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Accumulating evidence has demonstrated that neural stem cells (NSCs) have regenerative capacity after brain injuries, such as in aneurysmal subarachnoid hemorrhage (SAH). The reactive oxygen species (ROS)-induced NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome triggers inflammatory responses and pyroptosis of cells; however, whether ROS-induced neuroinflammation modulates the fate of endogenous NSCs after SAH remains largely unknown. In this study, the level of IL-1β was increased in the cerebrospinal fluid (CSF) of patients with SAH. In an endovascular perforation model of SAH in mice, the secretion of IL-1β increased to a peak at 24 h following SAH, and the expression of Caspase1 and NLRP3 was elevated in the hippocampus. Primary cultured NSCs were incubated with hemoglobin (Hb) to mimic SAH in vitro. The cell viability, LDH release, intracellular ROS levels, scanning electron microscopy (SEM), and the expression of NLRP3 and pyroptosis indicators (GSDMD, ASC, and Caspase-1) in NSCs after SAH were examined to investigate the process of pyroptosis. We found that pyroptotic death featuring cellular swelling, cell membrane pore formation and elevated IL-1β was increased in cultured primary NSCs after Hb treatment, as was the expression of NLRP3, ASC, Caspase-1, and GSDMD. In addition, we found that ROS-induced pyroptosis of NSCs by activating the NLRP3/GSDMD pathway. These findings suggest that pyroptosis of NSCs induced by Hb can impede neural regeneration after SAH.

The electrocatalytic CO2 reduction reaction (CO2RR) decreases the amount of greenhouse gas in the atmosphere while enabling a closed carbon cycle. Herein, iron oleate was used as a precursor to produce oleic acid-coated triiron tetraoxide nanoparticles (Fe3O4@OA NPs) by pyrolysis, which was then assembled with reduced graphene oxide (rGO) and doped with dicyandiamide as a nitrogen source to obtain nitrogen-doped iron carbide nanoparticles assembled on rGO (N-Fe3C/rGO NPs). The catalyst prepared by nitrogen doping at 800 °C with an Fe3O4@OA NPs to rGO weight ratio of 20:1 showed good activity and stability for the CO2RR. At −0.3 to −0.4 V, the H2/CO ratio of the product from the catalyzed CO2RR was close to 2; thus, the product can be used for Fischer-Tropsch synthesis. The results of a series of experiments and X-ray photoelectron spectroscopy analysis showed that the synergy between the C N and Fe N groups in the catalyst can promote the reduction of CO2 to CO. This work demonstrates a facile method for improving the catalytic reduction of CO2.

Accumulating evidence suggests that SARS-CoV-2 impairs the adaptive immune system during acute infection. Still, it remains largely unclear whether the frequency and functions of T and B cells return to normal after the recovery of COVID-19. Here, we analyzed immune repertoires and SARS-CoV-2-specific neutralization antibodies in a prospective cohort of 40 COVID-19 survivors with a six-month follow-up after hospital discharge. Immune repertoire sequencing revealed abnormal T- and B-cell expression and function with large TCR/BCR clones, decreased diversity, abnormal class switch recombination and somatic hypermutation. A decreased number of B cells but an increased proportion of CD19

1q gain (+1q) is the most common high-risk cytogenetic abnormality (HRCA) in patients with multiple myeloma (MM). However, its prognostic value remains unclear in the era of novel agents. Here, we retrospectively analyzed the impact of +1q on the outcomes of 934 patients newly diagnosed with MM. +1q was identified in 53.1% of patients and verified as an independent variate for inferior overall survival (OS) (hazard ratio, 1.400; 95% confidence interval, 1.097-1.787; p = .007). Concurrence of other HRCAs (particularly t(14;16) and del(17p)) further exacerbated the outcomes of patients with +1q, suggesting prognostic heterogeneity. Thus, a risk-scoring algorithm based on four risk variates (t(14;16), hypercalcemia, ISS III, and high LDH) was developed to estimate the outcomes of patients with +1q. Of the patients, 376 evaluable patients with +1q were re-stratified into low (31.6%), intermediate (61.7%), and high risk (6.7%) groups, with significantly different progression-free survival and OS (p .0001), in association with early relapse of the disease. The prognostic value of this model was validated in the CoMMpass cohort. While attaining undetectable MRD largely circumvented the adverse impact of +1q, it scarcely ameliorated the outcome of the patients with high risk, who likely represent a subset of patients with extremely poor survival. Hence, patients with +1q are a heterogeneous group of high-risk patients, therefore underlining the necessity for their re-stratification. The proposed simple risk-scoring model can estimate the outcomes of patients with +1q, which may help guide risk-adapted treatment for such patients.

Aims: In this study, the dosage regimen establishment of cyadox nanosuspension against dairy cow mastitis caused by Staphylococcus aureus (S. aureus) was used as example to provide a general reference for the other novel nanocrystal preparations. Methods: The effect of cyadox against S. aureus isolates from dairy cows were firstly estimated and then the dosing regimen of nanosuspension after intramammary administration was optimized according to the model of ex vivo pharmacokinetic (PK) and pharmacodynamic (PD). The therapeutic efficacy of the predicted dosage regimen was evaluated. Results: The results demonstrated that cyadox has a concentration-dependent effect on S. aureus. The smallest and highest values of minimum inhibitory concentration (MIC) against 80 isolates was 8 and 64 μg/mL, respectively. The corresponding MIC 50 and MIC 90 was 16 and 32 μg/mL, respectively. The MIC against the pathogenic S. aureus SAHZ156001 in broth and milk were 16 and 32 μg/mL, respectively. The AUC 0-last and C max of cyadox in milk were 4442.877 μg*h/mL and 753.052 μg/mL, respectively. According to the inhibitory sigmoid E max modeling and dosage equation, the daily doses were predicted 1.6, 6.6, and 12.2 mL/gland to achieve bacteriostatic, bactericidal, and elimination effects. The dosage internal was daily administration for continuous three days. Conclusion: The clinical experiment showed that the efficient rates were 100, 100, and 90.9%, and the curative rates were 100, 81.8, and 63.6% in 12.2, 6.6 and 1.6 ml/gland groups, respectively. These results showed that cyadox nanosuspension had a good prospect as intramammary infusion to cure dairy cow mastitis infected by S. aureus. This study will be helpful for providing reference for nanocrystal preparation dosage regimen formulation.

An unparalleled [4 + 2]-annulation of prop-2-ynylsulfonium salts with isatoic anhydrides was developed, affording a series of 4-quinolones with a alkylthio group in medium to good yields under mild conditions. In this reaction type, the prop-2-ynylsulfonium salt serves as a C2 synthon and sulfide does not act as a leaving group, providing facile access to organosulfur compounds. The resulting quinolone products could be further transformed to a diverse range of synthetically useful compounds.

Arsenopyrite (FeAsS) and realgar (As4S4) are two common arsenic minerals that often cause serious environmental issues. Centralised treatment of arsenic-containing tailings can reduce land occupation and save management costs. The current work examined the remediation schemes of tailings from Hunan Province, China, where by different tailings containing arsenopyrite and realgar were blended with exogenous slag zero valence iron (ZVI). Introducing Fe-oxidising bacteria (Acidithiobacillus ferrooxidans) recreates a biologically oxidative environment. All bioleaching experiments were done over three stages, each for 7 days and the solid phase of all tests was characterised by scanning electron microscopy, X-ray diffraction, Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy and selective extraction analyses. The results showed that the mixture group reduced arsenic release by 72.9–74.7% compared with the control group. The addition of 0.2 g ZVI clearly decreased arsenic release, and the addition of 4.0 g ZVI led to the lowest arsenic release among all tests. The decrease of arsenic released from the tailings was due to the adsorption and uptake of arsenic by secondary iron-containing minerals and Fe–As(V) secondary mineralisation. The addition of large amounts of ZVI reduced the arsenic detected in the amorphous Fe precipitates. Therefore, a low cost and integrated strategy to reduce arsenic release from tailings is to mix two typical tailings and apply exogenous slag ZVI, which can apply to thein situremediation of two kinds or more arsenic-containing tailings.

Graphical abstract, Since the 1950s, great efforts have been made to develop antiviral agents against many infectious diseases such as human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV), human cytomegalovirus (HCMV), herpes simplex virus (HSV), and varicella-zoster virus (VZV). Among the list of nearly 106 antiviral agents approved in the past five decades, Prof. Erik De Clercq has contributed to the development of 7 antiviral drugs: tenofovir disoproxil fumarate (Viread®) for HIV and HBV treatment, tenofovir alafenamide (Vemlidy®) for HIV and HBV treatment, brivudine (Zostex®) for HSV-1 and VZV treatment, valacyclovir (Valtrex®) for HSV and VZV treatment, adefovir dipivoxil (Hepsera®) for HBV treatment, stavudine (Zerit®) for HIV treatment, and cidofovir (Vistide®) for treating HCMV retinitis in AIDS patients. In addition to the above antiviral drugs, his contributions include two anti-cancer drugs: rabacfosadine (Tanovea®-CA1) for canine lymphoma and plerixafor (Mozobil®) for multiple myeloma and non-Hodgkin’s lymphoma. These achievements are driven by his life-long passions for antiviral research and successful collaborations worldwide. To honor the 80th birthday of Prof. Erik De Clercq, this study highlights his scientific achievements and the importance of life-long passions and collaborations in the success of antiviral research and drug development.

With the development of frame materials, metal organic frameworks (MOFs) have been successfully applied in the fields of biological small molecule analysis and fluorescent DNA detection. In this work, in view of the good adsorption characteristics of MIL-101(Cr), the highly sensitive detection of adenosine triphosphate (ATP) assisted nucleic acid exonuclease amplification by MIL-101(Cr) on the different affinity of single stranded DNA and double stranded DNA was investigated. The detection limit of ATP reaches 1.7 μM, and the platform has good applicability in biological samples. On this basis, an “AND” logic gate was successfully constructed. Superior sensitivity to ATP in the presence of exonuclease was reflected, which greatly enhanced the system’s fluorescence. Importantly, the fluorescence sensing application of this nanomaterial inspired other target detection and enriched the building blocks of fluorescence sensing platform.

Nucleoside and nucleotide analogues are essential antivirals in the treatment of infectious diseases such as human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV), herpes simplex virus (HSV), varicella-zoster virus (VZV), and human cytomegalovirus (HCMV). To celebrate the 80th birthday of Prof. Dr. Erik De Clercq on 28 March 2021, this review provides an overview of his contributions to eight approved nucleos(t)ide drugs: (i) three adenosine nucleotide analogues, namely tenofovir disoproxil fumarate (Viread®) and tenofovir alafenamide (Vemlidy®) against HIV and HBV infections and adefovir dipivoxil (Hepsera®) against HBV infections; (ii) two thymidine nucleoside analogues, namely brivudine (Zostex®) against HSV-1 and VZV infections and stavudine (Zerit®) against HIV infections; (iii) two guanosine analogues, namely valacyclovir (Valtrex®, Zelitrex®) against HSV and VZV and rabacfosadine (Tanovea®-CA1) for the treatment of lymphoma in dogs; and (iv) one cytidine nucleotide analogue, namely cidofovir (Vistide®) for the treatment of HCMV retinitis in AIDS patients. Although adefovir dipivoxil, stavudine, and cidofovir are virtually discontinued for clinical use, tenofovir disoproxil fumarate and tenofovir alafenamide remain the most important antivirals against HIV and HBV infections worldwide. Overall, the broad-spectrum antiviral potential of nucleos(t)ide analogues supports their development to treat or prevent current and emerging infectious diseases worldwide.

Under-five years of age is a critical period for children’s growth and development. Nutritional deficiency during this period is associated with wasting, underweight and stunting. We aimed to conduct an epidemiological study using data derived from the GBD2019 to found the global distribution and changing trends of nutritional deficiencies among children under 5 years old, as well as the correlation between social development status and nutritional deficiencies. Nutritional deficiencies in children under 5 years has been substantially improved in the past decade; however, the progress has been unevenly distributed globally. The incidence and DALY rate decreased with the increase of socio-demographic index. In 2019, the incidence (51,872.0 per 100,000) was highest in Central Sub-Saharan Africa and the DALY rate (5597.1 per 100,000) was the highest in Western Sub-Saharan Africa. Among five subcategories of nutritional deficiencies in children under 5 years, vitamin A deficiency accounted for the largest proportion of incident cases (100,511,850, 62.1% in 2019), while the proportion of DALYs caused by protein–energy malnutrition was the highest (9,925,276, 62.0%). Nutritional deficiency in some countries remains worrisome, for whom policies guarantees and sustained efforts to control nutritional deficiencies are urgently needed.

A sequential [3 + 2]/[2 + 1] annulation domino reaction of crotonate-derived sulfur ylides and Morita-Baylis-Hillman carbonates of isatins for the construction of oxospiro[bicyclo[3.1.0]hexane-6,3'-indolin] scaffolds in moderate to good yields with almost 1:1 diastereoselectivity has been developed. Mild reaction conditions and readily accessible starting materials as well as excellent functional group compatibility render this transformation a powerful tool for the synthesis of spirocyclopropyloxindoles. A gram-scale synthetic procedure was also successfully carried out and a plausible reaction mechanism could be proposed.

Heat stress affects the physiology and production performance of Chinese Holstein dairy cows. As such, the selection of heat tolerance in cows and elucidating its underlying mechanisms are vital to the dairy industry. This study aimed to investigate the heat tolerance associated genes and molecular mechanisms in Chinese Holstein dairy cows using a high-throughput sequencing approach and bioinformatics analysis. Heat-induced physiological indicators and milk yield changes were assessed to determine heat tolerance levels in Chinese Holstein dairy cows by Principal Component Analysis method following Membership Function Value Analysis. Results indicated that rectal temperature (RT), respiratory rate (RR), and decline in milk production were significantly lower (p &lt, 0.05) in heat tolerant (HT) cows while plasma levels of heat shock protein (HSP: HSP70, HSP90), and cortisol were significantly higher (p &lt, 0.05) when compared to non-heat tolerant (NHT) Chinese Holstein dairy cows. By applying RNA-Seq analysis, we identified 200 (81 down-regulated and 119 up-regulated) significantly (|log2fold change| &ge, 1.4 and p &le, 0.05) differentially expressed genes (DEGs) in HT versus NHT Chinese Holstein dairy cows. In addition, 14 of which were involved in protein&ndash, protein interaction (PPI) network. Importantly, several hub genes (OAS2, MX2, IFIT5 and TGFB2) were significantly enriched in immune effector process. These findings might be helpful to expedite the understanding for the mechanism of heat tolerance in Chinese Holstein dairy cows.

Cholesterol concentration is one of the most significant indicators to assess health status. Lanthanide metal-organic framework (Ln-MOF) has a unique luminescence mechanism which makes it possess great potential in monitoring the variation of cholesterol. Thus, on the basic of intramolecular charge transfer theory, quenching theory, we aim to create a metal-organic framework (MOF) doped by europium to respond ratiometrically to the cholesterol in serum. The successful synthesis and the sensing mechanism of fluorescent probe were explained in detail by cross-corroborating by experiment and calculation and the high sensitivity of this fluorescent sensor is explained at the molecular level via DFT calculations. Under the excitation wavelength of 295 nm, the emission peaks at 614.5 nm and 532 nm, corresponding to Eu3+ and MIL-53(Fe), appeared quenching and enhancement respectively, and the ratiometric probes are constructed with favorable linearity. The probe has an ultrasensitive (limit of detection 0.33 mmol/L), significant stability and selectivity in response to the alteration of cholesterol. To investigate the emission law, by combining theoretical calculations and experimental results, a possible mechanism is proposed, in particular, the density of states and partial density of states were used to explain the internal electron migration of the probe when cholesterol interacts with it and the possible mechanism leading to quenching. The results showed the electron transfer efficiency of MIL-53(Fe) was enhanced after Eu3+ doping, which enhanced response sensitivity to the alternation of cholesterol. Meanwhile, based on the rules of electron arrangement outside the nucleus, it is proposed that the preferential filling arrangement of electrons in the transition from excited to ground state will lead to fluorescence quenching at Eu center. We anticipate our research to be an emerging method in cholesterol clinical tests and have further development in all walks of life.

A new carrier containing three hydrophobic octadecyls was employed to improve the yield and purity of the synthesized oligosaccharides, and the applicability of hydrophobic carrier for HASP (hydrophobically assisted switching phase) was also explored. High performance liquid chromatography (HPLC) was used to analyze the purity of oligosaccharides and improve the elution efficiency. Elution system was optimized and replaced by isopropyl alcohol and water, in which the excess donor and other impurities had a better solubility than that in methanol.

Gold and uranium mining has resulted in elevated concentrations of As and U contaminants in groundwater and soil, which has become a paramount concern for the environment and public health. Nanocleaning technology is one of the few permanent treatment alternatives to remove metal contaminants from soils. In recent years, nano-sized FeS, a washing solution that uses nanocleaning technology, has elicited extensive interest in environmental remediation because it does not produce secondary pollution. In this study, As and U were removed from contaminated soil by employing nano-sized FeS washing solution biologically synthesized by the iron-reducing bacteria Acidiphilium cryputum JF-5 and sulphate-reducing bacteria Desulfovibrio vulgaris miyazaki SRB. After cleaning with a nano-sized FeS suspension solution, culture-medium and pure-water for three cycles, the concentration of As in the nano-sized FeS experiment was higher than that of U, suggesting that the cleaning effect for As was better than that for U. The As and U removal rates of the nano-sized FeS cleaning experiment were 12% and 1.59%, respectively, over the three cycles. By using the sequential extraction method, the nano-sized FeS cleaning experiments mainly removed the As fraction bound to Fe/Al oxides. An ATR-FTIR spectral analysis indicated that the band observed at 461 cm−1 was attributed to the ν 4 symmetric stretching mode of AsO 4 3 − . The obvious vibrations observed at 798 cm−1 and 778 cm−1, were assigned to the ν 1 vibration of AsO 3 3 − . The band at 834 cm−1 was assigned to the weak vibration of UO2 while the band at 949 cm−1 was attributed to the Fe–O–D bending vibration. The results of this study provide helpful information and references for designing long-term and efficient bioremediation strategies for As- and U-containing soils.

Previously we reported that gene expression of astrocytic 5-HT

A modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) method using multi-walled carbon nanotubes (MWCNTs) as sorbent solvent combined with HPLC was developed for the simultaneous determination of olaquindox, carbadox, mequindox, quinocetone, and cyadox in animal feeds. Sample preparation was performed by ultrasound-assisted extraction (UAE) with methanol-acetonitrile-H2O (50:25:25, v/v/v) as extraction solvent and a final clean-up using QuEChERS with MWCNTs (30–50 nm) as the sorbent solvent. To achieve the optimum conditions for extraction the target analytes and modified QuEChERS toward target analytes, several parameters, including extraction solvent and time as well as the type and amount of the adsorbents were investigated. The detection and quantification of five quinoxalines were performed by HPLC-UV. The chromatographic separation was performed on a C18 column using methanol-water as the mobile phase. The limits of detection and quantification of the analytes ranged from 0.2 to 0.4 and 1.0 to 1.5 mg/kg, respectively. The recoveries from spiked sample ranged from 66.0 to 95.6% with the relative standard deviations of less than 14.1%. The developed method was successfully utilized to monitor real samples, demonstrating that it is simple, fast, and robust, and may be used to monitor quinoxalines residues in animal feeds.

Natural gas hydrate, which forms and agglomerates under the condition of low temperature and high pressure, can easily affect production or even lead to serious accidents. In order to prevent hydrate agglomeration in a pipeline, the industries generally add seriously excess inhibitor with experience, which causes serious environmental pollution and huge costs. However, due to the uncontrollable production and environmental conditions, hydrate plugging in a pipeline still occurs. According to the issues above, the method, for hydrate agglomeration online monitoring in a natural gas pipeline based on acoustic active excitation, has been presented in this paper. The method is to monitor and locate the hydrate agglomeration in a pipeline in order to take measures in time. Both the modeling and the experimental results show that the method can monitor and locate the hydrate agglomeration online at different locations with good accuracy.

Curcumin, a phytochemical agent in the spice turmeric, has received increasing attention for its anticancer, anti-inflammatory and antioxidant properties. However, application of curcumin has been limited due to its insolubility in water and poor bioavailability both clinically and experimentally. In addition, the protective effects and mechanisms of curcumin in eye diseases have been poorly studied. In the present study, we synthesized a curcumin analog, 1, 5-bis (2-trifluoromethylphenyl)-1, 4-pentadien-3-one (C3), which displayed improved protective effect against acrolein-induced toxicity in a human retinal pigment epithelial cell line (ARPE-19). At 5 μM, curcumin completely protected against acrolein-induced cell oxidative damage and preserved GSH levels and mitochondrial function. Surprisingly, C3 displayed a complete protective effect at 0.5 μM, which was much more efficient than curcumin. Both 0.5 μM C3 and 5 μM curcumin induced Nrf2 nuclear translocation and Nrf2 target genes transcription similarly. Experiments using Nrf2 siRNA showed that the protective effects of curcumin and C3 were eliminated by Nrf2 knockdown. Additionally, both curcumin and C3 activated the PI3/Akt pathway, however, Nrf2 activation was independent of this pathway, and therefore, we hypothesized that both curcumin and C3 activated phase II enzymes via directly disrupting the Nrf2/Keap1 complex and promoting Nrf2's nuclear translocation. Since acrolein challenge of ARPE-19 cells has been used as a model of smoking and age-related macular degeneration (AMD), we concluded that the curcumin analog, C3, may be a more promising drug candidate for its potential application for the prevention and treatment of eye diseases, such as AMD.

The current best serum marker for pancreatic cancer, CA 19-9, detects a carbohydrate antigen on multiple protein carriers. Better knowledge of the protein carriers of the CA 19-9 antigen in various disease states may lead to improved diagnostic tests. To identify proteins that carry the CA 19-9 antigen, we immunoprecipitated the CA 19-9 antigen from pooled sera and identified the associated proteins using mass spectrometry. Among the high-confidence identifications, we confirmed the presence of the CA 19-9 antigen on Apolipoprotein B-100 by antibody arrays and Western blot and on kininogen, ARVCF, and Apolipoprotein E by antibody arrays. We characterized the frequency and levels of the CA 19-9 antigen on the four proteins across various patient groups (pancreatic cancer, pancreatitis, and healthy controls) using antibody arrays. 10–25% of the subjects showed elevations of the antigen on each protein, but the elevations were not associated with disease state or total CA 19-9 levels. These results contribute to our knowledge of the carrier proteins of an important functional glycan and the rate at which the glycan is displayed. This work also demonstrates a strategy for using the complementary methods of mass spectrometry and antibody microarrays to identify protein carriers of glycans and assess the diagnostic value of measuring glycans on individual proteins.

Cystic lesions of the pancreas are increasingly being recognized due to the widespread use of high resolution abdominal imaging. Since certain cyst types are precursors to invasive cancer, this situation presents an opportunity to intervene prior to malignant progression. Effective implementation of that strategy has been hampered by difficulties in clearly distinguishing cystic lesions with no malignant potential from those with malignant potential. Here we explored whether glycosylation variants on specific proteins in cyst fluid samples could serve as biomarkers to aid in this diagnosis.We used a novel antibody-lectin sandwich microarray method to measure the protein expression and glycosylation of mucin (MUC)1, MUC5AC, MUC16, carcinoembryonic antigen, and other proteins implicated in pancreatic neoplasia in cyst fluid samples. Fifty-three cyst fluid samples were obtained from patients with mucinous cystic neoplasms (n=17), intraductal papillary mucinous neoplasms (n=15), serous cystadenomas (n=12), or pseudocysts (n=9), with confirmation of histologic diagnosis at surgical resection.The detection of a glycan variant on MUC5AC using the lectin wheat-germ agglutinin discriminated mucin-producing cystic tumors (mucinous cystic neoplasms+intraductal papillary mucinous neoplasms) from benign cystic lesions (serous cystadenomas+pseudocysts) with a 78% sensitivity at 80% specificity, and when used in combination with cyst fluid CA 19-9 gave a sensitivity of 87% at 86% specificity. These biomarkers performed better than cyst fluid carcinoembryonic antigen (37%/80% sensitivity/specificity).These results demonstrate the value of glycan variants for biomarker discovery and suggest that these biomarkers could greatly enhance the accuracy of differentiating pancreatic cystic tumors. Validation studies will be required to determine the clinical value of these markers.

Glycan arrays have enabled detailed studies of the specificities of glycan-binding proteins. A challenge in the interpretation of glycan array data is to determine the specific features of glycan structures that are critical for binding. To address this challenge, we have developed a systematic method to interpret glycan array data using a motif-based analysis. Each glycan on a glycan array is classified according to its component sub-structures, or motifs. We analyze the binding of a given lectin to each glycan in terms of the motifs in order to identify the motifs that are selectively present in the glycans that are bound by the lectin. We compared two different methods to calculate the identification, termed intensity segregation and motif segregation, for the analysis of three well-characterized lectins with highly divergent behaviors. Both methods accurately identified the primary specificities as well as the weaker, secondary specificities of all three lectins. The complex binding behavior of wheat germ agglutinin was reduced to its simplified, independent specificities. We compiled the motif specificities of a wide variety of plant lectins, human lectins, and glycan-binding antibodies to uncover the relationships among the glycan-binding proteins and to provide a means to search for lectins with particular binding specificities. This approach should be valuable for rapidly analyzing and using glycan array data, for better describing and understanding glycan-binding specificities, and as a means to systematize and compare data from glycan arrays.

Microarrays have been extremely useful for investigating binding interactions among diverse types of molecular species, with the main advantage being the ability to examine many interactions using small amount of samples and reagents. Microarrays are increasingly being used to advance research in the field of glycobiology, which is the study of the nature and function and carbohydrates in health and disease. Several types of microarrays are being used in the study of glycans and proteins in glycobiology, including glycan arrays to study the recognition of carbohydrates, lectin arrays to determine carbohydrate expression on purified proteins or on cells, and antibody arrays to examine the variation in particular glycan structures on specific proteins. This review will cover the technology and applications of these types of microarrays, as well as their use for obtaining complementary information on various aspects of glycobiology.

The natural gas vehicle market is rapidly developing throughout the world, and the majority of such vehicles operate on compressed natural gas (CNG). However, most studies on the emission characteristics of CNG vehicles rely on laboratory chassis dynamometer measurements, which do not accurately represent actual road driving conditions. To further investigate the emission characteristics of CNG vehicles, two CNG city buses and two CNG coaches were tested on public urban roads and highway sections. Our results show that when speeds of 0–10 km/hr were increased to 10–20 km/hr, the CO 2 , CO, nitrogen oxide (NO x ), and total hydrocarbon (THC) emission factors decreased by (71.6 ± 4.3)%, (65.6 ± 9.5)%, (64.9 ± 9.2)% and (67.8 ± 0.3)%, respectively. In this study, The Beijing city buses with stricter emission standards (Euro IV) did not have lower emission factors than the Chongqing coaches with Euro II emission standards. Both the higher emission factors at 0–10 km/hr speeds and the higher percentage of driving in the low-speed regime during the entire road cycle may have contributed to the higher CO 2 and CO emission factors of these city buses. Additionally, compared with the emission factors produced in the urban road tests, the CO emission factors of the CNG buses in highway tests decreased the most (by 83.2%), followed by the THC emission factors, which decreased by 67.1%.

Different strategies have been proposed for the development of protein subunit vaccine candidates for tuberculosis (TB), which shows better safety than other types of candidates and the currently used Bacillus Calmette-Guerin (BCG) vaccine. In order to develop more effective protein subunits depending on the mechanism of cell-mediated immunity against TB, a polyprotein CTT3H, based on 5 immunodominant antigens (CFP10, TB10.4, TB8.4, Rv3615c, and HBHA) with CD8(+) epitopes of Mycobacterium tuberculosis, was constructed in this study. We vaccinated C57BL/6 mice with a TB subunit CTT3H protein in an adjuvant of dimethyldioctadecylammonium/monophosphoryl lipid A/trehalose 6,6'-dibehenate (DDA/MPL/TDB, DMT) liposome to investigate the immunogenicity and protective efficacy of this novel vaccine. Our results demonstrated that DMT liposome-adjuvanted CTT3H vaccine not only induced an antigen-specific CD4(+) Th1 response, but also raised the number of PPD- and CTT3H-specific IFN-γ(+) CD8(+) T cells and elicited strong CTL responses against TB10.4, which provided more effective protection against a 60 CFU M. tuberculosis aerosol challenge than PBS control and DMT adjuvant alone. Our findings indicate that DMT-liposome is an effective adjuvant to stimulate CD8(+) T cell responses and the DMT-adjuvanted subunit CTT3H vaccine is a promising candidate for the next generation of TB vaccine.

The endocytosis of transferrin receptor (TfR) has served as a model to study the receptor-targeted cargo delivery system for cancer therapy for many years. To accurately evaluate and optically measure this TfR targeting delivery in vitro, a CHO cell line with enhanced green fluorescent protein (EGFP)-tagged human TfR was established. A chimera of the hTfR and EGFP was engineered by fusing EGFP to the amino terminus of hTfR. Data were provided to demonstrate that hTfR-EGFP chimera was predominantly localized on the plasma membrane with some intracellular fluorescent structures on CHO cells and the EGFP moiety did not affect the endocytosis property of hTfR. Receptor internalization occurred similarly to that of HepG2 cells expressing wild-type hTfR. The internalization percentage of this chimeric receptor was about 81 ± 3% of wild type. Time-dependent co-localization of hTfR-EGFP and PE-conjugated anti-hTfR mAb in living cells demonstrated the trafficking of mAb-receptor complexes through the endosomes followed by segregation of part of the mAb and receptor at the late stages of endocytosis. The CHO-hTfR cells preferentially took up anti-hTfR mAb conjugated nanoparticles. This CHO-hTfR cell line makes it feasible for accurate evaluation and visualization of intracellular trafficking of therapeutic agents conjugated with transferrin or Abs targeting the hTfRs.

Although Bacillus Calmette-Guerin (BCG) vaccine confers protection from Mycobacterium tuberculosis infection in children, its immune protection gradually wanes over time, and consequently leads to an inability to prevent the reactivation of latent infection of M. tuberculosis. Therefore, improving BCG for better control of tuberculosis (TB) is urgently needed. We thus hypothesized that recombinant BCG overexpressing immunodominant antigens expressed at different growth stages of M. tuberculosis could provide a more comprehensive protection against primary and latent M. tuberculosis infection. Here, a novel cocktail of recombinant BCG (rBCG) strains, namely ABX, was produced by combining rBCG::85A, rBCG::85B, and rBCG::X, which overexpressed respective multistage antigens Ag85A, Ag85B, and HspX of M. tuberculosis. Our results showed that ABX was able to induce a stronger immune protection than individual rBCGs or BCG against primary TB infection in C57BL/6 mice. Mechanistically, the immune protection was attributed to stronger antigen-specific CD4(+) Th1 responses, higher numbers of IFN-γ(+) CD4(+) TEM and IL-2(+) CD8(+) TCM cells elicited by ABX. These findings thus provide a novel strategy for the improvement of BCG efficacy and potentially a promising prophylactic TB vaccine candidate, warranting further investigation.

The antibody-lectin sandwich arrays (ALSA) is a powerful new tool for glycoproteomics research. ALSA enables precise measurements of the glycosylation states of multiple proteins captured directly from biological samples. The platform can be used in a high-throughput mode with low sample consumption, making it well suited to biomarker research exploring glycan alterations on specific proteins. This article provides detailed descriptions of the use of ALSA, with a particular focus on biomarker research. The preparation and selection of antibodies and lectins, the preparation and use of the arrays and samples, and special considerations for using the platform for biomarker research are covered.

Changes to the glycan structures of proteins secreted by cancer cells are known to be functionally important and to have potential diagnostic value. However, an exploration of the population variation and prevalence of glycan alterations on specific proteins has been lacking because of limitations in conventional glycobiology methods. Here we report the use of a previously developed antibody-lectin sandwich array method to characterize both the protein and glycan levels of specific mucins and carcinoembryonic antigen-related proteins captured from the sera of pancreatic cancer patients (n = 23) and control subjects (n = 23). The MUC16 protein was frequently elevated in the cancer patients (65% of the patients) but showed no glycan alterations, whereas the MUC1 and MUC5AC proteins were less frequently elevated (30 and 35%, respectively) and showed highly prevalent (up to 65%) and distinct glycan alterations. The most frequent glycan elevations involved the Thomsen-Friedenreich antigen, fucose, and Lewis antigens. An unexpected increase in the exposure of alpha-linked mannose also was observed on MUC1 and MUC5ac, indicating possible N-glycan modifications. Because glycan alterations occurred independently from the protein levels, improved identification of the cancer samples was achieved using glycan measurements on specific proteins relative to using the core protein measurements. The most significant elevation was the cancer antigen 19-9 on MUC1, occurring in 19 of 23 (87%) of the cancer patients and one of 23 (4%) of the control subjects. This work gives insight into the prevalence and protein carriers of glycan alterations in pancreatic cancer and points to the potential of using glycan measurements on specific proteins for highly effective biomarkers.

The CA 19-9 assay detects a carbohydrate antigen on multiple protein carriers, some of which may be preferential carriers of the antigen in cancer. We tested the hypothesis that the measurement of the CA 19-9 antigen on individual proteins could improve performance over the standard CA 19-9 assay. We used antibody arrays to measure the levels of the CA 19-9 antigen on multiple proteins in serum or plasma samples from patients with pancreatic adenocarcinoma or pancreatitis. Sample sets from three different institutions were examined, comprising 531 individual samples. The measurement of the CA 19-9 antigen on any individual protein did not improve upon the performance of the standard CA 19-9 assay (82% sensitivity at 75% specificity for early-stage cancer), owing to diversity among patients in their CA 19-9 protein carriers. However, a subset of cancer patients with no elevation in the standard CA 19-9 assay showed elevations of the CA 19-9 antigen specifically on the proteins MUC5AC or MUC16 in all sample sets. By combining measurements of the standard CA 19-9 assay with detection of CA 19-9 on MUC5AC and MUC16, the sensitivity of cancer detection was improved relative to CA 19-9 alone in each sample set, achieving 67-80% sensitivity at 98% specificity. This finding demonstrates the value of measuring glycans on specific proteins for improving biomarker performance. Diagnostic tests with improved sensitivity for detecting pancreatic cancer could have important applications for improving the treatment and management of patients suffering from this disease.