Scalable encapsulation of hepatocytes by electrostatic spraying

Encapsulating cells by polyelectrolyte complex coacervation can be accomplished at physiological temperature and buffer conditions. One of the oppositely charged polyelectrolytes in the microcapsule core can be collagen or any other natural extra-cellular matrices suitable for cellular support while the other polyelectrolyte forms the ultra-thin shell to ensure efficient mass transfer. These microcapsules with ultra-thin shell are difficult to produce in large quantities due to their fragility. In this study, electrostatic spraying technique was used to achieve a scalable production of one such type of microcapsules formed by complex coacervation between the cationic methylated collagen and anionic terpolymer of hydroxylethyl methacrylate, methyl methacrylate and methylacrylic acid (HEMA-MMA-MAA). It was found that the microcapsule sizes were dependent on several important operational parameters, such as the diameter of the spraying needle, the flow rate of the hepatocytes-collagen mixture and the voltage of the electrical field. The microcapsules with diameters of 200-800 microm and a narrow size distribution (standard deviation of 5-28%) were successfully produced. The above parameters also influenced the hepatocyte viability and functions. With a practical encapsulation rate of up to 55 ml/h per orifice required in bio-artificial liver-assisted device applications, we have produced large quantities of microcapsules maintaining comparable cell viability (>87%), mechanical stability and bio-functions to the manually extruded microcapsules.     

Construction and characterization of a live attenuated Shigella flexneri 2a vaccine strain, sf301 Delta virG and dsbA33G]

Construction and characterization of a live attenuated Shigella flexneria 2a sf301 vaccine strain to prevent the endemic of shigellosis. Using Chinese majority epidemic Shigella flexneri 2a serotype sf301 as the target, p Delta virG, a deletion derivation of the virG gene in the SacB suicide vector pCVD442 and pDsbA33G, an mutant of a disulfide bond catalyst DsbA, replaced its 33 amiano acid Cystine by Glycerin in pCVD442, were used to generate a attenuated mutant strain sf301: Delta virG: DsbA33 G. Its virulence was evaluated by Sereny test, the invasive ability was detected by HeLa cell invasive assay, immunogenicity was detected by immunized Guinea pigs through inoculated guinea pigs' conjunctive sac. Sereny test was negative and HeLa invasive assay showed sf301: Delta virG: DsbA33 G retained partial invasive ability. In contrast to control group, sf301: Delta virG: DsbA33 G could induced significantly high antibody levels of IgA and IgG against sf301 LPS in animal's mucosal lavage fluids and sera in both primary immunization protocol and boosting protocol. The numbers of ASCs in local draining lymph nodes and spleens were significantly higher than control group. The immune response to sf301: Delta virG: DsbA33 G could provide completely protection from the challenge of wild type sf301. sf301: Delta virG: DsbA33 G is a safe and effective oral candidate vaccine to prevent the infection of Shigella strains.     

Role of dual-site phospholamban phosphorylation in intermittent hypoxia-induced cardioprotection against ischemia-reperfusion injury

Cardioprotection by intermittent high-altitude (IHA) hypoxia against ischemia-reperfusion (I/R) injury is associated with Ca(2+) overload reduction. Phospholamban (PLB) phosphorylation relieves cardiac sarcoplasmic reticulum (SR) Ca(2+)-pump ATPase, a critical regulator in intracellular Ca(2+) cycling, from inhibition. To test the hypothesis that IHA hypoxia increases PLB phosphorylation and that such an effect plays a role in cardioprotection, we compared the time-dependent changes in the PLB phosphorylation at Ser(16) (PKA site) and Thr(17) (CaMKII site) in perfused normoxic rat hearts with those in IHA hypoxic rat hearts submitted to 30-min ischemia (I30) followed by 30-min reperfusion (R30). IHA hypoxia improved postischemic contractile recovery, reduced the maximum extent of ischemic contracture, and attenuated I/R-induced depression in Ca(2+)-pump ATPase activity. Although the PLB protein levels remained constant during I/R in both groups, Ser(16) phosphorylation increased at I30 and 1 min of reperfusion (R1) but decreased at R30 in normoxic hearts. IHA hypoxia upregulated the increase further at I30 and R1. Thr(17) phosphorylation decreased at I30, R1, and R30 in normoxic hearts, but IHA hypoxia attenuated the depression at R1 and R30. Moreover, PKA inhibitor H89 abolished IHA hypoxia-induced increase in Ser(16) phosphorylation, Ca(2+)-pump ATPase activity, and the recovery of cardiac performance after ischemia. CaMKII inhibitor KN-93 also abolished the beneficial effects of IHA hypoxia on Thr(17) phosphorylation, Ca(2+)-pump ATPase activity, and the postischemic contractile recovery. These findings indicate that IHA hypoxia mitigates I/R-induced depression in SR Ca(2+)-pump ATPase activity by upregulating dual-site PLB phosphorylation, which may consequently contribute to IHA hypoxia-induced cardioprotection against I/R injury.     

Effect of light quality on leaf photosynthetic characteristics and fruit quality of peach (<Emphasis Type=Italic>Prunus persica</Emphasis> L. Batch)

Different light filters affect leaf photosynthetic features and fruit quality. Consequently, selecting the appropriate covering filter for rain-shelter cultivation of peaches is a key part of successful production. We used a late-maturing peach variety ‘Xiahui 8’ to study differences in leaf photosynthetic features, chlorophyll fluorescence characteristics, and fruit quality under neutral, red, yellow, green, and blue filter, with natural light as control. The results showed that the leaf photosynthetic ability and internal quality under the neutral filter treatment were elevated compared with the control, and the appearance color was the same as the control. Leaves under neutral filter could maintain higher photosynthetic ability than other filter treatments. In addition, the fruits could also keep higher quality when treated with neutral filter. Therefore, the application of neutral filter in rain-shelter cultivation of ‘Xinhui 8’ peaches is recommended for maintaining high photosynthetic capacity and for improving fruit quality.     

An ignored and potential source of taste and odor (T&O) issues—biofilms in drinking water distribution system (DWDS)

It is important for water utilities to provide esthetically acceptable drinking water to the public, because our consumers always initially judge the quality of the tap water by its color, taste, and odor (T&O). Microorganisms in drinking water contribute largely to T&O production and drinking water distribution systems (DWDS) are known to harbor biofilms and microorganisms in bulk water, even in the presence of a disinfectant. These microbes include T&O-causing bacteria, fungi, and algae, which may lead to unwanted effects on the organoleptic quality of distributed water. Importantly, the understanding of types of these microbes and their T&O compound-producing mechanisms is needed to prevent T&O formation during drinking water distribution. Additionally, new disinfection strategies and operation methods of DWDS are also needed for better control of T&O problems in drinking water. This review covers: (1) the microbial species which can produce T&O compounds in DWDS; (2) typical T&O compounds in DWDS and their formation mechanisms by microorganisms; (3) several common factors in DWDS which can influence the growth and T&O generation of microbes; and (4) several strategies to control biofilm and T&O compound formation in DWDS. At the end of this review, recommendations were given based on the conclusion of this review.

     

Differential expression of microRNAs in TM3 Leydig cells of mice treated with brain‐derived neurotrophic factor

Brain‐derived neurotrophic factor (BDNF) is a neurotrophin that can promote the development and proliferation of neurons. BDNF has been found to be involved in male reproduction. Leydig cells in testicular interstitial tissues can secrete testosterone in a luteinizing hormone‐dependent manner. We showed that BDNF and its receptor TrkB were expressed in mice TM3 Leydig cells in the present study. Furthermore, BDNF can promote proliferation of mouse TM3 Leydig cells in vitro. Results of microRNA (miRNA) deep sequencing showed that BDNF can alter the expression profile of miRNAs in TM3 Leydig cells. Eighty‐three miRNAs were significantly different in the BDNF‐treated and control groups (fold change of >2.0 or <0.5, P < 0.05) wherein 40 were upregulated and 43 were downregulated. The expression levels of miR‐125a‐5p, miR‐22‐5p, miR‐342‐59, miR‐451a, miR‐148a‐5p, miR‐29b‐3p, miR‐199b‐5p, and miR‐145a‐5p were further confirmed by quantitative real‐time polymerase chain reaction. Bioinformatic analysis revealed that miRNAs regulated a large number of genes with different functions. Pathway analysis indicated that miRNAs participate in the pathways involved in signal transduction, cancer, metabolism, endocrine system, immune system, and nerve system. This study indicated that miRNAs might be involved in the BDNF‐regulated cellular functions of Leydig cells.     

The Role of Heat Shock Protein 90B1 in Patients with Polycystic Ovary Syndrome

Polycystic ovary syndrome (PCOS) is a heterogenetic disorder in women that is characterized by arrested follicular growth and anovulatory infertility. The altered protein expression levels in the ovarian tissues reflect the molecular defects in folliculogenesis. To identify aberrant protein expression in PCOS, we analyzed protein expression profiles in the ovarian tissues of patients with PCOS. We identified a total of 18 protein spots that were differentially expressed in PCOS compared with healthy ovarian samples. A total of 13 proteins were upregulated and 5 proteins were downregulated. The expression levels of heat shock protein 90B1 (HSP90B1) and calcium signaling activator calmodulin 1 (CALM1) were increased by at least two-fold. The expression levels of HSP90B1 and CALM1 were positively associated with ovarian cell survival and negatively associated with caspase-3 activation and apoptosis. Knock-down of HSP90B1 with siRNA attenuated ovarian cell survival and increased apoptosis. In contrast, ovarian cell survival was improved and cell apoptosis was decreased in cells over-expressing HSP90B1. These results demonstrated the pivotal role of HSP90B1 in the proliferation and survival of ovarian cells, suggesting a critical role for HSP90B1 in the pathogenesis of PCOS. We also observed a downregulation of anti-inflammatory activity-related annexin A6 (ANXA6) and tropomyosin 2 (TPM2) compared with the normal controls, which could affect cell division and folliculogenesis in PCOS. This is the first study to identify novel altered gene expression in the ovarian tissues of patients with PCOS. These findings may have significant implications for future diagnostic and treatment strategies for PCOS using molecular interventions.     

Effects of branched‐chain amino acid supplementation on fecundity,lifespan and flight ability of Helicoverpa armigera

The effects on cotton bollworm, Helicoverpa armigera, of supplementing the benchmark larval feeding formula with branched‐chain amino acids (BCAAs) were analyzed. The results show that supplementary BCAAs can increase adult longevity, and are conducive to improving development and reproductive capacity. In particular, by supplementing the ration with the three proteinogenic BCAAs at concentrations ten times that of the benchmark formula, all the selected physiological indices of cotton bollworm health were improved. The larval duration, pupae weight, pupation rate, fecundity and adult longevity for specimens fed the supplemented formula were 15.8 days, 0.348 g, 100%, 1340.7 eggs and 27.3 days, respectively. The same indicators for larvae fed the benchmark formula were 16.6 days, 0.306 g, 97.9%, 1167.0 eggs and 12.0 days, respectively. This study provides a reference for future research on amino acid supplementation and increasing lifespans