Development of an attenuated potato virus Y mutant carrying multiple mutations in helper-component protease for cross-protection.

Tobacco (Nicotiana tabacum) is one of the major cash crops in China. Potato virus Y (PVY), a representative member of the genus Potyvirus, greatly reduces the quality and yield of tobacco leaves by inducing veinal necrosis. Mild strain-mediated cross-protection is an attractive method of controlling diseases caused by PVY. Currently, there is a lack of effective and stable attenuated PVY mutants. Potyviral helper component-protease (HC-Pro) is a likely target for the development of mild strains. Our previous studies showed that the residues lysine at positions 124 and 182 (K124 and K182) in HC-Pro were involved in PVY virulence, and the conserved KITC motif in HC-Pro was involved in aphid transmission. In this study, to improve the stability of PVY mild strains, K at position 50 (K50) in KITC motif, K124, and K182 were separately substituted with glutamic acid (E), leucine (L), and arginine (R), resulting in a triple-mutant PVY-HCELR. The mutant PVY-HCELR had attenuated virulence and did not induce leaf veinal necrosis symptoms in tobacco plants and could not be transmitted by Myzus persicae. Furthermore, PVY-HCELR mutant was genetically stable after six serial passages, and only caused mild mosaic symptoms in tobacco plants even at 90 days post inoculation. The tobacco plants cross-protected by PVY-HCELR mutant showed high resistance to the wild-type PVY. This study showed that PVY-HCELR mutant was a promising mild mutant for cross-protection to control PVY.

Characterization of aroma-active compounds in Dongli by quantitative descriptive analysis, gas chromatography-triple quadrupole tandem mass spectrometry, and gas chromatography-olfactometry.

Dongli, or frozen pear, is a traditional Chinese snack with a unique flavor. This study identified the aroma-active volatile compounds (VOCs) in Dongli using quantitative descriptive analysis (QDA), gas chromatography-triple quadrupole tandem mass spectrometry (GC-MS/MS), and gas chromatography-olfactometry (GC-O). QDA indicated that Dongli of all cultivars presented increased sweet and wine aromas. A total of 21 VOCs were identified by GC-MS/MS. Bidirectional orthogonal partial least square (O2PLS) analysis, GC-O analysis, detection frequency analysis (DFA), and relative odor activity values (ROAV) showed that: estragole and anethole contributing "anise, green" aromas were the key aromatic VOCs of fresh pears, while ethyl butanoate, butyl acetate, heptyl acetate, benzaldehyde, and geranyl acetone contributing "sweet, fruity, green" aromas were the key aromatic VOCs of Dongli. The results revealed that the repeated freezing treatment promoted a unique aroma in pears. This study would contribute to developing new pear products. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-022-05463-8.

Multivariate analyses of the volatile components in fresh and dried turnip (Brassica rapa L.) chips via HS-SPME-GC-MS.

Turnip is a vegetable that has many health promoting effects. To diversify the usage and increase the consumption of turnip, the effects of hot air drying, infrared drying, explosion puff drying and freeze drying (FD) on the volatiles of turnip chips were studied. The volatiles of fresh turnip and dried turnip chips were isolated by HS-SPME-GC-MS and a total of 67 volatiles were identified. However, the volatiles in turnip chips dried by different methods are quite different. Based on principal component analysis and hierarchical cluster analysis, the volatiles of fresh turnip were distinguished from those of the dried chips and FD was separated from the other drying methods. As the result of orthogonal projection on latent structure-discriminant analysis (OPLS-DA), isothiocyanato-cyclopropane and (2-isothiocyanatoethyl)-benzene were identified as the characteristic volatiles of fresh turnip. While, 2-azido-2,3,3-trimethyl-butane and hexanal were identified as the characteristic volatiles for FD dried chips.

Exogenous p53 upregulated modulator of apoptosis (PUMA) decreases growth of lung cancer A549 cells.

PURPOSE: To investigate the influence of exogenous p53 upregulated modulator of apoptosis (PUMA) expression on cell proliferation and apoptosis in human non-small cell lung cancer A549 cells and transplanted tumor cell growth in nude mice. MATERIALS AND METHODS: A549 cells were divided into the following groups: control, non- carrier (NC), PUMA (transfected with pCEP4- (HA) 2-PUMA plasmid), DDP (10 µg/mL cisplatin treatment) and PUMA+DDP (transfected with pCEP4-(HA)2-PUMA plasmid and 10 µg/mL cisplatin treatment). The MTT method was used to detect the cell survival rate. Cell apoptosis rates were measured by flow cytometry, and PUMA, Bax and Bcl-2 protein expression levels were measured by Western blotting. RESULTS: Compared to the control group, the PUMA, DDP and PUMA+DDP groups all had significantly decreased A549 cell proliferation (p<0.01), with the largest reduction in the PUMA+DDP group. Conversely, the apoptosis rates of the three groups were significantly increased (P<0.01), and the PUMA and DDP treatments were synergistic. Moreover, Bax protein levels significantly increased (p<0.01), while Bcl-2 protein levels significantly decreased (p<0.01). Finally, both the volume and the weights of transplanted tumors were significantly reduced (p<0.01), and the inhibition ratio of the PUMA+DDP group was significantly higher than in the single DDP or PUMA groups. CONCLUSIONS: Exogenous PUMA effectively inhibited lung cancer A549 cell proliferation and transplanted tumor growth by increasing Bax protein levels and reducing Bcl-2 protein levels.

HO-1 induction by CO-RM2 attenuates TNF-α-induced cytosolic phospholipase A2 expression via inhibition of PKCα-dependent NADPH oxidase/ROS and NF-κB.

Rheumatoid arthritis (RA) is characterized by chronic inflammatory infiltration of the synovium and elevation of proinflammatory cytokines. Cytosolic phospholipase A2 (cPLA2) is involved in the development of inflammatory diseases. Heme oxygenase-1 (HO-1) has been shown to possess anti-inflammatory properties. The objective of the study was to investigate the detailed mechanisms of TNF-α-induced cPLA2 expression and to determine whether carbon monoxide releasing molecule-2 (CO-RM2) suppresses TNF-α-induced expression of NF-κB-related proinflammatory genes, including cPLA2, via HO-1 induction in RA synovial fibroblasts (RASFs). Here, we reported that TNF-α-induced cPLA2 expression was mediated through TNFR1/PKCα-dependent signaling pathways, including NADPH oxidase (NOX) activation/ROS production and NF-κB activation. CO-RM2 significantly suppressed TNF-α-induced cPLA2 expression by inhibiting the ROS generation and the phosphorylation of NF-κB p65 and IKK α/ß, but not the phosphorylation of p38 MAPK and JNK1/2. These results were further confirmed by a ChIP assay to detect the NF-κB DNA-binding activity. Our results demonstrated that induction of HO-1 by CO-RM2 exerted anti-inflammatory and antioxidant effects which were required in concert to prevent the activation of NF-κB leading to induction of various inflammatory genes implicated in the pathogenesis of RA.

[Effects of lac insect honeydew on the diversity of ground-dwelling ants in lac plantation].

By the method of pitfall trapping, an investigation was conducted on the diversity of ground-dwelling ants in a lac plantation in Yayi Town of Mojiang County, Yunnan Province of Southwest China in December 2009-May 2010, aimed to understand the effects of lac insect honeydew on the diversity of ground-dwelling ants. The presence or absence of lac insect honeydew and its dynamics all affected the species composition, abundance, and diversity of ground-dwelling ants. In the lac plantation with lac insect hosting, a total of 4953 ant individuals were collected, belonging to 34 species, 23 genera, and 5 subfamilies of Formicidae; whereas in the lac plantation without lac insect hosting, a total of 2416 ant individuals were collected, belonging to 30 species, 20 genera, and 5 subfamilies of Formicidae. The relative abundance, species richness (S), and ACE index in the lac plantation with lac insect hosting were higher than those in the lac plantation without lac insect hosting, and the common species and indicator species of ground-dwelling ants in the lac plantation with lac insect hosting differed from those in the lac plantation without lac insect hosting, suggesting that lac insect hosting altered the community structure of ground-dwelling ants. The adult lac insects excreted larger quantity of honeydew than the larval lac insects, and the relative abundance, S, and ACE index of ground-dwelling ants were higher at the adult stage than at the larval stage of lac insects. The common species and indicator species of ground-dwelling ants also had great differences between the two stages of lac insects.

[Monopolization of honeydew sources by Crematogaster macaoensis and its effects on lac production].

From October 2008 to May 2010, an investigation was made in a lac plantation in Yayi region of Mojiang County, Yunnan Province, China, aimed to understand the behavior of the monopolization of honeydew sources by ant Crematogaster macaoensis, and its effects on the sex ratio, mortality, fecundity, and lac production of Yunnan lac insect Kerria yunnanensis. The results showed that C. macaoensis fed and monopolized the honeydews around the clock during the whole life cycle of K. yunnanensis on the stick-lac, and the average number of visiting C. macaoensis ranged from 16.8 +/- 2.3 to 39.3 +/- 10.0 per 10 cm length of the stick-lac. C. macaoensis constructed shelters to prevent other animals from visiting the lac insect. C. macaoensis monopolization reduced the lac production of individual K. yunnanensi significantly, but had no significant effects on the lac production of whole lac insect colony. In addition, C. macaoensis monopolization reduced the mortality of K. yunnanensis significantly, and increased the percentage of K. yunnanensis females and the adult female fecundity. It was concluded that ant monopolization of the lac insect honeydew seemed to be beneficial to the lac production by the Yunnan lac insect.

[Sequence analysis of the phosphoprotein gene of peste des petits ruminants virus of Chinese origin].

The nucleotide sequences of P gene from a field strain of peste des petits ruminants virus (PPRV) ("China/Tib/Gej/07-30") was firstly determined. The P gene is 1,655 nucleotides long with two overlapping open reading frames (ORFs). The first ORF is 1530 nucleotides long and would produce P protein of 509 amino acid residues. The second ORF is 534 nucleotides long and would produce C protein of 177 amino acid residues. The first ORF produces a second mRNA transcript of 897 nucleotides long with an extra G nucleotide at position 751. Translation from this mRNA would produce V protein of 298 amino acid residues. The nucleotide and deduced amino acid sequence were compared with the homologous region of other PPRV isolates. At the amino acid level, the "China/Tib/Gej/07-30" shares homology of 86.10%-97.3%, 84.3%-94.9%, and 82.9%-96.3% for P, C, and V proteins respectively. Several sequence motifs in the P genes were identified on the basis of conservation in the PPRVs and the morbilliviruses.

[Sequence analysis of the matrix protein and fusion protein genes of a field peste des petits ruminants virus strain from Tibet, China].

The nucleotide sequences of M and F genes from a field strain of peste des petits ruminants virus (PPRV) ("China/Tib/Gej/07-30") was firstly determined. The M gene was 1 483 nucleotides in length with a single open reading frame (ORF), encoding a protein of 335 amino acids. The F gene was 2411 nucleotides in length, encoding a protein of 546 amino acids. The resulting nucleotide sequence and the deduced amino acid sequences were compared with the homologous regions of other PPRV isolates. The nucleotide sequences of M and F genes of the "China/Tib/Gej/07-30" was 92.4%-97.7% and 85.5%-96.1% identical to other PPRV isolates, respectively, while a homology of 97.0%-98.2% and 94.3%-98.2% could be observed at the amino acids level respectively. Several sequence motifs in the M and F genes had been identified on the basis of conservation in the PPRVs and the morbilliviruses. The 3' untranslated region of M gene was 443 nucleotides in length with 82.4%-93.5% identical to other PPRV isolates. The 5' untranslated region of F gene was 634 nucleotides in length with 76.2%-91.7% identical to other PPRV isolates.

[Coexistence mechanism of ant community in lac plantation under habitat heterogeneity].

In order to reveal the coexistence mechanism of ant community in lac plantation, an investigation was made on the ant community composition and the ability of ant species in discovering and holding food resources in a lac plantation in Yayi Town of Mojiang County, Yunnan Province, with the relationships between ant body size and its ability of finding food under habitat heterogeneity probed. There were six dominant ant species in the plantation, i. e., Tetraponera allaborans (Walker), Crematogaster macaoensis Wheeler, Crematogasterferrarii Emery, Dolichoderus thoracicus (Smith), Polyrhachis proxima Roger, and Camponotus parius Emery. The hind leg length (y) of the six ant species increased allometrically with their head width (x), and the regression equation was y = 0.56 + 1.02x + 5.97x2 - 10.85x3. Different ant species had significant differences in their actual and relative frequency in discovering food resources in different habitats, but habitat type had no significant effects on the actual frequency in holding food resources by the ant species. The ant species with bigger head width and bigger body size index could discover more food resources in simple habitat. In contrast, the ant species with smaller head width, shorter hind leg length, and smaller body size index could discover more food resources in complex habitat. The heterogeneity of habitat caused the coexistence of ants: the smaller ant species lived in complex habitat, while the larger ones lived in simple habitat. In addition, numerically dominant ant species were unable to possess all resources, and thereby, could provide the opportunity to other ant species for resources acquisition, making the species coexistence come true.

A study on the efficacy of body-mind-spirit group therapy for patients with breast cancer.

AIMS AND OBJECTIVES: This study aims to understand the effects of culturally enriched body-mind-spirit group therapy on anxiety, depression and holistic well-being among women with breast cancer and to examine patients' views on what aspects of group therapy worked to enhance their health. DESIGN: The study was designed using multiple methods, which consisted of a randomised controlled trial and a focus group interview. METHODS: A total of 16 subjects in the control group received the standard care of a physician's treatment at the outpatient department. In addition to standard care, 12 subjects in the experimental group received 10 sessions of weekly body-mind-spirit group therapy for 180 minutes each. This therapy integrates concepts and practices of traditional Chinese medicine and Western medicine (e.g. positive psychology and forgiveness therapy). The subjects in the experimental group were invited to participate in a focus group interview regarding their perceptions of the change mechanisms that occurred in group therapy. RESULTS: The results of analysis of covariance indicated that after a two-month trial, there was a similarity between the experimental and control groups in reducing the scores of Beck depression inventory and increasing the scores of body-mind-spirit well-being. However, subjects in the experimental group had a better reduction of the scores of state anxiety inventory than subjects in the control group. The qualitative analysis yielded eight domains: (i) imparting of information, (ii) interpersonal learning, (iii) catharsis, (iv) universality, (v) group cohesiveness, (vi) altruism, (vii) instillation of hope and (viii) existential factors. These domains illustrate how the therapeutic effects of group therapy worked to reduce patients' anxiety. CONCLUSION: The culturally sensitive body-mind-spirit group therapy reduced anxiety among outpatients with breast cancer. RELEVANCE TO CLINICAL PRACTICE: The involvement of mental health nurses in providing group therapy for cancer patients could enhance the quality of care in psycho-ontological nursing.

[Sequence analysis of the nucleocapsid gene and genome promoter region of peste des petits ruminants virus of Chinese origin].

The N gene and genome promoter nucleotide sequence of a Chinese Peste des petits rumiants virus (PPRV) ("China/Tib/Gej/07-30") was firstly determined. The length of N gene was 1689 nucleotides with a single open reading frame (ORF). The nucleotide and deduced amino acid sequence was compared with the homologous region of other PPRV isolates. The nucleotide sequence of the "China/Tib/Gej/07-30" was 91.7%-97.6% identical to other PPRV isolates, while a homology of 94.9%-98.5% could be observed at the amino acids level. The N gene encoded a protein of 525 amino acids. Several sequence motifs were identified on the basis of conservation in the PPRVs and the morbilliviruses. The genome length of promoter region was 107 nucleotides with 91.8%-98.2% identity to other PPRV isolates. Phylogenetic analysis showed that the "China/Tib/Gej/07-30" belonged to the Asian lineage.