Glycosylation of H4 influenza strains with pandemic potential and susceptibilities to lung surfactant SP-D.

We recently reported that members of group 1 influenza A virus (IAV) containing H2, H5, H6, and H11 hemagglutinins (HAs) are resistant to lung surfactant protein D (SP-D). H3 viruses, members of group 2 IAV, have high affinity for SP-D, which depends on the presence of high-mannose glycans at glycosite N165 on the head of HA. The low affinity of SP-D for the group 1 viruses is due to the presence of complex glycans at an analogous glycosite on the head of HA, and replacement with high-mannose glycan at this site evoked strong interaction with SP-D. Thus, if members of group 1 IAV were to make the zoonotic leap to humans, the pathogenicity of such strains could be problematic since SP-D, as a first-line innate immunity factor in respiratory tissues, could be ineffective as demonstrated in vitro. Here, we extend these studies to group 2 H4 viruses that are representative of those with specificity for avian or swine sialyl receptors, i.e., those with receptor-binding sites with either Q226 and G228 for avian or recent Q226L and G228S mutations that facilitate swine receptor specificity. The latter have increased pathogenicity potential in humans due to a switch from avian sialylα2,3 to sialylα2,6 glycan receptor preference. A better understanding of the potential action of SP-D against these strains will provide important information regarding the pandemic risk of such strains. Our glycomics and in vitro analyses of four H4 HAs reveal SP-D-favorable glycosylation patterns. Therefore, susceptibilities to this first-line innate immunity defense respiratory surfactant against such H4 viruses are high and align with H3 HA glycosylation.

Preparation and characterization of a solid dispersion of Hexahydrocolupulone and its application in the preservation of fresh apple juice.

Hops extracts and their derivatives have many important biological activities, among them, excellent antibacterial and antioxidant properties make them a promising food preservative. However, poor water solubility limits their application in the food industry. This work aimed to improve the solubility of Hexahydrocolupulone (HHCL) by preparing solid dispersion (SD) and investigating the application of the obtained products (HHCL-SD) in actual food systems. HHCL-SD was prepared by solvent evaporation with PVPK30 as a carrier. The solubility of HHCL was dramatically increased to 24.72 mg/mL（25 ℃)by preparing HHCL-SD, much higher than that of raw HHCL (0.002 mg/mL). The structure of HHCL-SD and the interaction between HHCL and PVPK30 were analyzed. HHCL-SD was confirmed to have excellent antibacterial and antioxidant activities. Furthermore, the addition of HHCL-SD proved to be beneficial for the sensory, nutritional quality, and microbiological safety of fresh apple juice, hence prolonging its shelf-life.

A novel sucrose-inducible expression system and its application for production of biomass-degrading enzymes in Aspergillus niger.

BACKGROUND: Filamentous fungi are extensively exploited as important enzyme producers due to the superior secretory capability. However, the complexity of their secretomes greatly impairs the titer and purity of heterologous enzymes. Meanwhile, high-efficient evaluation and production of bulk enzymes, such as biomass-degrading enzymes, necessitate constructing powerful expression systems for bio-refinery applications. RESULTS: A novel sucrose-inducible expression system based on the host strain Aspergillus niger ATCC 20611 and the ß-fructofuranosidase promoter (PfopA) was constructed. A. niger ATCC 20611 preferentially utilized sucrose for rapid growth and ß-fructofuranosidase production. Its secretory background was relatively clean because ß-fructofuranosidase, the key enzyme responsible for sucrose utilization, was essentially not secreted into the medium and the extracellular protease activity was low. Furthermore, the PfopA promoter showed a sucrose concentration-dependent induction pattern and was not subject to glucose repression. Moreover, the strength of PfopA was 7.68-fold higher than that of the commonly used glyceraldehyde-3-phosphate dehydrogenase promoter (PgpdA) with enhanced green fluorescence protein (EGFP) as a reporter. Thus, A. niger ATCC 20611 coupled with the PfopA promoter was used as an expression system to express a ß-glucosidase gene (bgla) from A. niger C112, allowing the production of ß-glucosidase at a titer of 17.84 U/mL. The crude ß-glucosidase preparation could remarkably improve glucose yield in the saccharification of pretreated corncob residues when added to the cellulase mixture of Trichoderma reesei QM9414. The efficacy of this expression system was further demonstrated by co-expressing the T. reesei-derived chitinase Chi46 and ß-N-acetylglucosaminidase Nag1 to obtain an efficient chitin-degrading enzyme cocktail, which could achieve the production of N-acetyl-D-glucosamine from colloidal chitin with a conversion ratio of 91.83%. Besides, the purity of the above-secreted biomass-degrading enzymes in the crude culture supernatant was over 86%. CONCLUSIONS: This PfopA-driven expression system expands the genetic toolbox of A. niger and broadens the application field of the traditional fructo-oligosaccharides-producing strain A. niger ATCC 20611, advancing it to become a high-performing enzyme-producing cell factory. In particular, the sucrose-inducible expression system possessed the capacity to produce biomass-degrading enzymes at a high level and evade endogenous protein interference, providing a potential purification-free enzyme production platform for bio-refinery applications.

Aerobic granulation and microbial community succession in sequencing batch reactors treating the low strength wastewater: The dual effects of weak magnetic field and exogenous signal molecule.

The application of magneto-biological effects in wastewater treatment has been brought under the spotlight recently. This work explored the dual effects of magnetic field (MF) and exogenous N-hexanoyl-l-homoserine lactone (C6-HSL) on activated sludge granulation. Results showed that exposure to MF and C6-HSL obviously accelerated the aerobic granulation process and promoted the secretion of extracellular polymeric substances, especially polysaccharides, humic acid-like substances, aromatic proteins, and tryptophan-like substrates. Illumina MiSeq sequencing results indicated that the introduction of MF and C6-HSL can increase the diversity and richness of microbial community without antagonism, and the biological basis for rapid granulation process in this study was the enrichment of slow-growing bacteria Candidatus_Competibacter. Besides, the overgrowth of filamentous bacteria Thiothrix could be suppressed due to the presence of MF, improving the stabilities of aerobic granular sludge. This study provides a new understanding of the MF and C6-HSL effects on rapid aerobic granulation when treating the low-strength wastewater.

Aromatherapy Plus Music Therapy Improve Pain Intensity and Anxiety Scores in Patients With Breast Cancer During Perioperative Periods: A Randomized Controlled Trial.

INTRODUCTION: To investigate the effect of perioperative aromatherapy (AT) or/plus music therapy (MT) on pain and anxiety level, and the potential mechanism in women experiencing breast cancer surgery. METHODS: A total of 160 patients with breast cancer were randomly assigned in a 1:1:1:1 ratio to receive usual care (UC), AT, MT, or combination therapy (CT) during perioperative periods. Pain intensity and anxiety scores were measured by visual analog scale. Interleukin (IL)-6 and high mobility group box 1 (HMGB-1) were measured by enzyme-linked immunosorbent assay. RESULTS: There was a significant group effect on pain, anxiety, and IL-6 and HMGB-1 levels, with the greatest improvement occurring in the CT group (P < .001). Compared with the UC group, the AT and MT groups had lower mean changes of pain intensity and IL-6 and HMGB-1 levels, and greater anxiety reduction (P < .001). However, the differences between the AT and MT groups did not reach significance (P > .05). CONCLUSION: In patients with breast cancer, perioperative CT achieves greater pain improvement and anxiety reduction and less marked increase in IL-6 and HMGB-1 levels. These results suggest that CT is an acceptable complementary and alternative medicine for breast cancer patients.

Structural basis of malarial parasite RIFIN-mediated immune escape against LAIR1.

Malaria infection by Plasmodium falciparum continues to pose a global threat to the human population. P. falciparum expresses variable erythrocyte surface antigens such as RIFINs. Public antibodies with LAIR1 insertion have been identified from malarial patients against a subset of RIFINs. In this study, we solve a LAIR1-binding RIFIN structure: the complex structures of two RIFINs bound to mutated or wild-type LAIR1 in two distinct patterns. Notably, the two RIFINs engage similar binding sites on LAIR1 with different angles, and the RIFIN-binding sites overlap with the collagen-binding site. Surprisingly, RIFINs use completely different binding sites to bind to LAIR1 or LILRB1, indicating the kaleidoscopic change of RIFINs. We then verify that RIFIN could induce LAIR1-mediated cell signaling, and LAIR1-containing antibodies could block the pathway. The findings of this study provide structural insights into the mechanism of the immune escape of P. falciparum and the endless arms race between parasite and host.

[Effect of root development on the root and periodontal absorptions of maxillary impacted anterior teeth after orthodontic traction].

PURPOSE: To investigate the effect of root development on root and periodontal tissues of maxillary impacted anterior teeth after orthodontic traction based on cone-beam CT (CBCT). METHODS: Thirty-four patients (34 teeth) requiring orthodontic traction for maxillary impacted anterior teeth in Southwest Medical University Affiliated Stomatological Hospital from July 2018 to March 2019 were selected. The patients were divided into incomplete development group and complete development group according to root development, each with 17 cases. All patients received orthodontic traction treatment. Root length, thickness of root canal wall, pulp vitality, width of the attachment ridge, bone mineral density around root and orthodontic traction time were compared between the two groups. Statistic analysis was completed by SPSS 20.0 software package. RESULTS: Root length of complete development group was significantly longer than that of incomplete development group before treatment(P<0.05). After treatment, the root length of incomplete development group was significantly longer than that of the complete development group(P<0.05). The thickness of root canal wall was significantly greater in complete development group than in incomplete development group before treatment (P<0.05), which had no significant difference between groups after treatment(P>0.05). The pulp vitality and width of the attachment ridge had no significant difference between two groups after treatment(P>0.05). Bone mineral density around root was significantly decreased in both groups after treatment(P<0.05), and was lower in complete development group than in incomplete development group(P<0.05). The orthodontic traction time in the complete development group was significantly longer than that in incomplete development group(P<0.05). CONCLUSIONS: In orthodontic traction treatment, the impacted anterior teeth with undeveloped roots have advantages of mild root absorption, rapid periodontal tissue reconstruction, short orthodontic traction time with the root, thickness of root canal wall and pulp vitality in the normal range.

Demands of experiential training for ICU nurses in Hunan of China.

OBJECTIVE: To assess intensive care unit (ICU) nurses' demands for specialized experiential training and to provide inputs for developing an experiential training program for ICU nurses. METHODS: A questionnaire for assessing ICU nurses' demands for experiential specialty training was distributed to 360 ICU nurses, selected through purposive sampling from two secondary hospitals and six tertiary hospitals in Hunan Province, China. RESULTS: Of the survey participants, 63.6% had undergone a specialty training program for ICU nurses. Of these individuals, 53.0% were satisfied with the training. Certification as a qualified nurse was considered an essential criterion for admission of trainees into the program by 81.8% of respondents, while 77.1% of respondents considered clinical working experience to be a critical requirement for selecting trainers. A total of 48.1% of the respondents preferred part-time training, and 36.1% considered a training cycle of 9-12 weeks to be reasonable. Moreover, they felt that the training methods should be tailored to different stages of the experiential learning cycle. Demands for experiential training among ICU nurses were quantified, with high demand reflected in an overall score of 4.41 ± 0.48. The "intensive care technology" experiential training module was ranked highest in terms of demand, with the top five sub-modules being specialty operating technology (4.67 ± 0.53), care of critically ill patients (4.66 ± 0.55), critical patient rescue procedures (4.65 ± 0.56), assessing monitoring indexes (4.63 ± 0.56), and the application of relevant instruments (4.61 ± 0.57). CONCLUSION: Nearly half of the respondents indicated that their experiences of specialty training programs were not satisfactory, and they had high demands for experiential training. Thus, to optimize training outcomes, continuous updating of training methods is essential. Moreover, a systematic, comprehensive, and multilevel experiential training program that targets the specific needs of ICU nurses is essential.

Insights into functional microbial succession during nitrogen transformation in an ectopic fermentation system.

The ectopic fermentation system (EFS) is an advanced technology for treating farm wastewater, and it reduces ammonia nitrogen emission and nitrogen loss of fermentation products. This study observed the functional bacteria succession related to nitrogen metabolism in EFS by high throughput sequencing, and evaluated their associations with environmental factors. Results revealed that with the changes of temperature, pH, moisture content, and nitrogen content during fermentation, the species richness and diversity of ammonia oxidizing bacteria (AOB) with amoA increased, but those of denitrifying bacteria carrying nirK and nosZ decreased. During the fermentation process, the dominant bacterial populations of AOB and denitrifying bacteria changed significantly, and different bacterial populations showed different positive/negative correlations with the environmental factors. This study revealed the role of functional bacteria in ammonia removal and nitrogen conservation of EFS, and provided a theoretical basis for the improvement of microbial agents and EFS application.

[Effects of aroma therapy and music intervention on pain and anxious for breast cancer patients in the perioperative period].

OBJECTIVE: To investigate the effect of the aroma therapy and music intervention on anxious and pain for the breast cancer patients in the perioperative period and the potential mechanisms.
 Methods: A total of 100 breast cancer patients who received surgical treatment in the comprehensive hospitals of Hunan province were recruited for this study. Patients were assigned randomly into a control group, an aroma therapy group, a music intervention group, and a joint-therapy group (n=25 per group). The patients in the control group received regular post-surgical nursery, while the patients from other groups received aroma therapy, music intervention, or both in addition to the regular nursery. The scale of anxiety and pain were measured. The measurements were carried at three time points, namely 30 min before the surgery (T1), 30 min after the recovery period of anesthesia (T2), and 4 hours after the removal of anesthesia tubing (T3). Repeated ANOVA was used to perform statistic analysis.
 Results: The scale of pain was significantly increased at the post-operation (T2, T3) compared to pre-surgery (T1). The therapeutic group showed significant decrease in pain at post-operation (T3) comparing with the control group (P<0.05). The scale of anxiety was the highest at pre-surgery (T1). During anaesthesia recovery, the anxiety of patients at post-operation T2 and T3 in the therapeutic groups significantly decreased compared with the control group (P<0.05).
 Conclusion: Both the aroma therapy and the music therapy can decrease the stress-responsive anxiety and pain for the breast cancer patients in the perioperative period.

Recombinant Chimpanzee Adenovirus Vaccine AdC7-M/E Protects against Zika Virus Infection and Testis Damage.

The recent outbreak of Zika virus (ZIKV) has emerged as a global health concern. ZIKV can persist in human semen and be transmitted by sexual contact, as well as by mosquitoes, as seen for classical arboviruses. We along with others have previously demonstrated that ZIKV infection leads to testis damage and infertility in mouse models. So far, no prophylactics or therapeutics are available; therefore, vaccine development is urgently demanded. Recombinant chimpanzee adenovirus has been explored as the preferred vaccine vector for many pathogens due to the low preexisting immunity against the vector among the human population. Here, we developed a ZIKV vaccine based on recombinant chimpanzee adenovirus type 7 (AdC7) expressing ZIKV M/E glycoproteins. A single vaccination of AdC7-M/E was sufficient to elicit potent neutralizing antibodies and protective immunity against ZIKV in both immunocompetent and immunodeficient mice. Moreover, vaccinated mice rapidly developed neutralizing antibody with high titers within 1 week postvaccination, and the elicited antiserum could cross-neutralize heterologous ZIKV strains. Additionally, ZIKV M- and E-specific T cell responses were robustly induced by AdC7-M/E. Moreover, one-dose inoculation of AdC7-M/E conferred mouse sterilizing immunity to eliminate viremia and viral burden in tissues against ZIKV challenge. Further investigations showed that vaccination with AdC7-M/E completely protected against ZIKV-induced testicular damage. These data demonstrate that AdC7-M/E is highly effective and represents a promising vaccine candidate for ZIKV control.IMPORTANCE Zika virus (ZIKV) is a pathogenic flavivirus that causes severe clinical consequences, including congenital malformations in fetuses and Guillain-Barré syndrome in adults. Vaccine development is a high priority for ZIKV control. In this study, to avoid preexisting anti-vector immunity in humans, a rare serotype chimpanzee adenovirus (AdC7) expressing the ZIKV M/E glycoproteins was used for ZIKV vaccine development. Impressively, AdC7-M/E exhibited exceptional performance as a ZIKV vaccine, as follows: (i) protective efficacy by a single vaccination, (ii) rapid development of a robust humoral response, (iii) durable immune responses, (iv) robust T cell responses, and (v) sterilizing immunity achieved by a single vaccination. These advantages of AdC7-M/E strongly support its potential application as a promising ZIKV vaccine in the clinic.

A molasses habitat-derived fungus Aspergillus tubingensis XG21 with high ß-fructofuranosidase activity and its potential use for fructooligosaccharides production.

The industrial microorganisms used for fructooligosaccharides (FOS) synthesis are generally fermented with sucrose as carbon source to induce the production of ß-fructofuranosidase (FFase) having transfructosylation activity. Consequently, isolation of novel FFase producers from a sucrose-enriched biotope would help improve FOS productivity and reduce the process cost. Here, three fungi isolated from a unique sugarcane molasses habitat were found to possess FFase activity and one of them, XG21, exhibited a high capacity to synthesize FOS. Analysis of its morphological properties and ribosomal internal transcribed spacer (ITS) sequence allowed the taxonomic position to be assigned and it was thus identified as Aspergillus tubingensis XG21. It could utilize various potential carbon sources for vigorous growth, but only produced high-level FFase activity on sucrose. Furthermore, the transfructosylation ability and FOS synthesis were analyzed by TLC and HPLC. During the transfructosylation reaction, an increase in sucrose concentration led to the remarkable enhancement in FOS formation with the maximum content of up to 56.9% within 8 h. Finally, the sugarcane molasses was used to cultivate A. tubingensis XG21 and the optimal FFase activity reached up to 558.3 U/g, which was 88.9% higher than that with sucrose as carbon source. These results indicate that A. tubingensis XG21 can be considered as a new genetic resource adapted to cheaply available carbon sources for FOS production.

Enhancing fructooligosaccharides production by genetic improvement of the industrial fungus Aspergillus niger ATCC 20611.

Aspergillus niger ATCC20611 is one of the most potent filamentous fungi used commercially for production of fructooligosaccharides (FOS), which are prospective components of functional food by stimulating probiotic bacteria in the human gut. However, current strategies for improving FOS yield still rely on production process development. The genetic engineering approach hasn't been applied in industrial strains to increase FOS production level. Here, an optimized polyethylene glycol (PEG)-mediated protoplast transformation system was established in A. niger ATCC 20611 and used for further strain improvement. The pyrithiamine resistance gene (ptrA) was selected as a dominant marker and protoplasts were prepared with high concentration (up to 108g-1 wet weight mycelium) by using mixed cell wall-lysing enzymes. The transformation frequency with ptrA can reach 30-50 transformants per µg of DNA. In addition, the efficiency of co-transformation with the EGFP reporter gene (egfp) was high (approx. 82%). Furthermore, an activity-improved variant of ß-fructofuranosidase, FopA(A178P), was successfully overexpressed in A. niger ATCC 20611 by using the transformation system. The transformant, CM6, exhibited a 58% increase in specific ß-fructofuranosidase activity (up to 507U/g), compared to the parental strain (320U/g), and effectively reduced the time needed for completion of FOS synthesis. These results illustrate the feasibility of strain improvement through genetic engineering for further enhancement of FOS production level.

Orthodontic treatment in a periodontal patient with pathologic migration of anterior teeth.

A 22-year-old man with severe periodontitis and pathologic tooth migration sought orthodontic treatment. He was treated successfully with effective control of the strength and the direction of the orthodontic forces, and a combination of periodontic and orthodontic treatment. After 22 months of orthodontic treatment, his occlusal trauma resulting from pathologic tooth migration was relieved, a stable occlusion was achieved, and mutual aggravation of occlusal trauma and periodontitis was prevented. Furthermore, the patient's facial esthetics and self-confidence were improved. Newly formed trabecular bone could be seen in the periapical x-rays in some regions where resorption had been severe. This case report shows that resorption of alveolar bone can be prevented and that multidisciplinary orthodontic treatment of a periodontal patient with pathologic tooth migration is effective and helpful.