DNA fragments from naturally transformable Burkholderia thailandensis and Burkholderia pseudomallei. This system takes advantage of published compliant counterselectable and selectable markers (sacB, pheS, gat and the arabinose-utilization operon) and λ-Red mutant proteins. pheS-gat (KO) or oriT-ColE1ori-gat-ori1600-rep (PO) PCR fragments are generated with flanking 40- to 45-bp homologies to targeted regions incorporated on PCR primers. One-step recombination is achieved by incubation of the PCR product with cells expressing λ-Red proteins and subsequent selection on glyphosate-containing medium. This procedure takes ~10 d and is advantageous over previously published protocols: (i) smaller PCR products reduce primer numbers and amplification steps, (ii) PO fragments suitable for downstream manipulation in Escherichia coli are obtained and (iii) chromosomal KO increases flexibility for downstream processing.