Sustained Phosphorus Removal and Enrichment through Off-Flow Desorption in a Reservoir of Membrane Capacitive Deionization.

In this study, we used a membrane capacitive deionization device with a reservoir (R-MCDI) to enrich phosphorus (P) from synthetic wastewater. This R-MCDI had two small-volume electrode chambers, and most of the electrolyte was contained in the reservoir, which was circulated along the electrode chambers. Compared with conventional MCDI, R-MCDI exhibited a phosphate removal rate of 0.052 µmol/(cm2·min), approximately double that of MCDI. This was attributed to R-MCDI's utilization of OH- alternative adsorption to remove phosphate from the influent. Noticing that around 73.9% of the removed phosphate was stored in the electrolyte in R-MCDI, we proposed a novel off-flow desorption operation to enrich the removed phosphate in the reservoir. Exciting results from the multicycle experiment (∼8 h) of R-MCDI showed that the PO43--P concentration in the reservoir increased all the way from the initial 152 mg/L to the final 361 mg/L, with the increase in the P charge efficiency from 5.5 to 22.9% and the decrease in the energy consumption from 28.2 to 6.8 kW h/kg P. The P recovery performance of R-MCDI was evaluated by viewing other similar studies, which revealed that R-MCDI in this study achieved superior P enrichment with low energy consumption and that the off-flow desorption proposed here considerably simplified the operation and enabled continuous P enrichment.

Size distribution analysis of residual host cell DNA fragments in lentivirus by CGE-LIF.

During the production of cell and gene therapy products, residual host cell DNA (HCD) could cause safety risks of the biological products, and the longer the residual HCD fragment, the greater the risk to the human body. For this reason, it was necessary to develop an effective method for the size distribution analysis of residual HCD fragments with high accuracy and sensitivity. In this study, capillary gel electrophoresis with laser-induced fluorescence detector (CGE-LIF) was used to analyze the size distribution of residual HCD fragments in lentivirus products. The results confirmed that lentiviral RNA genome could interfere with the size distribution analysis of residual HCD fragments. By optimizing the amount of RNase I and digestion time in sample pretreatment process, the interfere of RNA genome could be avoided. The specificity, precision, accuracy, linear range, the detection of limit (LOD), and the quantification of limit (LOQ) of CGE-LIF method were also validated. The results showed that the CGE-LIF method had a good performance both in terms of specificity and reproducibility. The intra- and inter-day relative standard deviations of migration time and corrected peak area were all less than 1% and 2%, respectively. The 200 bp DNA marker had a good linearity between 50 and 1000 pg/ml. The LOD and LOQ of 200 bp DNA marker were 2.59 and 8.64 pg/ml, respectively. In addition, this method was successfully used to analyze the size distribution analysis of residual HCD fragments in lentivirus products with different production processes.

Intraocular Pressure and Myopia Progression, Axial Length Elongation in Rural Chinese Children.

SIGNIFICANCE: This study reported the relationship between intraocular pressure (IOP) and myopia progression, which helps to understand more comprehensively whether IOP can be an important reference factor to intervene in the progression of myopia. PURPOSE: This study aimed to investigate the association between IOP and myopia progression as well as axial length elongation in rural Chinese children. METHODS: A total of 598 (598 of 878 [68.1%]) children (6 to 17 years) from the baseline Handan Offspring Myopia Study who completed a 3.5-year follow-up vision examination were included. Ocular examinations at both visits included cycloplegic autorefraction, IOP, and axial length measurements. RESULTS: Children with myopia had the highest baseline IOP of the three refractive groups (14.13 ± 1.31, 13.78 ± 1.71, and 13.59 ± 1.64 mmHg in myopes, emmetropes, and hyperopes, respectively, P = .002). However, IOPs showed no significant difference between eyes with or without newly developed myopia (13.63 ± 1.68 vs. 13.89 ± 1.68, P = .16), with or without faster myopia progression (13.75 ± 1.61 vs. 13.86 ± 1.63, P = .46), or with axial length elongation (13.80 ± 1.61 vs. 13.76 ± 1.64, P = .80). The multivariate regression analysis demonstrated that neither baseline refractive error ( ß = -0.082, P = .13) nor baseline axial length ( ß = -0.156, P = .08) was associated with baseline IOP. CONCLUSIONS: Myopic eyes have slightly higher IOP compared with emmetropic and hyperopic eyes, although it was not clinically significant. However, IOP was not found to be associated with either myopia progression or axial length elongation in this cohort sample of rural Chinese children.

Effects of soybean hulls and corn stalk on the performance, colostrum composition and faecal microflora of pregnant sows.

This study was conducted to investigate the effects of different supplementation levels of soybean hulls and corn stalk in high-fibre gestation diet on the performance, colostrum composition and faecal microbiota of sows. Forty first-farrowing Danish Landrace sows were randomly assigned to five dietary treatment groups. The control (CON, 3.15% crude fibre) group was fed a normal diet, and the treatment groups were soybean hulls low-fibre (SHL, 6.00% crude fibre) group, soybean hulls high-fibre (SHH, 8.00% crude fibre) group, corn stalk low-fibre (CSL, 6.00% crude fibre) group and corn stalk high-fibre (CSH, 8.00% crude fibre) group. The weaning weight of the litter and the average daily feed intake of the lactating sows in the SHL, SHH and CSH groups were higher than those in the CON group (p < 0.05). The immunoglobulin A and G levels of the colostrum in the SHL, SHH, CSL and CSH groups were higher than those in the CON group (p < 0.05), and the immunoglobulin M levels in the SHL, SHH and CSH groups were higher than those in the CON group (p < 0.05). The abundance of Proteobacteria at the phylum level in the CON group was higher than that in the CSL, CSH and SHH groups (p < 0.05). The abundance of Lactobacillaceae at the family level in the SHH and CSL groups were higher than that in the CON group (p < 0.05). The abundance of Lactobacillus at the genus level in the SHH and CSL groups were higher than that in the CON group (p < 0.05). In conclusion, SHH group had the best effect, and the optimal crude fibre level in the gestation diet of sows is 8%.

A platform method for plasmid isoforms analysis by capillary gel electrophoresis.

In the production of novel biological products, plasmids are often engineered into delivery vectors for target genes, which can be used directly as vaccines or as intermediate products for gene/cell therapy. Plasmid DNA exists in several topological forms such as supercoiled, linear, and open circular. As supercoiled plasmid shows the highest efficiency in transfecting eukaryotic cells, the content of supercoiled plasmids becomes an important indicator of plasmid quality. CGE is an effective analysis method for separating different topological structures of plasmids. For the purpose of providing plasmid manufacturers and regulatory agencies with an efficient and readily used tool for monitoring the quality of plasmids, this article identifies the optimal separation and detection conditions of CGE, presents a platform-based plasmid analytical method, and uses plasmid of different sizes to verify the feasibility of this method. In terms of detector, the LIF detector has obvious advantages over the ultraviolet detector in sensitivity and resolution. Using the optimal CE condition (10× gel buffer), baseline separation of different topological forms and impurities can be achieved for different plasmid sizes (5.9, 7.8, 15.4 kb). In addition, 6.5 kb plasmid was used to compare the different separation technologies such as CGE-LIF, ion exchange chromatography and agarose gel electrophoresis. The result shows that CGE-LIF can provide better resolution and quantitation accuracy than ion exchange chromatography and agarose gel electrophoresis. CGE-LIF, as a quick and convenient method to separate and quantify plasmids, has the advantages of high sensitivity, high resolution, and high quantitative accuracy. Therefore, it is ideal for analysis of plasmids with different sizes, and it can also be used as a platform method for manufacturers and regulatory agencies to monitor the purity and stability of plasmids.

Cooling therapy upregulates protein C activation in heat stressed rats: An experimental study.

Protein C (PC) pathway homeostasis is implicated in heat stress (HS). This study determines whether cooling could improve the PC pathway in HS. Fifty-six anesthetized rats were warmed to achieve HS (rectal temperature [Tr] 42°C). These rats were divided into seven groups: (a) control group:sacrifice immediately 15 min after HS; (b) HS+I:sacrifice immediately after 15 min ice-water treatment or (c) 3 hr after HS; (d) HS+C:sacrifice immediately after 15-min cold-water treatment or (e) 3 hr after HS; (f) HS: sacrifice immediately 15 min after HS or (g) 3 hr after HS. Plasma PC, activated protein C (APC), and soluble thrombomodulin (sTM) levels were tested at both time points. After cooling, Tr in the HS+I and HS+C groups significantly decreased, when compared with the HS group, and Tr was significantly lower in the HS+I group than in the HS+C group ( p < 0.05). Furthermore, sTM levels were highest in the HS group among the groups at both time points. Plasma PC and APC levels increased after HS. In the HS+I and HS+C groups, plasma APC levels and the APC/PC ratio significantly increased at both time points. The proportions were significantly higher in the HS+I group than in the HS+C group, and there was no significant increase in APC/PC ratio in the HS group. Cooling exerts an anticoagulant effect following HS by increasing APC levels. Ice-water blanket therapy is more effective than cold-water blanket therapy in increasing APC levels.

A facile strategy to prepare (N, Ni, P) tri-doped echinus-like porous carbon spheres as advanced anode for lithium ion batteries.

Heteroatom doped carbon nanomaterials are often employed as advanced anodes for lithium ion batteries (LIBs) because of their stable structure, high capacity and low cost. In this work, we proposed a novel strategy to synthesize high density (N, Ni, P) tri-doped echinus-like porous carbon spheres (PCS) by carbonizing a metal-organophosphine framework (MOPF) directly. The MOPF employs riboflavin sodium phosphate (C17H20N4NaO9P) as an organic ligand as well as a nitrogen and phosphorus source to conjugate with Ni(NO3)2 · 6H2O. As an anode for LIBs, PCS was demonstrated with discharge capacities that were able to reach 386.5 mAh · g-1 after 100 cycles at a current density of 0.05 A · g-1. Besides, the stable reversible capacities were obtained from ∼459 mAh · g-1 to ∼91.8 mAh · g-1 when the current density was varied from 0.05 to 1 A · g-1. The good anode performance is attributed to the unique structure of PCS and (N, Ni, P) tri-doping which introduces the additional capacities due to the presence of the 'reservoir effect'. Moreover, the electrochemical analysis implied that the surface-limited capacitive behavior dominantly contributes to the lithium ion storage capacity of the PCS anode.

Near work, outdoor activity, and myopia in children in rural China: the Handan offspring myopia study.

BACKGROUND: The near work and outdoor activity are the most important environmental risk factors for myopia. However, data from Chinese rural children are relatively rare and remain controversial. Therefore, the purpose of this study was to evaluate the relationship of both near work and outdoor activities with refractive error in rural children in China. METHODS: In this cross-sectional study, 572 (65.1%) of 878 children (6-18 years of age) were included from the Handan Offspring Myopia Study (HOMS). Information from the parents on these children, as well as the parent's non-cycloplegic refraction, were obtained from the database of the Handan Eye Study conducted in the years 2006-2007. A comprehensive vision examination, including cycloplegic refraction, and a related questionnaire, were assessed on all children. RESULTS: The overall time spent on near work and outdoor activity in the children was 4.8 ± 1.6 and 2.9 ± 1.4 h per day, respectively. Myopic children spent more time on near work (5.0 ± 1.7 h vs.4.7 ± 1.6 h, p = 0.049), while no significant difference was found in outdoor activity hours (2.8 ± 1.3 h vs. 3.0 ± 1.4 h, p = 0.38), as compared to non-myopic children. In the multiple logistic analysis, in general, no association between near work and myopia was found after adjusting for the children's age, gender, parental refractive error, parental educational level, and daily outdoor activity hours [odds ratio (OR), 95% confidence interval (CI): 1.10, 0.94-1.27]. However, a weak protective effect of the outdoor activity on myopia was found (OR, 95% CI: 0.82, 0.70-0.96), after adjusting for similar confounders. CONCLUSIONS: In general, no association between near work and myopia was found, except for the high near work subgroup with moderate outdoor activity levels. A weak protective effect of outdoor activity on myopia in Chinese rural children was observed.

Eye exercises of acupoints: their impact on myopia and visual symptoms in Chinese rural children.

BACKGROUND: Chinese traditional "eye exercises of acupoints" have been advocated as a compulsory measure to reduce visual symptoms, as well as to retard the development of refractive error, among Chinese students for decades. The exercises are comprised of a 5-min, bilateral eye acupoint self-massage. This study evaluated the possible effect of these eye exercises among Chinese rural students. METHODS: Eight hundred thirty-six students (437 males, 52.3 %), aged 10.6 ± 2.5 (range 6-17) years from the Handan Offspring Myopia Study (HOMS) who completed the eye exercises and vision questionnaire, the convergence insufficiency symptom survey (CISS) questionnaire, and had a cycloplegic refraction were included in this study. RESULTS: 121 (14.5 %) students (64 males, 52.9 %) performed the eye exercises of acupoints in school. The multiple odds ratio (OR) and 95 % confidence interval (CI) for those having a "serious attitude" towards performing the eye exercises (0.12, 0.03-0.49) demonstrated a protective effect for myopia, after adjusting for the children's age, gender, average parental refractive error, and the time spent on near work and outdoor activity. The more frequently, and the more seriously, the students performed the eye exercises each week, the less likely was their chance of being myopic (OR, 95 % CI: 0.17, 0.03-0.99), after adjusting for the same confounders. However, neither the "seriousness of attitude" of performing the eye exercises (multiple ß coefficients: -1.58, p = 0.23), nor other related aspects of these eye exercises, were found to be associated with the CISS score in this sample. CONCLUSIONS: The traditional eye exercises of acupoints appeared to have a modest protective effect on myopia among these Chinese rural students aged 6-17 years. However, no association between the eye exercises and near vision symptoms was found.

Near work, outdoor activity, and their association with refractive error.

PURPOSE: To assess the relationship between near work, outdoor activity, and refractive error in schoolchildren in Beijing. METHODS: The Beijing Myopia Progression Study is a hospital-based myopia study, in which 386 students from primary (aged 6 to 12 years) and secondary (aged 13 to 17 years) schools in the inner city of Beijing were enrolled. Cycloplegic refraction and a detailed questionnaire probing near, intermediate, and distance visual activities were completed. RESULTS: Three hundred seventy (95.9%) of 386 students with complete cycloplegic autorefraction and myopia questionnaire data were enrolled in this study. Children with more near work time did not exhibit a significantly more myopic refraction in both the primary and secondary school levels after adjusting for the children's gender, outdoor activity time, and average parental refractive error. A significant association between outdoor activity time (in hours per day) and the children's spherical equivalent (in diopters) was found in the primary school students (ß = 0.27, p = 0.03) but not in the secondary school students (ß = 0.04, p = 0.70) after adjusting for similar confounders. The time spent on outdoor sports and outdoor leisure in the primary school students was also significantly associated with the children's spherical equivalent (ß = 0.46, p = 0.04 and ß = 0.31, p = 0.02, respectively). Primary school students with more time outdoors exhibited relatively less myopic refraction than their peers (ptrend = 0.0003), but this relation was not demonstrated in the secondary school children (ptrend = 0.53) after adjusting for similar confounders. CONCLUSIONS: Higher levels of outdoor activity were associated with less myopic refraction in primary school students in the inner city of Beijing. Near work activity was not found to be associated with refraction at either school level.

Characterization of Adeno-Associated Virus Capsid Proteins by Microflow Liquid Chromatography Coupled with Mass Spectrometry.

Adeno-associated virus (AAV) has been widely used to treat various human diseases as an important delivery vector for gene therapy due to its low immunogenicity and safety. AAV capsids proteins are comprised of three capsid viral proteins (VP; VP1, VP2, VP3). The capsid proteins play a key role in viral vector infectivity and transduction efficiency. To ensure the safety and efficacy of AAV gene therapy products, the quality of AAV vector capsid proteins during development and production should be carefully monitored and controlled. Microflow liquid chromatography coupled with mass spectrometry provides superior sensitivity and fast analysis capability. It showed significant advantages in the analysis of low- concentration and large numbers of AAV samples. The intact mass of capsid protein can be accurately determined using high-resolution mass spectrometry (MS). And MS also provides highly confident confirmation of sequence coverage and post-translational modifications site identification and quantitation. In this study, we used microflow liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the characterization of AAV2 capsid protein. we obtained nearly 100% sequence coverage of low-concentration AAV2 capsid protein (8 × 1011 GC/mL). More than 30 post-translational modifications (PTMs) sites were identified, the PTMs types included deamidation, oxidation and acetylation. From this study, the proposed microflow LC-MS/MS method provides a sensitive and high throughput approach in the characterization of AAVs and other biological products with low abundance.

Eye exercises of acupoints: their impact on refractive error and visual symptoms in Chinese urban children.

BACKGROUND: Traditional Chinese eye exercises of acupoints involve acupoint self-massage. These have been advocated as a compulsory measure to reduce ocular fatigue, as well as to retard the development of myopia, among Chinese school children. This study evaluated the impact of these eye exercises among Chinese urban children. METHODS: 409 children (195 males, 47.7%), aged 11.1 ± 3.2 (range 6-17) years, from the Beijing Myopia Progression Study (BMPS) were recruited. All had completed the eye exercise questionnaire, the convergence insufficiency symptom survey (CISS), and a cycloplegic autorefraction. Among these, 395 (96.6%) performed the eye exercises of acupoints. Multiple logistic regressions for myopia and multiple linear regressions for the CISS score (after adjusting for age, gender, average parental refractive error, and time spent doing near work and outdoor activity) for the different items of the eye exercises questionnaire were performed. RESULTS: Only the univariate odds ratio (95% confidence interval) for "seriousness of attitude" towards performing the eye exercises of acupoints (0.51, 0.33-0.78) showed a protective effect towards myopia. However, none of the odds ratios were significant after adjusting for the confounding factors. The univariate and multiple ß coefficients for the CISS score were -2.47 (p = 0.002) and -1.65 (p = 0.039), -3.57 (p = 0.002) and -2.35 (p = 0.042), and -2.40 (p = 0.003) and -2.29 (p = 0.004), for attitude, speed of exercise, and acquaintance with acupoints, respectively, which were all significant. CONCLUSIONS: The traditional Chinese eye exercises of acupoints appeared to have a modest effect on relieving near vision symptoms among Chinese urban children aged 6 to 17 years. However, no remarkable effect on reducing myopia was observed.

Emerging electro-driven technologies for phosphorus enrichment and recovery from wastewater: A review.

The recovery of phosphorus from wastewater is a critical step in addressing the scarcity of phosphorus resources. Electro-driven technologies for phosphorus enrichment have gathered significant attention due to their inherent advantages, such as mild operating conditions, absence of secondary pollution, and potential integration with other technologies. This study presents a comprehensive review of recent advancements in the field of phosphorus enrichment, with a specific focus on capacitive deionization and electrodialysis technologies. It highlights the underlying principles and effectiveness of electro-driven techniques for phosphorus enrichment while systematically comparing energy consumption, enrichment rate, and concentration factor among different technologies. Furthermore, the study provides a thorough analysis of the capacity of various technologies to selectively enrich phosphorus and proposes several methods and strategies to enhance selectivity. These insights offer valuable guidance for advancing the future development of electrochemical techniques with enhanced efficiency and effectiveness in phosphorus enrichment from wastewater.

Intraocular Pressure, Age, and Central Corneal Thickness in a Healthy Chinese Children Population: The Handan Offspring Myopia Study.

PURPOSE: To assess the distribution and factors related to intraocular pressure (IOP) in a Chinese children population in Northern China. METHODS: 1,238 offspring aged 6 to 18 years, residing in six villages, were recruited for this current study, the Handan Offspring Myopia Study. Participants underwent an interviewer-administered questionnaire and a complete ocular examination, including standardized measurement of IOP with Perkins applanation tonometry. RESULTS: 1,648 eyes of 828 children were included in the analysis. The mean IOP was 13.9 ± 1.6 mmHg. The mean IOP for the study population increased from 13.0 ± 1.5 mmHg for those 6 to 7 years of age to 14.2 ± 1.4 mmHg for those 15 years of age or older (P < .001). The mean central corneal thickness (CCT) was 548.7 ± 32.1 µm, and had no difference among different age groups and gender. In univariate regression analysis, age (0.12 mmHg per 1 year old), height (0.09 mmHg per 5 cm), weight (0.02 mmHg per kg), body mass index (0.07 mmHg per 1 m/kg2), systolic blood pressure (0.06 mmHg per 5 mmHg), CCT (0.06 mmHg per 5 µm), and SE (-0.11 mmHg per 1 D) were correlated with IOP. In multivariate regression analysis, higher IOP was only associated with older ages (p = .002) and thicker CCT (p = .001). CONCLUSION: The mean IOP in healthy rural Chinese children aged 6-18 years is about 14 mmHg, which is lower than in adults in the same locality. The mean IOP is slightly increasing with age during childhood, which is opposite to the result among adults. Age and CCT are the major independent factors associated with IOP.

Flow-electrode capacitive deionization: A review and new perspectives.

Flow-electrode capacitive deionization (FCDI), as a novel electro-driven desalination technology, has attracted growing exploration towards brackish water treatment, hypersaline water treatment, and selective resource recovery in recent years. As a flow-electrode-based electrochemical technology, FCDI has similarities with several other electrochemical technologies such as electrochemical flow capacitors and semi-solid fuel cells, whose performance are closely coupled with the characteristics of the flow-electrodes. In this review, we sort out the potentially parallel mechanisms of electrosorption and electrodialysis in the FCDI desalination process, and make clear the importance of the flowable capacitive electrodes. We then adopt an equivalent circuit model to distinguish the resistances to ion transport and electron transport within the electrodes, and clarify the importance of electronic conductivity on the system performance based on a series of electrochemical tests. Furthermore, we discuss the effects of electrode selection and flow circulation patterns on system performance (energy consumption, salt removal rate), review the current treatment targets and system performance, and then provide an outlook on the research directions in the field to support further applications of FCDI.

Separation, identification and quantification of associated impurities in cobratide using sheathless CE-MS and CE-UV.

Cobratide is a peptide drug extracted from the venom of Chinese cobra, and has been widely used in the clinical treatment of chronic, intractable and persistent pain. In a recent study, it was reported that it has the potential to treat COVID-19. In order to control the quality of commercial cobratide drugs, a protocol was established for the separation, identification and quantification of cobratide and its associated impurities, in which sheathless capillary electrophoresis-mass spectrometry (CE-MS) was used for identification and a rapid capillary electrophoresis-ultraviolet-visible detector (CE-UV) method was developed for accurate quantification. Separation conditions that affect the resolution and MS intensities of cobratide and its associated impurities were investigated, including pH value, concentration of background electrolyte (BGE), ratio of organic additive and sample solution. The optimized CE conditions (BGE: 50 mM NH4Ac, pH 4.0; sample solution: deionized water) were used for both sheathless CE-MS and CE-UV methods. Three associated impurities were separated and identified for the first time by sheathless CE-MS. Then, a rapid CE-UV method was validated and used for accurate quantification of cobratide and its associated impurities. The CE-UV method showed good linearity between concentration and corrected peak area of cobratide in the concentration range of 5.36-536.30 µg mL-1. The limit of quantification of the CE-UV method was 4.16 µg mL-1. The relative standard deviations of migration time were less than 1% for both intra-day and inter-day experiments, and those of corrected peak area were less than 5%. Finally, different cobratide drugs were analyzed to evaluate the batch-to-batch consistency. This established protocol combining sheathless CE-MS and CE-UV methods would provide useful information for both quality control and process analysis of peptide drugs.

[Immunohistochemical expression of HO-1 in traumatic human brain tissue].

OBJECTIVE: To investigate the expression of heme oxygenase-1 (HO-1) at different intervals and to provide evidence for estimation on injury intervals after brain contusion in human. METHODS: Twenty-four patients died of serious brain injury were assigned as injury group and 4 patients died of non-brain injury were served as control group. HO-1 expression was analyzed in brain tissue at different time intervals (3 h, 6-9 h, 12-24 h, 36 h-3d, 5-8d, 17-20d) by immunohistochemistry and auto-image analysis system. RESULTS: The level of HO-1 expression started to increase in 3 h after brain contusion compared to the control group (P < 0.05). The level of HO-1 expression highest level in 12-24 h group, and maintained high level in 36 h-3 d, then decreased gradually. CONCLUSION: The expression of HO-1 might be a strong evidence for human brain contusion time estimation.

Effects of Dietary Supplementation with High Fiber (Stevia Residue) on the Fecal Flora of Pregnant Sows.

This study aimed to investigate the effects of different supplementation levels of stevia residues in high-fiber diets on the fecal microorganisms of pregnant sows. Forty-eight first-farrowing Danish Landrace sows with similar weight, age, and due date were randomly divided into four groups. The control group was fed a normal diet (CON; 3.15% crude fiber content), and the treatment groups were supplemented with 20% stevia residue (SRL), 30% stevia residue (SRM), or 40% stevia residue (SRH) stevia residue, with crude fiber content of 7.79%, 9.15%, and 10.68%, respectively. The test period was 135 days, and Illumina Miseq high-throughput sequencing was used to test the fecal bacteria of the pregnant sows on day 56. Compared to the control group, species diversity was greater in the 30% stevia residue group. At the phylum level, Firmicutes and Spirochaetes had the greatest relative abundances; Firmicutes was most abundant in the SRM group, and Spirochaetes was most abundant in the CON group. At the genus level, Lactobacillus, Streptococcus, and Lachnospiraceae_XPB1014_group, and Treponema_2 had the highest relative abundances in the stevia-residue treatments. Among them, Lactobacillus and Treponema_2 were more abundant in SRL, Streptococcus was more abundant in SRH, and Lachnospiraceae_XPB1014_group was more abundant in SRM. Linear discriminant analysis effect size (LEfSe) showed that the bacterial species differed among the stevia residue treatments. SRL mainly included g__Lactobacillus and g__Romboutsi, SRM mainly included g__Lachnospiraceae_XPB1014_group and g__Ruminococcaceae_UCG_014, SRH mainly included g__Streptococcus, and CON mainly included g__Treponema_2, f__Clostridiaceae_1, g__norank_f__Muribaculaceae, and g__norank_f__p_251_o5. Stevia-residue supplementation significantly increased the abundance of beneficial bacteria, such as g__Lachnospiraceae_XPB1014_group, g__Christensenellaceae_R-7_group, and g__Ruminococcaceae_UCG-005 (p < 0.05), and significantly reduced the abundance of harmful bacteria, such as Treponema_2 (p < 0.05). Moreover, compared with the control group, the stevia-residues treatment groups reduced the feed cost 8.33%, 12.50%, and 13.33% per sow per day, respectively. In conclusion, high-fiber diets can improve the intestinal balance and evenness index of the intestinal flora in pregnant sows, promote the relative abundance of beneficial bacteria, reduce the relative abundance of harmful bacteria, and reduce feed cost. The optimal supplementation level of the stevia residue was found to be 30%.

[Determination of A2 ß-casein and total ß-casein in cow milk and milk powder by capillary zone electrophoresis].

ß-casein (ß-CN) is one of the major casein proteins in cow milk. There are 13 different variants documented for ß-CN in cow milk, among which A1 and A2 are the major variants. The separation and quantitation of A2 ß-CN are imperative for A2 dairy products. A new capillary zone electrophoresis (CZE) method with UV detection at 214 nm was established for the separation and quantification of the A2 variant and total ß-CN content in cow milk and milk powders. The separation of ß-CN variants was achieved on bare fused silica capillaries (50 µm×30/40 cm (effective/total length)). The separation buffer was a mixture of 4 mol/L urea, 0.2% (mass fraction) hydroxypropyl methylcellulose, 140 mmol/L citric acid, and 50 mmol/L disodium hydrogen phosphate buffer (pH 2.7). The corrected peak areas and the concentrations of total ß-CN and the A2 variant showed good linearity, with correlation coefficients (r2) ranging from 0.9968 to 0.9997. The intra-day precisions for A2 ß-CN and total ß-CN determination in four samples (two pasteurized milk samples and two milk powder samples) were in the ranges of 2.4%-4.7% and 2.6%-4.8%, respectively. The inter-day precisions for A2 ß-CN and total ß-CN determination in four samples were in the ranges of 4.0%-6.3% and 3.9%-6.7%, respectively. The recoveries of A2 and total ß-CN ranged from 85.5% to 106.4%. With the established CZE method, the A2 ß-CN variant and total ß-CN protein in liquid and powder bovine milk products could be separated and accurately quantified. By calculating the A2 ß-CN content in the total ß-CN, the quality of A2 dairy products can be evaluated, and this in turn would aid in the protection of consumer rights.

Rapid Determination of Full and Empty Adeno-Associated Virus Capsid Ratio by Capillary Isoelectric Focusing.

Adeno-associated virus (AAV) is one of the most promising gene transfer vector types featuring long-term gene expression and low toxicity. The lack of pathogenicity and the availability of many serotypes augmented the applicability of AAV virions in gene therapy applications. The recombinant AAV capsid includes the therapeutic protein-coding transgene as well as a promoter to initiate translation and a poly A sequence portion for stabilization. Current AAV manufacturing technologies, however, cannot guarantee the generation of only full capsids, i.e., including the entire required genome. Partially filled and empty capsids are also part of the product, decreasing in this way the efficacy and safety upon clinical translation. Therefore, rapid, accurate and QC friendly analysis of the full and empty capsid ratio is of high importance during AAV vector manufacturing and release testing. In this paper, an automated capillary isoelectric focusing technique is introduced, readily applicable in the biopharmaceutical industry for fast and efficient determination of the full and empty capsid ratio. The method also reveals information about the proportion of partially filled capsids. For higher resolution (<0.1 pI unit), mixtures of wide and narrow range ampholytes were utilized. The isoelectric point and peak area percentage reproducibility (RSD) of the mixed ampholyte assay were as low as 1.67% and 2.45 %, respectively, requiring only 65 nL of sample volume per injection.