A Bifunctional Luminescent Whitening and Sensing Material Based on Photoluminescence and Mechanoluminescence.

A bifunctional luminescent whitening and luminescent sensing composite material, BaMgAl12O17:Eu2+/polydimethylsiloxane (BAM/PDMS), that utilizes natural sunlight and mechanical energy is presented. By increasing the Eu2+ content, the photoluminescence (PL) excitation spectrum of the material shows a maximum redshift of 23 nm due to 5d level splitting of Eu2+, resulting in more spectral overlap with sunlight and an excellent PL whitening effect. Meanwhile, the self-recoverable mechanoluminescence (ML) of the material can be easily excited under mechanical stimuli due to contact electrification, exhibiting a unique stress sensing effect. Based on the unique features of PL whitening and ML sensing, the material is applied to model cars through a spray process, and the results demonstrate that the bifunctional BAM/PDMS material shows promising applications in automobile decoration.

Deciphering Multidrug-Resistant Plasmids in Disinfection Residual Bacteria from a Wastewater Treatment Plant.

Current disinfection processes pose an emerging environmental risk due to the ineffective removal of antibiotic-resistant bacteria, especially disinfection residual bacteria (DRB) carrying multidrug-resistant plasmids (MRPs). However, the characteristics of DRB-carried MRPs are poorly understood. In this study, qPCR analysis reveals that the total absolute abundance of four plasmids in postdisinfection effluent decreases by 1.15 log units, while their relative abundance increases by 0.11 copies/cell compared to investigated wastewater treatment plant (WWTP) influent. We obtain three distinctive DRB-carried MRPs (pWWTP-01-03) from postdisinfection effluent, each carrying 9-11 antibiotic-resistant genes (ARGs). pWWTP-01 contains all 11 ARGs within an ∼25 Kbp chimeric genomic island showing strong patterns of recombination with MRPs from foodborne outbreaks and hospitals. Antibiotic-, disinfectant-, and heavy-metal-resistant genes on the same plasmid underscore the potential roles of disinfectants and heavy metals in the coselection of ARGs. Additionally, pWWTP-02 harbors an adhesin-type virulence operon, implying risks of both antibiotic resistance and pathogenicity upon entering environments. Furthermore, some MRPs from DRB are capable of transferring and could confer selective advantages to recipients under environmentally relevant antibiotic pressure. Overall, this study advances our understanding of DRB-carried MRPs and highlights the imminent need to monitor and control wastewater MRPs for environmental security.

[Association of serum irisin levels with glucose and lipid metabolism among adolescents in Yinchuan City from 2017 to 2020].

OBJECTIVE: To explore the relationship between serum irisin levels and glucose and lipid metabolism among adolescents in Yinchuan City. METHODS: From 2017 to 2020, a conbination of convenient sampling and stratified cluster random sampling method were used to select 1219 adolescents aged 12 to 18 years old in Yinchuan City as research subjects. The height and weight were measured using the height and sitting height meter and the bioelectrical impedance analyzer. Blood indicators such as fasting plasma glucose(FPG), totalcholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C) and high-density lipoprotein cholesterol(HDL-C) were measured using fully automatic biochemical analyzer. Serum irisin levels were measured by enzyme-linked immunosorbent assay(ELISA). Binary logistic regression was used to analyze the correlation between irisin and abnormal glucose and lipid metabolism. RESULTS: The FPG, TC, HDL-C and LDL-C levels of subjects in the highest tertile of irisin levels were significantly lower than those of subjects in the lowest tertile of irisin levels(F values were 5.13, 3.15, 3.07 and 5.01, P<0.05), and the differences were statistically significant(all P<0.05). The serum irisin levels in the hyperglycemia group(t=2.87, P<0.01), hypercholesterolemia group(t=2.36, P=0.02) and hyperLDL-Cemia group(t=2.34, P=0.02) were significantly lower than those in the normoglycemia group, normal TC group and normal LDL-C group. Meanwhile, the irisin level in the low HDL-Cemia group(t=-2.57, P=0.01) was significantly higher than that in the normal HDL-C group, and the differences were statistically significant(P<0.05). Participants in the highest tertile of irisin had 0.51, 0.49 and 0.50 times the risk of hyperglycemia(OR=0.51, 95%CI 0.29-0.87), hypercholesterolemia(OR=0.49, 95%CI 0.27-0.89) and hyperLDL-cemia(OR=0.50, 95%CI 0.25-0.99) compared with those in the lowest tertile. CONCLUSION: Low levels of irisin are associated with the occurrence of hyperglycemia, hypercholesterolemia, and hyperLDL-Cemia in adolescents.

Intra-Individual Variation in Disease-Specific IgG Fc Glycoform Ratios to Monitor the Disease Progression of Lung Cancer.

Aberrant protein glycosylation is an active pathological alteration related to the progression of cancers. The speed of progression varies among individuals, increasing the difficulties of prognosis assessment. Hence, evaluating variation in glycosylation using patients themselves as their own controls is a potential way to reduce the impact of individual differences on progression monitoring. Here, following a longitudinal follow-up study involving 125 lung cancer (LC) patients with progressive disease, we isolated disease-specific IgG from serum using polyacrylamide gel electrophoresis, obtained IgG glycoform ratios using mass spectrometry, and then set a fold-change cutoff of 1.5 to utilize the intra-individual variation in IgG glycosylation to monitor PD. We found that the serial monitoring of 15 types of glycoform ratios provided an effective way for monitoring LC progression. Over 1.5-fold changes in glycoform ratios relative to the first observed value were detected in 117 of 125 LC patients (93.6%). Our established method predicted LC progression 55.8 (IQR 31.1-90.1) weeks earlier than imaging examination did. In summary, intra-individual variation in IgG glycoform ratios is useful to monitor LC progression, expanding our knowledge about the relationship between IgG glycosylation and cancer prognosis. The raw data files are available via the ProteomeXchange Consortium with the identifier PXD037541.

Clinical characteristics and risk factors of cerebral cavernous malformation-related epilepsy.

PURPOSE: This study aimed to summarize the clinical characteristics and explore the risk factors for cerebral cavernous malformation (CCM)-related epilepsy (CRE). METHODS: We retrospectively analyzed the clinical data of patients with CCM in our cerebral vascular malformations database. Descriptive statistics were used to present the clinical characteristics of CRE patients. Patients were divided into a CRE and a non-CRE group according to clinical presentation. Binary logistic regression analysis was used to analyze the risk factors of CRE. RESULTS: A total of 199 patients with CCM confirmed by postoperative pathological examination were enrolled, 93 of whom were diagnosed with CRE, and 34 patients had drug-resistant epilepsy. The most common seizure type of CRE patients was focal to bilateral tonic-clonic seizure (FBTCS), followed by focal impaired awareness motor seizure. All CCM lesions were supratentorial, 97.8% of which involved the cerebral cortex, 86.0% of lesions had hemosiderin rim, and 50.5% of lesions were located in the temporal lobe. Binary logistic regression analysis indicated that CCM diagnosis age ≤ 44 years (odds ratio [OR] 2.79, p = 0.010), temporal lobe lesion location (OR = 9.07, p = 0.042), medial temporal lobe lesion (OR = 14.09, p = 0.002), cortical involvement of the lesion (OR = 32.77, p = 0.010), and hemosiderin rim around the lesion (OR = 16.48, p = 0.001) significantly increased the risk of CRE. CONCLUSIONS: The most common seizure type of CRE was FBTCS. Those whose CCM diagnosis age was ≤ 44 years, having a temporal lobe lesion location, especially the medial temporal lobe lesion, cortical involvement, and hemosiderin rim around the lesion had a higher risk of developing CRE.

The relationship between different metabolic phenotypes and bone indices in Chinese children and adolescents aged 12-18 years old.

Data regarding different metabolic phenotypes and bone markers including bone mineral content (BMC) and osteocalcin (OCN) among children and adolescents are very limited. Hence, the purpose of this investigation was to explore the relationship between different metabolic phenotypes and BMC or OCN among Chinese children and adolescents. This cross-sectional study included 1,328 children and adolescents aged between 12 and 18 years who were selected from four schools in Yinchuan city from 2018 to 2020 by stratified cluster random sampling. Subjects were divided into four groups according to BMI and metabolic status, as follows: metabolically healthy obesity (MHO), metabolically unhealthy obesity (MUO), metabolically unhealthy normal weight (MUNW), and metabolically healthy normal weight (MHNW). The MHNW, MUNW, MHO, and MUO phenotypes in boys were 48.4%, 30.5%, 6.7%, and 14.4%, respectively, and were 47.8%, 33.6%, 6.6%, and 12.1% in girls, respectively. The MHO and MUO phenotypes had higher BMC than the MHNW or MUNW phenotype (all p < 0.05), and the MUO phenotype with BMC was significantly higher than MHO group in boys (p < 0.05). We discovered a significant positive correlation between BMC and the MHO (OR = 8.82, 95% CI = 2.04-38.16), MUO phenotypes (OR = 13.53, 95% CI = 4.10-44.70), while no association was found between OCN and metabolic phenotypes in neither boys nor girls. Overweight/obese children and adolescents had higher BMC, and there existed sex differences in the effect of metabolic status on BMC among them. OCN was not supposed to be an index of bone health in this study.

Proteomics and post-translational modifications analysis of umbilical mesenchymal stem cells aging.

Mesenchymal stem cells (MSCs) are considered to be critical for regenerative medicine. However, long-term culturing of MSCs will induce aging of MSCs, and thereafter impair cellular function. Changes in proteomics have been reported to be involved in cell aging, and therefore investigations on cell aging of MSCs at levels of proteomics and post-translational protein modifications (PTM) are ultimately important. In the present study, human umbilical cord mesenchymal stem cells (hUMSCs) were exposed to different culture conditions for either 7 or 30 days. Proteins changes during cell culture were investigated using tandem mass tag (TMT) labeling quantitative approach, and N-glycosylation patterns were analyzed using multistage mass spectrometry. We identified 66 proteins (fold change >1.50) that were differentially expressed in long-term culture. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that aging exerts a side effect on hUMSCs by affecting various molecular functions and biological processes, such as lysosome, autophagy and post-translational protein modification. Glycosylation analysis indicates that cell N-glycan patterns are associated with aging of MSCs. Our results presented here should contribute to future studies on cell aging and cellular quality controls related to MSCs as regenerative medicine.

Advanced Luminescence Anticounterfeiting Based on Dynamic Photoluminescence and Non-Pre-Irradiation Mechanoluminescence.

Luminescence anticounterfeiting is one of the most significant technologies to protect information security. However, the luminescence of the present anticounterfeiting logo is static, which is easily counterfeited by substitutes, and it always requires an ultraviolet lamp in use, which is inconvenient in application. In this work, according to the present deficiencies of luminescence anticounterfeiting, an interesting phosphor CaZnGe2O6/Mn2+ with unique features of dynamic photoluminescence and non-pre-irradiation mechanoluminescence is developed for the first time. The photoluminescence color of the phosphor can dynamically change from green to red during irradiation, and the non-pre-irradiation mechanoluminescence of the phosphor-based elastomer can be easily stimulated by mechanics such as stretching, bending, or scratching with a finger. By combining the two features of the CaZnGe2O6/Mn2+ phosphor, an advanced dual-mode luminescence anticounterfeiting is designed, and a luminescence logo is fabricated for the anticounterfeiting test. The result demonstrates that this advanced luminescence anticounterfeiting based on the phosphor is not only safer but also more convenient in application.

Silencing of circCDC14A prevents cerebral ischemia-reperfusion injury via miR-23a-3p/CXCL12 axis.

Ischemic stroke is one of the main causes of death and disability. Circular RNAs (circRNAs) have received extensive attention in the pathogenesis of ischemic stroke. Here, we evaluated the role of circCDC14A in cerebral ischemia-reperfusion (CI/R) injury in vivo and in vitro. The expression of circCDC14A was significantly upregulated in the middle cerebral artery occlusion (MCAO) model and oxygen and glucose deprivation/reoxygenation (OGD/R)-treated HT22 cells. Knockdown of circCDC14A suppressed the cell viability reduction caused by OGD/R, as well as cell damage and apoptosis. Mechanistically, circCDC14A acted as a sponge for miR-23a-3p and promoted the expression of chemokine stromal-derived factor-1 (CXCL12) by negatively regulating miR-23a-3p. Rescue experiments further confirmed that miR-23a-3p inhibitor or circCDC14A-overexpression vectors blocked the beneficial effects of circCDC14A knockdown in OGD/R-induced HT22 cells. Moreover, knockdown of circCDC14A suppressed MCAO-induced cerebral infarction and neurological damage, as well as the brain tissue damage and neuronal apoptosis in vivo. Consistently, miR-23a-3p antagomir treatment abolished the cerebral protective effects of circCDC14A knockdown on MCAO mice. In conclusion, circCDC14A promoted CI/R injury by regulating the miR-23a-3p/CXCL12 axis, which suggested that circCDC14A may become a potential therapeutic target for CI/R injury.

Association between body composition and blood pressure in normal-weight Chinese children and adolescents.

BACKGROUND: The aim of this study was to assess the associations of body fat distribution and lean body mass (LBM) with blood pressure (BP) in normal-weight Chinese children and adolescents. METHODS: A total of 898 normal-weight Chinese children and adolescents, aged 10-18 years, were included this cross-sectional study via a cluster sampling method. The bioelectrical impedance analysis (BIA) was used to measure body composition. The participants were measured for blood pressure (BP) using a calibrated electronic sphygmomanometer according to the standard method by the "American Hypertension Education Project Working Group". RESULTS: Body composition was related to abnormal BP in normal-weight children and adolescents. After the model adjusted for age, smoking, and drinking, regression analysis showed that fat mass percentage (FMP) was negatively associated with abnormal BP, while LBM was positively associated with abnormal BP in boys(P < 0.05). Whereas FMP and visceral fat level (VFL) were positively associated with abnormal BP in girls (P < 0.05). CONCLUSIONS: There are sex differences in the relationships between total body fat, visceral fat and lean body mass with abnormal BP in normal-weight youths. Therefore, it is of great significance to pay attention to the relative influence of the body composition of the boys and girls in the prevention and treatment of hypertension in youths.

Gain-Phase Error-Calibrated Piezoelectric Sensor Array-Based Impact Localization on Stiffened Curved Composite Structures.

Stiffened structure-induced gain-phase errors degrade the performance of the high-resolution two-dimensional multiple signal classification (2D-MUSIC) algorithm, which makes it impossible to ensure the high accuracy of impact localization results. To eliminate the localization bias caused by these errors, a calibrated 2D-MUSIC-based impact localization method is first introduced. Firstly, time-frequency characteristics of the non-stationary impact signals are evaluated by experiment to obtain a clear first wave packet or a wave packet that purely corresponds to a single mode through continuous wavelet transform (CWT). Then, the uniform linear array covariance matrix with gain-phase errors is calibrated to be constructed as a Toeplitz structural matrix. By reconstructing covariance matrix R, 2D-MUSIC-based impact localization is calibrated for stiffened curved composite structures. Experimental research on the stiffened curved composite panel is carried out, and these impact localization results demonstrate the validity and effectiveness of the calibrated 2D-MUSIC-based method.

[Association of serum uric acid with cardiovascular risk factors and their clustering among children and adolescents in Yinchuan City].

OBJECTIVE: To investigate the relationship between serum uric acid and cardiovascular risk factors and cardiovascular risk factor clustering(CVRFC) in children and adolescents in Yinchuan, Ningxia. METHODS: A present study design was adopted, and 1486 children and adolescents aged 10-18 years in urban areas of Yinchuan City were selected as study subjects with a mean age of(14.3±1.4) years in 2015, 2017 to 2018 by stratified clustering sampling, including 728(49.0%) boys, 1157(77.9%) Han, 170(11.4%) Hui and 159(10.7%) from other ethnic groups. All study subjects completed questionnaires, physical measurements and biochemical tests. RESULTS: Hyperuricemia(HUA) was significantly positive associated with abdominal obesity(OR=3.23, 95%CI 2.37-4.40), hypertension(OR=1.64, 95%CI 1.21-2.23), dyslipidemia(OR=1.51, 95%CI 1.17-1.96), CVRFC≥2(OR=3.71, 95%CI 2.80-4.93) and CVRFC≥3(OR=6.92, 95% CI 4.18-11.64)(P<0.05). There was an additive interaction between HUA and gender on cardiovascular risk factors and their aggregation. Compared with non-HUA girls, HUA girls have 3.57 times(95%CI 2.26-5.64) the risk of abdominal obesity, dyslipidemia, CVRFC≥2 and CVRFC≥3, respectively, 1.65 times(95%CI 1.10-2.47), 4.10 times(95%CI 1.10-2.47) and 7.63 times(95%CI 3.67-15.89). HUA boys have 1.75 times(95%CI 1.16-2.65) the risk of abdominal obesity, dyslipidemia, CVRFC≥2 and CVRFC≥3, respectively, 2.14(95%CI 1.51-3.03) times, 4.27 times(95%CI 2.98-6.13) and 7.97 times(95%CI 4.11-15.44), (P<0.05). CONCLUSION: Hyperuricemia was significantly positive associated with cardiovascular risk factors and their aggregation in children and adolescents in Yinchuan, and there was an additive interaction between hyperuricemia and gender.

Signaling Lymphocytic Activation Molecule Family-7 Alleviates Corneal Inflammation by Promoting M2 Polarization.

BACKGROUND: Signaling lymphocytic activation molecule family-7 (SLAMF7) functions as an immune checkpoint molecule on macrophages in antitumor immunity. However, its role in bacterial infection remains largely unknown. METHODS: Bone marrow-derived macrophages (BMDMs) isolated from wild-type (WT) or SLAMF7 knockout (KO) mice were infected with bacteria or treated with lipopolysaccharide/interferon-γ to investigate the expression and function of SLAMF7 in macrophage polarization. A Pseudomonas aeruginosa keratitis murine model was established to explore the effect of SLAMF7 on P. aeruginosa keratitis using WT vs SLAMF7 KO mice, or recombinant SLAMF7 vs phosphate-buffered saline-treated mice, respectively. RESULTS: SLAMF7 expression was enhanced on M1-polarized or bacterial-infected macrophages, and infiltrating macrophages in P. aeruginosa-infected mouse corneas. SLAMF7 promoted M2 polarization by inducing STAT6 activation. In vivo data showed that SLAMF7 KO aggravated, while treatment with recombinant SLAMF7 alleviated, corneal inflammation and disease severity. In addition, blockage of M2 polarization by Arg-1 inhibitor abrogated the effect of recombinant SLAMF7 in disease progression. CONCLUSIONS: SLAMF7 expression in macrophages was induced upon M1 polarization or bacterial infection and alleviated corneal inflammation and disease progression of P. aeruginosa keratitis by promoting M2 polarization. These findings may provide a potential strategy for the treatment of P. aeruginosa keratitis.

Genome-wide identification of ubiquitin proteasome subunits as superior reference genes for transcript normalization during receptacle development in strawberry cultivars.

BACKGROUND: Gene transcripts that show invariant abundance during development are ideal as reference genes (RGs) for accurate gene expression analyses, such as RNA blot analysis and reverse transcription-quantitative real time PCR (RT-qPCR) analyses. In a genome-wide analysis, we selected three "Commonly used" housekeeping genes (HKGs), fifteen "Traditional" HKGs, and nine novel genes as candidate RGs based on 80 publicly available transcriptome libraries that include data for receptacle development in eight strawberry cultivars. RESULTS: The results of the multifaceted assessment consistently revealed that expression of the novel RGs showed greater stability compared with that of the "Commonly used" and "Traditional" HKGs in transcriptome and RT-qPCR analyses. Notably, the majority of stably expressed genes were associated with the ubiquitin proteasome system. Among these, two 26 s proteasome subunits, RPT6A and RPN5A, showed superior expression stability and abundance, and are recommended as the optimal RGs combination for normalization of gene expression during strawberry receptacle development. CONCLUSION: These findings provide additional useful and reliable RGs as resources for the accurate study of gene expression during receptacle development in strawberry cultivars.

Causal link between immunoglobulin G glycosylation and cancer: A potential glycobiomarker for early tumor detection.

Changes in immunoglobulin G (IgG) glycan structures are currently believed to closely related to the emergence of cancer. In this review, we summarize the current body of evidence suggesting that differences in serum IgG glycosylation patterns correspond to changes in multiple types of cancer. Modifications include IgG terminal N-link galactosylation, IgG core fucosylation, IgG terminal sialylation, and IgG terminal bisecting N-acetylglucosamine. IgG N-glycomic alterations represent promising novel biomarkers for non-invasive-cancer diagnosis, prognosis, and progression monitoring; they are characterized by high sensitivity and specificity, compensating for previously identified glycobiomarkers.

GPER1 Modulates Synaptic Plasticity During the Development of Temporal Lobe Epilepsy in Rats.

G-protein coupled estrogen receptor 1 (GPER1) is a novel type of estrogen receptor. Several studies have shown that it has an anti-inflammatory action,which plays an important role in remyelination and cognitive ability adjustment. However, whether it is involved in the development of temporal lobe epilepsy (TLE) is still unknown. The present study established a TLE model by intraperitoneal injection of lithium chloride (3 mmol/kg) and pilocarpine (50 mg/kg) in rats to study the effect of GPER1 in the synaptic plasticity during the development of temporal lobe epilepsy. A microinjection cannula was implanted into the lateral ventricle region of rats via a stereotaxic instrument. G-1 is the specific GPER1 agonist and G15 is the specific GPER1 antagonist. The G1 or G15 and Dimethyl sulfoxide were injected into the rat brains in the intervention groups and control group, respectively. After G1 intervention, the learning and memory abilities and hippocampal neuron damage in epileptic rats were significantly improved, while G15 weakened the neuroprotective effect of GPER1. Meanwhile, G1 controlled the abnormal formation of hippocampal mossy fiber sprouting caused by seizures, and participated in the regulation of synaptic plasticity by reducing the expression of Synapsin I and increasing the expression of gephyrin. Inhibitory synapse gephyrin may play a significant role in synaptic plasticity.

Fe3O4@PANI: a magnetic polyaniline nanomaterial for highly efficient and handy enrichment of intact N-glycopeptides.

Glycosylation of proteins plays important roles in the occurrence and development of chronic diseases. In this study, we report an enrichment method of intact N-glycopeptides using a magnetic polyaniline nanomaterial (Fe3O4@PANI). Under the synergistic effect of hydrogen bonding and electrostatic adsorption, Fe3O4@PANI can rapidly and easily enrich N-glycopeptides derived from standard protein (bovine fetuin and transferrin) tryptic digests and serum haptoglobin tryptic digests. Finally we have detected 63 glycopeptides in the glycosylation sites of both N204 and N211 from the serum haptoglobin beta chain using MALDI FTICR MS. Compared with non-magnetic materials, Fe3O4@PANI can achieve complete separation from complex biological samples, meeting the requirement of the high purity of samples for mass spectrometric detection. Overall, Fe3O4@PANI exhibits great application potential in the highly efficient enrichment of intact N-glycopeptides due to its stability and convenient preparation.

Best-first search guided multistage mass spectrometry-based glycan identification.

MOTIVATION: Glycan identification has long been hampered by complicated branching patterns and various isomeric structures of glycans. Multistage mass spectrometry (MSn) is a promising glycan identification technique as it generates multiple-level fragments of a glycan, which can be explored to deduce branching pattern of the glycan and further distinguish it from other candidates with identical mass. However, the automatic glycan identification still remains a challenge since it mainly relies on expertise to guide a MSn instrument to generate spectra. RESULTS: Here, we proposed a novel method, named bestFSA, based on a best-first search algorithm to guide the process of spectrum producing in glycan identification using MSn. BestFSA is able to select the most appropriate peaks for next round of experiments and complete the identification using as few experimental rounds. Our analysis of seven representative glycans shows that bestFSA correctly distinguishes actual glycans efficiently and suggested bestFSA could be used in practical glycan identification. The combination of the MSn technology coupled with bestFSA should greatly facilitate the automatic identification of glycan branching patterns, with significantly improved identification sensitivity, and reduce time and cost of MSn experiments. AVAILABILITY AND IMPLEMENTATION: http://glycan.ict.ac.cn. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Bovine serum albumin affects N-glycoforms of murine IgG monoclonal antibody purified from hybridoma supernatants.

Immunoglobulin G (IgG) is a class of monoclonal antibodies (mAbs) commonly produced in mammalian cell lines. These cell lines are grown in finely adjusted culture media, which contain components that may impact glycoforms. As variation of N-glycoforms can impact the biological properties of IgGs, medium composition should be controlled. Here, we studied the effects on IgG N-glycoforms of different components in hybridoma culture media, specifically compared bovine serum albumin (BSA) with other small molecules, using a matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight multistage mass spectrometry (MALDI-QIT-TOF MSn)-based approach. We show that small molecular additives caused little change in glycan species, though a number of these reagents, especially glutamine, affected levels of glycosylation. In comparison, the addition of macromolecular protein BSA significantly changed IgG N-glycan patterns, not only in species but also in glycosylation levels. Together, our finding suggests that BSA increases the complexity of IgG N-glycoforms, thus raising the difficulty in maintaining glycoforms consistency during antibody production. Therefore, the effect of BSA on IgG N-glycans should be considered when designing optimal medium formulations for IgG production. KEY POINTS: • Small molecular medium additives only affect glycosylation levels of IgG N-glycans. • BSA significantly changes IgG N-glycoforms as a medium additive. • BSA's skewing of IgG N-glycoforms should be considered in IgG production.

Prolactin regulates LAT1 expression via STAT5 (signal transducer and activator of transcription 5) signaling in mammary epithelial cells of dairy cows.

The l-type amino acid transporter 1 (LAT1; also known as SLC7A5) is a transporter that allows the uptake of large neutral amino acids into mammalian cells. In dairy cows, LAT1 is highly expressed in lactating mammary tissues and involved in milk protein synthesis. Prolactin (PRL) has a lactogenic role and is capable of inducing milk production in ruminants. However, the relationship between PRL stimulation and LAT1 expression in dairy cow mammary gland has not been well understood. In this study, we showed that PRL stimulation increased expression of LAT1 and ß-casein in mammary epithelial cells of dairy cows. The stimulatory effect of PRL on milk protein production was inhibited by LAT1-specific inhibitor or LAT1 knockdown, suggesting that PRL-induced milk protein production is involved in LAT1 expression. To determine whether the PRL signaling pathway participates in regulation of LAT1 expression, PRLR (PRL receptor) or STAT5 (signal transducer and activator of transcription 5) was knocked down by short interfering (si)RNA in mammary epithelial cells of dairy cows. Western blot results showed that knockdown of PRLR or STAT5 with siRNA markedly decreased PRL-stimulated LAT1 expression. In addition, we observed a marked increase in plasma membrane expression of LAT1 in PRL-stimulated cells compared with control cells. These observations indicated that PRL signaling can regulate LAT1 expression and activity in mammary epithelial cells of dairy cows, contributing to increased amino acid availability and milk protein synthesis in mammary gland of dairy cow.