Sec22b and Stx4 Depletion Has No Major Effect on Cross-Presentation of PLGA Microsphere-Encapsulated Antigen and a Synthetic Long Peptide In Vitro.

The induction of CTL responses by vaccines is important to combat infectious diseases and cancer. Biodegradable poly(lactic-co-glycolic acid) (PLGA) microspheres and synthetic long peptides are efficiently internalized by professional APCs and prime CTL responses after cross-presentation of Ags on MHC class I molecules. Specifically, they mainly use the cytosolic pathway of cross-presentation that requires endosomal escape, proteasomal processing, and subsequent MHC class I loading of Ags in the endoplasmic reticulum (ER) and/or the endosome. The vesicle SNARE protein Sec22b has been described as important for this pathway by mediating vesical trafficking for the delivery of ER-derived proteins to the endosome. As this function has also been challenged, we investigated the role of Sec22b in cross-presentation of the PLGA microsphere-encapsulated model Ag OVA and a related synthetic long peptide. Using CRISPR/Cas9-mediated genome editing, we generated Sec22b knockouts in two murine C57BL/6-derived APC lines and found no evidence for an essential role of Sec22b. Although pending experimental evidence, the target SNARE protein syntaxin 4 (Stx4) has been suggested to promote cross-presentation by interacting with Sec22b for the fusion of ER-derived vesicles with the endosome. In the current study, we show that, similar to Sec22b, Stx4 knockout in murine APCs had very limited effects on cross-presentation under the conditions tested. This study contributes to characterizing cross-presentation of two promising Ag delivery systems and adds to the discussion about the role of Sec22b/Stx4 in related pathways. Our data point toward SNARE protein redundancy in the cytosolic pathway of cross-presentation.

Crystalline Porous Organic Cage Membranes Constructed Using Fortified Intermolecular Interactions for Molecular Sieving.

Among the various types of materials with intrinsic porosity, porous organic cages (POCs) are distinctive as discrete molecules that possess intrinsic cavities and extrinsic channels capable of facilitating molecular sieving. However, the fabrication of POC membranes remains highly challenging due to the weak noncovalent intermolecular interactions and most reported POCs are powders. In this study, we constructed crystalline free-standing porous organic cage membranes by fortifying intermolecular interactions through the induction of intramolecular hydrogen bonds, which was confirmed by single-crystal X-ray analysis. To elucidate the driving forces behind, a series of terephthaldehyde building blocks containing different substitutions were reacted with flexible triamine under different conditions via interfacial polymerization (IP). Furthermore, density functional theory (DFT) calculations suggest that intramolecular hydrogen bonding can significantly boost the intermolecular interactions. The resulting membranes exhibited fast solvent permeance and high rejection of dyes not only in water, but also in organic solvents. In addition, the membrane demonstrated excellent performance in precise molecular sieving in organic solvents. This work opens an avenue to designing and fabricating free-standing membranes composed of porous organic materials for efficient molecular sieving.

Prevalence and type-specific distribution of human papillomavirus infection among women in mid-western rural, Nepal- A population-based study.

BACKGROUND: Cervical cancer is the most common cancer among women in Nepal. The prevalence of human papillomavirus (HPV) 16 and or HPV 18 among women with cervical pre-cancer and cancer is higher than the incidence of HPV in the world population. The population-based epidemiological data of HPV in the general population in most parts of the country remains unknown. The objective of this study was to assess the prevalence and type distribution of HPV infection and association of abnormal cytology with high risk HPV infection among women in mid-western rural, Nepal. METHODS: A population-based cross sectional study was conducted in Jumla, one of the most remote districts in Nepal. A total of 1050 cervical samples were collected from married and non- pregnant women aged 20-65 years during mobile Cervical Cancer Screening Clinics conducted from May 2016 to January 2017. The presence of HPV DNA was firstly confirmed by HPV consensus PCR using PGMY09/PGMY11 designed primers, then HPV positive samples were further genotyped by the membrane hybridization method to detect the 21 high-risk HPV (HR-HPV) and low-risk HPV types. The prevalence of HR-HPV among women with normal and abnormal cytology was calculated. Data were analyzed using SPSS software for Windows. P < 0.05 was considered statistically significant. RESULTS: A total of 998 women were eligible for this study with the mean age 32.6 ± 8.6 years, and the mean marital age was 16.7 ± 3.8 years. The overall prevalence of HPV infections was 19.7%. HR-HPV and low-risk HPV were 11.7 and 8.7% respectively. The six most common HR-HPV types were HPV16, 39, 58, 33, 51 and 18. HR-HPV infection among the women with abnormal and normal cytology was of 27.3 and 10.8% respectively. CONCLUSIONS: There was a higher prevalence of HR-HPV infection among women living in Jumla than other parts of Nepal. This study provides preliminary information on overall HPV and type-specific HR-HPV prevalence, HR-HPV 16, 39, 58, 33, 51, and 18 are the most prevalent genotypes in this region. The data contribute to the epidemiological knowledge about HPV and type-specific HR-HPV genotypes prevalence in mid-Western Nepal.

Covalent Organic Network Membranes with Tunable Nanoarchitectonics from Macrocycle Building Blocks for Graded Molecular Sieving.

Traditional piperazine-based polyamide membranes usually suffer from the intrinsic trade-off relationship between selectivity and permeance. The development of macrocycle membranes with customized nanoscale pores is expected to address this challenge. Herein, we introduce 1,4-diazacyclohexane (2N), 1,4,7-triazacyclononane (3N), and 1,4,8,11-tetraazacyclotetradecane (4N) as molecular building blocks to construct the nanoarchitectonics of polyamide membranes prepared from interfacial polymerization (IP). The permeance of covalent organic network membranes follows the trend of 4N-TMC > 3N-TMC > 2N-TMC, while the molecular weight cutoff (MWCO) also follows the same trend of 4N-TMC > 3N-TMC > 2N-TMC, according to their nanopore size of the membranes. The microporosity, orientation, and surface chemistry of covalent organic network membranes can be rationally designed by macrocycle building units. The ordered nanoarchitectonics allows the membranes to attain an excellent performance in graded molecular sieving. Importantly, the novel covalent organic network membranes with tunable nanoarchitectonics prepared from macrocycle building units exhibited high water permeance (32.5 LMH/bar) and retained long-term stability after 100 h of test and bovine serum albumin fouling. These results reveal the enormous potential of 3N-TMC and 4N-TMC membranes in saline textile wastewater treatments and precise molecular sieving.

Lack of T04C9.1, the Homologue of Mammalian APPL2, Leads to Premature Ageing and Shortens Lifespan in Caenorhabditis elegans.

Ageing has been identified as an independent risk factor for various diseases; however, the physiological basis and molecular changes related to ageing are still largely unknown. Here, we show that the level of APPL2, an adaptor protein, is significantly reduced in the major organs of aged mice. Knocking down APPL2 causes premature ageing of human umbilical vein endothelial cells (HUVECs). We find that a lack of T04C9.1, the homologue of mammalian APPL2, leads to premature ageing, slow movements, lipid deposition, decreased resistance to stresses, and shortened lifespan in Caenorhabditis elegans (C. elegans), which are associated with decreased autophagy. Activating autophagy by rapamycin or inhibition of let-363 suppresses the age-related alternations, impaired motility, and shortened lifespan of C. elegans, which are reversed by knocking down autophagy-related genes. Our work provides evidence that APPL2 and its C. elegans homologue T04C9.1 decrease with age and reveals that a lack of T04C9.1 bridges autophagy decline and ageing in C. elegans.

Ultrafast Interfacial Self-Assembly toward Bioderived Polyester COF Membranes with Microstructure Optimization.

The precise manipulation of the microstructure (pore size, free volume distribution, and connectivity of the free-volume elements), thickness, and mechanical characteristics of membranes holds paramount significance in facilitating the effective utilization of self-standing membranes. In this contribution, the synthesis of two innovative ester-linked covalent-organic framework (COF) membranes is first reported, which are generated through the selection of plant-derived ellagic acid and quercetin phenolic monomers in conjunction with terephthaloyl chloride as a building block. The optimization of the microstructure of these two COF membranes is systematically achieved through the application of three different interfacial electric field systems: electric neutrality, positive electricity, and negative electricity. It is observed that the positively charged system facilitates a record increase in the rate of membrane formation, resulting in a denser membrane with a uniform pore size and enhanced flexibility. In addition, a correlation is identified wherein an increase in the alkyl chain length of the surfactants leads to a more uniform pore size and a decrease in the molecular weight cutoff of the COF membrane. The resulting COF membrane exhibits an unprecedented combination of high water permeance, superior sieving capability, robust mechanical strength, chemical robustness for promising membrane-based separation science and technology.

Microporous polyarylate membranes based on 3D phenolphthalein for molecular sieving.

Thin-film composite (TFC) membranes have gradually replaced some traditional technologies in the extraction, separation, and concentration of high value-added pharmaceutical ingredients due to their controllable microstructure. Nevertheless, devising solvent-stable, scalable TFC membranes with high permeance and efficient molecule selectivity is urgently needed to improve the separation efficiency in the separation process. Here, we propose phenolphthalein, a commercial acid-base indicator, as an economical monomer for optimizing the micropore structure of selective layers with thickness down to 30 nanometers formed by in situ interfacial reactions. Molecular dynamics simulations indicate that the polyarylate membranes prepared using three-dimensional phenolphthalein monomers exhibit tunable microporosity and higher pore interconnectivity. Moreover, the TFC membranes show a high methanol permeance (9.9 ± 0.1 liters per square meter per hour per bar) and small molecular weight cutoff (≈289 daltons) for organic micropollutants in organic solvent systems. The polyarylate membranes exhibit higher mechanical strength (2.4 versus 0.8 gigapascals) compared to the traditional polyamide membrane.

Flexible Covalent Organic Network with Ordered Honeycomb Nanoarchitecture for Molecular Separations.

Membranes with precisely defined nanostructure are desirable for energy-efficient molecular separations. The emergence of membranes with honeycomb lattice or topological nanopores is of fundamental importance. The tailor-made nanostructure and morphology may have huge potential to resolve the longstanding bottlenecks in membrane science and technology. Herein, inspired by honeycomb architecture, we demonstrate an effective and scalable route based on interfacial polymerization (IP) to generate flexible and ordered covalent organic network (CON) membranes for liquid-phase molecular separations. The aperture size of a CON membrane can be reasonably designed through the strong covalent bond between molecular building blocks. The fabricated CON membrane formed by IP showed an obviously size-dependent sieving of molecules, yielding a stepwise conversion from low rejection to the expected high rejection. Moreover, the CON membrane was also found to have the sieving capability for tetracycline and ciprofloxacin, ascribed to the effect of size exclusion by an ordered single-nanoscale channel (<1 nm). This approach provides a viable strategy for creating target-sized channels from molecular-level design and demonstrates their potential for accurate molecular separations.

Nanoarchitectonics of carbon molecular sieve membranes with graphene oxide and polyimide for hydrogen purification.

Hydrogen is an important energy carrier for the transition to a carbon-neutral society, the efficient separation and purification of hydrogen from gaseous mixtures is a critical step for the implementation of a hydrogen economy. In this work, graphene oxide (GO) tuned polyimide carbon molecular sieve (CMS) membranes were prepared by carbonization, which show an attractive combination of high permeability, selectivity and stability. The gas sorption isotherms indicate that the gas sorption capability increases with the carbonization temperature and follows the order of PI-GO-1.0%-600 °C > PI-GO-1.0%-550 °C > PI-GO-1.0%-500 °C, more micropores would be created under higher temperatures under GO guidance. The synergistic GO guidance and subsequent carbonization of PI-GO-1.0% at 550 °C increased H2 permeability from 958 to 7462 Barrer and H2/N2 selectivity from 14 to 117, superior to state-of-the-art polymeric materials and surpassing Robeson's upper bound line. As the carbonization temperature increased, the CMS membranes gradually changed from the turbostratic polymeric structure to a denser and more ordered graphite structure. Therefore, ultrahigh selectivities for H2/CO2 (17), H2/N2 (157), and H2/CH4 (243) gas pairs were achieved while maintaining moderate H2 gas permeabilities. This research opens up new avenues for GO tuned CMS membranes with desirable molecular sieving ability for hydrogen purification.

A 2D Soft Covalent Organic Framework Membrane Prepared via a Molecular Bridge.

Highly flexible and robust self-standing covalent organic framework (COF) membranes with rapid preparation are important but technically challenging for achieving precise separation. Herein , a novel imine-based 2D soft covalent organic framework (SCOF) membrane with a large area of 226.9 cm2 , via ingeniously selecting an aldehyde flexible linker and a trigonal building block, is reported. The soft 2D covalent organic framework membrane is rapidly formed (≈5 min) based on the sodium dodecyl sulfate (SDS) molecular channel constructed at the water/dichloromethane (DCM) interface, which is the record-fast SCOF membrane formation and 72 times faster than that in the reported literature. MD simulation and DFT calculation elucidate that the dynamic, self-assembled SDS molecular channel facilitates faster and more homogeneous transfer of amine monomers in the bulk, thereby forming a soft 2D self-standing COF membrane with more uniform pores. The formed SCOF membrane exhibits superb sieving capability for small molecules, robustness in strong alkaline (5 mol L-1 NaOH), acid (0.1 mol L-1 HCl), and various organic solutions, and sufficient flexibility with a large curvature of 2000 m-1 for membrane-based separation science and technology.

Prognostic Significance of Log(CA125)/PCI for the Resectability of Epithelial Ovarian Cancer: A Retrospective Study.

OBJECTIVE: This study aimed to evaluate the roles of the ratio of log(serum CA125 level)/PCI in epithelial ovarian cancer. METHODS: This is a retrospective study. Data were retrieved for patients with epithelial ovarian cancer who received primary debulking surgeries (PDS) between January 2014 and December 2017 in Zhongnan Hospital of Wuhan University. The PCI and CA125 were determined retrospectively using surgical reports, histological findings, and intraoperative photographic documentation. Survival analysis and ROC curves were applied to evaluate the roles of the ratio of log(serum CA125 level)/PCI in epithelial ovarian cancer. RESULTS: A total of 69 patients were included. Of these, serous ovarian cancer and mucinous carcinoma accounted for 63.8% (n=44) and 31.9% (n=22), respectively. The remaining patients had clear cell carcinoma (2.9%, n=2) and endometrioid carcinoma ( 1.4%, n= 1). Kaplan-Meier survival analysis showed that log(serum CA125 level)/PCI (log-rank p=0.018) were prognostic factors for OS. Cox regression analysis, otherwise, suggested that only stages were an independent factor of PFS (P=0.02, 95% CI 0.043-0.763); outcomes of cytoreductive surgery could only affect OS significantly (P=0.009, 95% CI 1.639-31.016). Binary logistic regression discovered that only log(serum CA125 level)/PCI was an independent risk factor of PDS. We further used the ROC curve to find that log(serum CA125 level)/PCI could correctly predict the resectability of PDS with AUC 0.781. CONCLUSION: The ratio of log(CA125)/PCI that combined the tumor burden and characteristics of peritoneal carcinoma of ovarian origin can predict the resectability of PDS in epithelial ovarian cancer.

The effect of HMGB1 on the clinicopathological and prognostic features of cervical cancer.

Cervical cancer is the third leading cause of cancer death among women in less-developed regions. Because of the poor survivorship of patients with advanced disease, finding new biomarkers for prognostic prediction is of great importance. In the current study, mRNA datasets (GSE9750 and GSE63514) were retrieved from Gene Expression Omnibus and was used to identify differentially expressed genes. The underlying molecular mechanisms associated with high-mobility group box 1 protein (HMGB1) were investigated using bioinformatics analysis. Immunohistochemical analysis of HMGB1 was performed on 239 cases of cervical cancer samples to investigate its possible correlation with clinicopathological characteristics and outcomes. A preliminary validation has been made to explore the possible correlation factors with HMGB1 that promote migration of cervical cancer cells. Bioinformatics analysis showed that adherens junction was significant for both P-value and enrichment scores, which was consistent with the clinical study. The underlying molecular mechanisms might be the interaction among HMGB1, RAC1, and CDC42. HMGB1 expression was significantly associated with tumor size, parametrial infiltration, the depth of cervical stromal invasion, and FIGO stage (P=0.003, 0.019, 0.013, and 0.003, respectively). FIGO stage, lymph mode metastasis, and HMGB1 expression were independent predictors of a poorer prognosis of patients with cervical cancer. Knockdown of HMGB1 inhibits migration of Siha and C33A cells in vitro Western blot and quantitative real-time PCR (qRT-PCR) showed that the expression of RAC1 and CDC42 was positively correlated with HMGB1. HMGB1 is a useful prognostic indicator and a potential biomarker of cervical cancer. RAC1 and CDC42 may be involved in the progression of cervical cancer migration induced by HMGB1.

Ultrasound-mediated interfacial protein adsorption and fat crystallization in cholesterol-reduced lard emulsion.

Lard with a substantially reduced cholesterol content through aqueous enzyme extraction is an attractive source of lipid in healthy and nutritious emulsion food product development. The objective of this study was to elucidate the crystallization behavior (4-20 °C) of emulsions prepared from low-cholesterol lard in relation to protein emulsifier (2 and 4% whey protein isolate, WPI) and ultrasound (475 w, 5 min) treatments. The physicochemical properties and fat crystallization pattern of the emulsions were investigated. Emulsions with 4% WPI were superior to those with 2% WPI on interfacial adsorption and crystal size reduction. Ultrasonic treatment of prepared emulsions further decreased fat crystal size, promoted small and uniform crystal distribution, and slightly destabilized emulsions. Protein concentration and ultrasonic treatment had no obvious effect on crystal transition from ß' to the more stable ß-form based on X-ray diffraction. The melting properties determined by differential scanning calorimetry indicated that emulsified lard had a lower onset melting temperature than bulk lard, and with ultrasound treatment, the melting enthalpy increased remarkably. The ultrasound-induced change in fat crystalline structure and emulsion meta-stability may be valuable to the manufacture of healthy, emulsion-incorporated food products.

Peripheral blood T cell dynamics predict relapse in multiple sclerosis patients on fingolimod.

BACKGROUND: Fingolimod efficiently reduces multiple sclerosis (MS) relapse by inhibiting lymphocyte egress from lymph nodes through down-modulation of sphingosine 1-phosphate (S1P) receptors. We aimed to clarify the alterations in peripheral blood T cell subsets associated with MS relapse on fingolimod. METHODS/PRINCIPAL FINDINGS: Blood samples successively collected from 23 relapsing-remitting MS patients before and during fingolimod therapy (0.5 mg/day) for 12 months and 18 healthy controls (HCs) were analysed for T cell subsets by flow cytometry. In MS patients, the percentages of central memory T (CCR7+CD45RO+) cells (TCM) and naïve T (CCR7+CD45RO-) cells decreased significantly, while those of effector memory T (CCR7-CD45RA-) and suppressor precursor T (CD28-) cells increased in both CD4+T and CD8+T cells from 2 weeks to 12 months during fingolimod therapy. The percentages of regulatory T (CD4+CD25highCD127low) cells in CD4+T cells and CCR7-CD45RA+T cells in CD8+T cells also increased significantly. Eight relapsed patients demonstrated greater percentages of CD4+TCM than 15 non-relapsed patients at 3 and 6 months (p=0.0051 and p=0.0088, respectively). The IL17-, IL9-, and IL4-producing CD4+T cell percentages were significantly higher at pre-treatment in MS patients compared with HCs (p<0.01 for all), while the IL17-producing CD4+T cell percentages tended to show a transient increase at 2 weeks of fingolimod therapy (pcorr=0.0834). CONCLUSIONS: The CD4+TCM percentages at 2 weeks to 12 months during fingolimod therapy are related to relapse.