Investigation of adverse reactions in healthcare personnel working in Level 3 barrier protection PPE to treat COVID-19.

PURPOSE OF THE STUDY: The aim of our study was to investigate potential adverse reactions in healthcare professionals working in Level 3 barrier protection personal protective equipment (L3PPE) to treat patients with COVID-19. STUDY DESIGN: By using a convenience sampling approach, 129 out of 205 randomly selected healthcare professionals from the First Affiliated Hospital of Zhejiang University School of Medicine were invited to take part in a WeChat messaging app survey, Questionnaire Star, via a survey link. Healthcare personnel details were collected, including profession, years of professional experience and adverse reactions while wearing L3PPE. Survey results were divided by profession and years of professional experience; differences in adverse reactions were compared. RESULTS: Among the 129 healthcare professionals surveyed, 21 (16.28%) were doctors and 108 (83.72%) were nurses. A total of 122 (94.57%) healthcare professionals experienced discomfort while wearing L3PPE to treat patients with COVID-19. The main reasons for adverse reactions and discomfort include varying degrees of adverse skin reactions, respiratory difficulties, heat stress, dizziness and nausea. Doctors had a lower incidence of rashes (χ2=4.519, p=0.034) and dizziness (χ2=4.123, p=0.042) when compared with nurses. Junior (8.5 years of experience or fewer) healthcare personnel also experienced a higher rate of heat stress when compared with senior personnel (more than 8.5 years greater) (χ2=5.228, p=0.022). CONCLUSION: More attention should be offered to healthcare personnel wearing L3PPE to treat patients with COVID-19 because they are susceptible to developing adverse reactions.

[Micronutrients intake of pregnant women from 8 cities of China during 2011 to 2012].

OBJECTIVE: To investigate micronutrients intake of pregnant women in 8 cities. METHODS: The multi-stage stratified cluster sampling method was applied in 8 cities( Beijing, Shanghai, Guangzhou, Lanzhou, Shenyang, Chengdu, Zhengzhou and Suzhou) from September 2011 to March 2012 to recruit healthy pregnant women, and then general information and dietary status were collected through face to face investigation. Nutrients intakes were calculated and analyzed then. RESULTS: There were 479 pregnantwomen in analysis, 58. 9% of whom were 25. 1 to 30 years old and the number of pregnant women in every trimester was approximately equal, counting for 1/3 respectively. Principal components analysis showed pregnant women in Lanzhou share poorest status of micronutrients. For pregnant women in Lanzhou, intakes of riboflavin, vitamin C, vitamin A, folacin and calcium were lowest compared with pregnant women from other 7 cities. The most B-vitamins intakes of pregnant women in Zhengzhou, and vitamin A, vitamin C, calcium intake of pregnant women in Shanghai were highest, respectively. CONCLUSION: Of all the participants, nutritional status of pregnant women in Lanzhou was poorest compare with those in other cities and more attention should be paid on their dietary nutrition. Compared with result of 2002, nutrients intakes of pregnant women were improved in this survey.

[Interference of Lumican Regulates the Invasion and Migration of Liver Cancer Cells].

OBJECTIVE: To explore the effect of silencing lumican on the invasion and migration of liver cancer cells. METHODS: Lumican was silenced by shRNA in liver cancer cells (HepG2 and MHCC97H). The mRNA levels of lumican were detected by qRT-PCR. Cell invasion was measured by Transwell. Cell migration was tested via wound healing. The protein levels of lumican,MMP-9,VEGF,ERK1,JNK,p-ERK1 and p-JNK were measured by Western blot. RESULTS: Liver cancer cells (HepG2 and MHCC97H) had higher levels of mRNA and protein of lumican compared with normal hepatocyte L02 (P<0.01). shRNA lowered the levels of mRNA and protein of lumican (P<0.01),and weakened the invasion and migration of cancer cells (P<0.01). The expressions of MMP-9 and VEGF decreased with the shRNA silence (P<0.01). shRNA also reduced the protein level of p-ERK1 and p-JNK (P<0.01). CONCLUSION: Silencing lumican by shRNA attenuates the invasion and migration of liver cancer cells via inhibiting the activation of ERK1/JNK pathway.

Expression of a constitutively active nitrate reductase variant in tobacco reduces tobacco-specific nitrosamine accumulation in cured leaves and cigarette smoke.

Burley tobaccos (Nicotiana tabacum) display a nitrogen-use-deficiency phenotype that is associated with the accumulation of high levels of nitrate within the leaf, a trait correlated with production of a class of compounds referred to as tobacco-specific nitrosamines (TSNAs). Two TSNA species, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N-nitrosonornicotine (NNN), have been shown to be strong carcinogens in numerous animal studies. We investigated the potential of molecular genetic strategies to lower nitrate levels in burley tobaccos by overexpressing genes encoding key enzymes of the nitrogen-assimilation pathway. Of the various constructs tested, only the expression of a constitutively active nitrate reductase (NR) dramatically decreased free nitrate levels in the leaves. Field-grown tobacco plants expressing this NR variant exhibited greatly reduced levels of TSNAs in both cured leaves and mainstream smoke of cigarettes made from these materials. Decreasing leaf nitrate levels via expression of a constitutively active NR enzyme represents an exceptionally promising means for reducing the production of NNN and NNK, two of the most well-documented animal carcinogens found in tobacco products.

Thermal demands and its interactions with environmental factors account for national-level variation in aggression.

Literature shows that psychological phenomena, including values (e.g., individualism vs. collectivism), personality, and behaviors (e.g., prosocial and aggressive behavior), are geographically clustered. The effects of temperature on interpersonal and intergroup aggression have been studied by many social psychologists. To date the interactions between temperature and other geographical factors have not been addressed. This study is aiming to examine the effects of thermal demands and the moderating effects of natural geographical factors on aggressive behavior at national level. Data for 156 societies was obtained from publicly available databases. Consistent with the life-history theory, results of this study showed that aggressive behavior has a positive relationship with heat demands, and a negative relationship with cold demands. Aggressive behavior is also positively correlated with sunlight and altitude, and negatively correlated with coastline vicinity. Forest, coastline vicinity, and rainfall moderated the effect of thermal demands on aggressive behavior. In societies with more forests, with more rainfall, and closer to coastline, the negative effects of cold demands on aggressive behavior are stronger.

The Prosocial Outgrowth of Filial Beliefs in Different Cultures: A Conditional Mediation Model Analysis.

Filial piety is a concept originated from ancient China which contains norms of children's feelings, attitudes, and behaviors toward their parents. The dual filial piety model (DFPM) differentiated two types of filial belief: reciprocal vs. authoritarian filial piety (RFP vs. AFP). Recent scholars suggest that the functions of filial piety may differ across cultures. This study examined the mediating effects of empathy, moral identity, gratitude, and sense of indebtedness in the relationship between filial piety and prosocial behavior (PB) and the moderating effects of nation. Questionnaires measuring filial piety, PB, moral identity, gratitude, and sense of indebtedness were administrated to Chinese and Indonesian participants. Moderated mediation modeling was conducted to analyze data. The results showed that empathy, moral identity, gratitude, and a sense of indebtedness have significant mediating effects in the association of filial piety and PB. And nation served as a moderator. (1) RFP could promote PB via enhanced empathy, moral identity, gratitude, and a sense of indebtedness, both among Chinese and Indonesian participants, while AFP did the same job only among Indonesian participants. (2) Among Chinese participants, AFP was not directly associated with PB, but was negatively associated with PB via reduced gratitude and a sense of indebtedness. (3) Nation (China vs. Indonesia) moderated the direct or indirect effect of RFP/AFP on PB, with RFP exerting stronger positive effects on outcome variables among Chinese (relative to Indonesian) participants and AFP exerting stronger positive effects on outcome variables among Indonesian (relative to Chinese) participants. These results showed that RFP can promote prosocial development by the cultivation of empathy, moral identity, gratitude, and a sense of indebtedness, regardless of whether the participants grew up in China or other cultural backgrounds. But the effect of AFP on PB was significantly conditioned by culture. This suggests that the function of RFP may be a cultural universal. However, the mechanisms that AFP influences PB can differ considerably across cultures. Findings of this study further indicate that filial piety beliefs may facilitate prosocial development in the ways conditioned by cultures.

Reduced JKAP correlates with advanced disease features, inflammation, as well as increased exacerbation risk and severity in asthmatic children.

BACKGROUND: This study aimed to investigate the correlation of JNK pathway-associated phosphatase (JKAP) with clinical features, inflammation, exacerbation risk, and severity in asthmatic children. METHODS: Asthmatic exacerbation children (N = 90), asthmatic remission children (N = 90), and healthy controls (N = 90) were enrolled in this case-control study, whose venous blood samples were collected after enrollment for routine blood test, JKAP, and inflammatory cytokines detection by enzyme-linked immune sorbent assay. The clinical features included demographic data, family history of asthma, and pulmonary ventilation function. RESULTS: JKAP level was the lowest in asthmatic exacerbation children, followed by asthmatic remission children and healthy controls. ROC curve revealed good ability of JKAP in distinguishing three groups from each other, especially in telling asthmatic exacerbation children from healthy controls (AUC: 0.926; 95%CI: 0.887-0.965). In addition, JKAP was negatively correlated with eosinophil count, immunoglobulin E (IgE), tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1ß), interleukin-6 (IL-6), and interleukin-17 (IL-17), positively correlated with forced expiratory volume in 1 sec/forced vital capacity (FEV1/FVC) and FEV1 (%predicted) in asthmatic exacerbation children. Whereas in asthmatic remission children, JKAP was negatively correlated with eosinophil count, TNF-α, IL-1ß, IL-6, and IL-17 and positively correlated with FEV1 (%predicted), but not with IgE or FEV1/FVC. In healthy controls, the correlation of JKAP with clinical features and inflammatory cytokines was non-obvious. For exacerbation severity, JKAP was the highest in mild exacerbation children, followed by moderate exacerbation children, and severe exacerbation children. CONCLUSION: JKAP serves as a potential biomarker for asthmatic susceptibility, inflammation, exacerbation risk, and severity in children.

Constitutive activation of nitrate reductase in tobacco alters flowering time and plant biomass.

Pyridine alkaloids produced in tobacco can react with nitrosating agents such as nitrite to form tobacco-specific nitrosamines (TSNA), which are among the most notable toxicants present in tobacco smoke. The market type known as burley tobacco is particularly susceptible to TSNA formation because its corresponding cultivars exhibit a nitrogen-use-deficiency phenotype which results in high accumulation of nitrate, which, in turn, is converted to nitrite by leaf surface microbes. We have previously shown that expression of a constitutively activated nitrate reductase (NR) enzyme dramatically decreases leaf nitrate levels in burley tobacco, resulting in substantial TSNA reductions without altering the alkaloid profile. Here, we show that plants expressing a constitutively active NR construct, designated 35S:S523D-NR, display an early-flowering phenotype that is also associated with a substantial reduction in plant biomass. We hypothesized that crossing 35S:S523D-NR tobaccos with burley cultivars that flower later than normal would help mitigate the undesirable early-flowering/reduced-biomass traits while maintaining the desirable low-nitrate/TSNA phenotype. To test this, 35S:S523D-NR plants were crossed with two late-flowering cultivars, NC 775 and NC 645WZ. In both cases, the plant biomass at harvest was restored to levels similar to those in the original cultivar used for transformation while the low-nitrate/TSNA trait was maintained. Interestingly, the mechanism by which yield was restored differed markedly between the two crosses. Biomass restoration in F1 hybrids using NC 645WZ as a parent was associated with delayed flowering, as originally hypothesized. Unexpectedly, however, crosses with NC 775 displayed enhanced biomass despite maintaining the early-flowering trait of the 35S:S523D-NR parent.

Folding zones inside the ribosomal exit tunnel.

Helicity of membrane proteins can be manifested inside the ribosome tunnel, but the determinants of compact structure formation inside the tunnel are largely unexplored. Using an extended nascent peptide as a molecular tape measure of the ribosomal tunnel, we have previously demonstrated helix formation inside the tunnel. Here, we introduce a series of consecutive polyalanines into different regions of the tape measure to monitor the formation of compact structure in the nascent peptide. We find that the formation of compact structure of the polyalanine sequence depends on its location. Calculation of free energies for the equilibria between folded and unfolded nascent peptides in different regions of the tunnel shows that there are zones of secondary structure formation inside the ribosomal exit tunnel. These zones may have an active role in nascent-chain compaction.

Less mechanical loading attenuates osteoarthritis by reducing cartilage degeneration, subchondral bone remodelling, secondary inflammation, and activation of NLRP3 inflammasome.

AIMS: Osteoarthritis (OA) is a disabling joint disorder and mechanical loading is an important pathogenesis. This study aims to investigate the benefits of less mechanical loading created by intermittent tail suspension for knee OA. METHODS: A post-traumatic OA model was established in 20 rats (12 weeks old, male). Ten rats were treated with less mechanical loading through intermittent tail suspension, while another ten rats were treated with normal mechanical loading. Cartilage damage was determined by gross appearance, Safranin O/Fast Green staining, and immunohistochemistry examinations. Subchondral bone changes were analyzed by micro-CT and tartrate-resistant acid phosphatase (TRAP) staining, and serum inflammatory cytokines were evaluated by enzyme-linked immunosorbent assay (ELISA). RESULTS: Our radiographs showed that joint space was significantly enlarged in rats with less mechanical loading. Moreover, cartilage destruction was attenuated in the less mechanical loading group with lower histological damage scores, and lower expression of a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-5, matrix metalloproteinase (MMP)-3, and MMP-13. In addition, subchondral bone abnormal changes were ameliorated in OA rats with less mechanical loading, as reduced bone mineral density (BMD), bone volume/tissue volume (BV/TV), and number of osteophytes and osteoclasts in the subchondral bone were observed. Finally, the level of serum inflammatory cytokines was significantly downregulated in the less mechanical loading group compared with the normal mechanical loading group, as well as the expression of NACHT, LRR, and PYD domains-containing protein 3 (NLRP3), caspase-1, and interleukin 1ß (IL-1ß) in the cartilage. CONCLUSION: Less mechanical loading alleviates cartilage destruction, subchondral bone changes, and secondary inflammation in OA joints. This study provides fundamental insights into the benefit of non-weight loading rest for patients with OA. Cite this article: Bone Joint Res 2020;9(10):731-741.

Mapping the electrostatic potential within the ribosomal exit tunnel.

Electrostatic potentials influence interactions among proteins and nucleic acids, the orientation of dipoles and quadrupoles, and the distribution of mobile charges. Consequently, electrostatic potentials can modulate macromolecular folding and conformational stability, as well as rates of catalysis and substrate binding. The ribosomal exit tunnel, along with its resident nascent peptide, is no less susceptible to these consequences. Yet, the electrostatics inside the tunnel have never been measured. Here we map both the electrostatic potential and accessibilities along the length of the tunnel and determine the electrostatic consequences of introducing a charged amino acid into the nascent peptide. To do this we developed novel probes and strategies. Our findings provide new insights regarding the dielectric of the tunnel and the dynamics of its local electric fields.

Regional discrimination and propagation of local rearrangements along the ribosomal exit tunnel.

All proteins, from bacteria to man, are made in the ribosome and are elongated, one residue at a time, at the peptidyl transferase center. This growing peptide chain wends its way through the ribosomal tunnel to the exit port, ~100Å from the peptidyl transferase center. We have identified locations in the tunnel that sense and respond to single side chains of the nascent peptide to induce local conformational changes. Moreover, side-chain sterics and rearrangements deep in the tunnel influence the disposition of residues 45Å away at the exit port and are consistent with side-chain-induced axial retraction of the peptide backbone. These coupled responses are neither haphazard nor uniform along the tunnel. Rather, they are confined to discriminating zones in the tunnel and are sequence specific. Such discerning communication may contribute to folding events and mechanisms governing sequence-specific signaling between different regions of the tunnel during translation.

Nascent peptide side chains induce rearrangements in distinct locations of the ribosomal tunnel.

Although we have numerous structures of ribosomes, none disclose side-chain rearrangements of the nascent peptide during chain elongation. This study reports for the first time that rearrangement of the peptide and/or tunnel occurs in distinct regions of the tunnel and is directed by the unique primary sequence of each nascent peptide. In the tunnel mid-region, the accessibility of an introduced cysteine to a series of novel hydrophilic maleimide reagents increases with increasing volume of the adjacent chain residue, a sensitivity not manifest at the constriction and exit port. This surprising result reveals molecular movements not yet resolvable from structural studies. These findings map solvent-accessible volumes along the tunnel and provide novel insights critical to our understanding of allosteric communication within the ribosomal tunnel, translational arrest, chaperone interaction, folding, and rates of elongation.

Electrostatics in the ribosomal tunnel modulate chain elongation rates.

Electrostatic potentials along the ribosomal exit tunnel are nonuniform and negative. The significance of electrostatics in the tunnel remains relatively uninvestigated, yet they are likely to play a role in translation and secondary folding of nascent peptides. To probe the role of nascent peptide charges in ribosome function, we used a molecular tape measure that was engineered to contain different numbers of charged amino acids localized to known regions of the tunnel and measured chain elongation rates. Positively charged arginine or lysine sequences produce transient arrest (pausing) before the nascent peptide is fully elongated. The rate of conversion from transiently arrested to full-length nascent peptide is faster for peptides containing neutral or negatively charged residues than for those containing positively charged residues. We provide experimental evidence that extraribosomal mechanisms do not account for this charge-specific pausing. We conclude that pausing is due to charge-specific interactions between the tunnel and the nascent peptide.

Activity of the 5' regulatory regions of the rice polyubiquitin rubi3 gene in transgenic rice plants as analyzed by both GUS and GFP reporter genes.

Ubiquitin is an abundant protein involved in protein degradation and cell cycle control in plants and rubi3 is a polyubiquitin gene isolated from rice (Oryza sativa L.). Using both GFP and GUS as reporter genes, we analyzed the expression pattern of the rubi3 promoter as well as the effects of the rubi3 5'-UTR (5' untranslated region) intron and the 5' terminal 27 bp of the rubi3 coding sequence on the activity of the promoter in transgenic rice plants. The rubi3 promoter with the 5'-UTR intron was active in all the tissue and cell types examined and supported more constitutive expression of reporter genes than the maize Ubi-1 promoter. The rubi3 5'-UTR intron mediated enhancement on the activity of its promoter in a tissue-specific manner but did not alter its overall expression pattern. The enhancement was particularly intense in roots, pollen grains, inner tissue of ovaries, and embryos and aleurone layers in maturing seeds. The translational fusion of the first 27 bp of the rubi3 coding sequence to GUS gene further enhanced GUS expression directed by the rubi3 promoter in all the tissues examined. The rubi3 promoter should be an important addition to the arsenal of strong and constitutive promoters for monocot transformation and biotechnology.

Transcriptional and post-transcriptional enhancement of gene expression by the 5' UTR intron of rice rubi3 gene in transgenic rice cells.

Introns play a very important role in regulating gene expression in eukaryotes. In plants, many introns enhance gene expression, and the effect of intron-mediated enhancement (IME) of gene expression is reportedly often more profound in monocots than in dicots. To further gain insight of IME in monocot plants, we quantitatively dissected the effect of the 5' UTR intron of the rice rubi3 gene at various gene expression levels in stably transformed suspension cell lines. The intron enhanced the GUS reporter gene activity in these lines by about 29-fold. Nuclear run-on experiments demonstrated a nearly twofold enhancement by the 5' UTR intron at the transcriptional level. RNA analysis by RealTime quantitative RT-PCR assays indicated the intron enhanced the steady state RNA level of the GUS reporter gene by nearly 20-fold, implying a strong role of the intron in RNA processing and/or export. The results also implicated a moderate role of the intron in enhancement at the translational level ( approximately 45%). Moreover, results from a transient assay experiment using a shortened exon 1 sequence revealed an important role of exon 1 of rubi3 in gene expression. It may also hint a divergence in IME mechanisms between plant and animal cells. These results demonstrated transcriptional enhancement by a plant intron, but suggested that post-transcriptional event(s) be the major source of IME.

Gene expression enhancement mediated by the 5' UTR intron of the rice rubi3 gene varied remarkably among tissues in transgenic rice plants.

Introns are important sequence elements that modulate the expression of genes. Using the GUS reporter gene driven by the promoter of the rice (Oryza sativa L.) polyubiquitin rubi3 gene, we investigated the effects of the 5' UTR intron of the rubi3 gene and the 5' terminal 27 bp of the rubi3 coding sequence on gene expression in stably transformed rice plants. While the intron enhanced GUS gene expression, the 27-bp fused to the GUS coding sequence further augmented GUS expression level, with both varying among different tissues. The intron elevated GUS gene expression mainly at mRNA accumulation level, but also stimulated enhancement at translational level. The enhancement on mRNA accumulation, as determined by realtime quantitative RT-PCR, varied remarkably with tissue type. The augmentation by the intron at translational level also differed by tissue type, but to a lesser extent. On the other hand, the 27-bp fusion further boosted GUS protein yield without affecting mRNA accumulation level, indicating stimulation at translation level, which was also affected by tissue type. The research revealed substantial variation in the magnitudes of intron-mediated enhancement of gene expression (IME) among tissues in rice plants and the importance of using transgenic plants for IME studies.

Expression of the bacteriophage T4 lysozyme gene in tall fescue confers resistance to gray leaf spot and brown patch diseases.

Tall fescue (Festuca arundinacea Schreb.) is an important turf and forage grass species worldwide. Fungal diseases present a major limitation in the maintenance of tall fescue lawns, landscapes, and forage fields. Two severe fungal diseases of tall fescue are brown patch, caused by Rhizoctonia solani, and gray leaf spot, caused by Magnaporthe grisea. These diseases are often major problems of other turfgrass species as well. In efforts to obtain tall fescue plants resistant to these diseases, we introduced the bacteriophage T4 lysozyme gene into tall fescue through Agrobacterium-mediated genetic transformation. In replicated experiments under controlled environments conducive to disease development, 6 of 13 transgenic events showed high resistance to inoculation of a mixture of two M. grisea isolates from tall fescue. Three of these six resistant plants also displayed significant resistance to an R. solani isolate from tall fescue. Thus, we have demonstrated that the bacteriophage T4 lysozyme gene confers resistance to both gray leaf spot and brown patch diseases in transgenic tall fescue plants. The gene may have wide applications in engineered fungal disease resistance in various crops.

Secondary structure formation of a transmembrane segment in Kv channels.

Transmembrane segments in the intact voltage-gated potassium (Kv) channel are helical. To ascertain whether this helicity could first be manifested inside the ribosomal tunnel, we generated biogenic peptide intermediates of Kv1.3 and mass-tagged the cysteine-scanned S6 transmembrane segment using pegylation (PEG-MAL) and calmodulation (CaM-MAL). For reference, we created an extended peptide that was used as a "molecular tape measure" of the ribosomal tunnel and determined that the functional length of the tunnel is 99-112 A. We demonstrate that the S6 segment forms a compact structure inside the ribosomal tunnel and that the N-terminal half of S6 compacts more than the C-terminal half of S6. These results bear on the earliest folding events during biogenesis of ion channels.