Light-stabilized GIL1 suppresses PIN3 activity to inhibit hypocotyl gravitropism.

Light and gravity coordinately regulate the directional growth of plants. Arabidopsis Gravitropic in the Light 1 (GIL1) inhibits the negative gravitropism of hypocotyls in red and far-red light, but the underlying molecular mechanisms remain elusive. Our study found that GIL1 is a plasma membrane-localized protein. In endodermal cells of the upper part of hypocotyls, GIL1 controls the negative gravitropism of hypocotyls. GIL1 directly interacts with PIN3 and inhibits the auxin transport activity of PIN3. Mutation of PIN3 suppresses the abnormal gravitropic response of gil1 mutant. The GIL1 protein is unstable in darkness but it is stabilized by red and far-red light. Together, our data suggest that light-stabilized GIL1 inhibits the negative gravitropism of hypocotyls by suppressing the activity of the auxin transporter PIN3, thereby enhancing the emergence of young seedlings from the soil.

Amyloplast sedimentation repolarizes LAZYs to achieve gravity sensing in plants.

Gravity controls directional growth of plants, and the classical starch-statolith hypothesis proposed more than a century ago postulates that amyloplast sedimentation in specialized cells initiates gravity sensing, but the molecular mechanism remains uncharacterized. The LAZY proteins are known as key regulators of gravitropism, and lazy mutants show striking gravitropic defects. Here, we report that gravistimulation by reorientation triggers mitogen-activated protein kinase (MAPK) signaling-mediated phosphorylation of Arabidopsis LAZY proteins basally polarized in root columella cells. Phosphorylation of LAZY increases its interaction with several translocons at the outer envelope membrane of chloroplasts (TOC) proteins on the surface of amyloplasts, facilitating enrichment of LAZY proteins on amyloplasts. Amyloplast sedimentation subsequently guides LAZY to relocate to the new lower side of the plasma membrane in columella cells, where LAZY induces asymmetrical auxin distribution and root differential growth. Together, this study provides a molecular interpretation for the starch-statolith hypothesis: the organelle-movement-triggered molecular polarity formation.

Efficacy and safety of early enteral and intravenous fluid resuscitation in severe acute pancreatitis: a systematic review and meta-analysis.

OBJECTIVE: To compare the efficacy and safety of enteral fluid resuscitation (via nasointestinal tube or colorectal tube) and intravenous fluid resuscitation (via intravenous route) in the early treatment of severe acute pancreatitis. METHODS: In this study, 8 electronic databases (PubMed, Web of Science, Embase, Cochrane Library, Scopus, China HowNet database, Wanfang database, and VIP database) were searched to collect clinical studies from inception to June 12, 2022. After the quality evaluation and data extraction of the included studies, the RevMan 5.3 software was used for analysis. RESULTS: A total of seven studies including 580 patients were studied in this meta-analysis, in which 291 cases were treated with enteral fluid resuscitation and 289 cases were treated with intravenous fluid resuscitation. Compared with the intravenous route group, the enteral route resuscitation group reduced the incidence of new organ failure (OR = 0.23, 95% CI: 0.12-0.43, P < 0.00001), the incidence of persistent organ failure (OR = 0.38, 95% CI: 0.22-0.64, P = 0.0003), the incidence of mechanical ventilation (OR = 0.15, 95% CI: 0.03-0.69, P = 0.01), the incidence of ICU care (OR = 0.49, 95% CI: 0.27-0.88, P = 0.02), and the incidence of pancreatic infection (OR = 0.38, 95% CI: 0.17-0.83, P = 0.02). There were no statistically significant differences in mortality (OR = 0.77, 95% CI: 0.35-1.66, P = 0.50), surgical intervention rate (OR = 0.47, 95% CI: 0.19-1.18, P = 0.11), and incidence of localized ascites (OR = 0.65, 95% CI: 0.25-1.73, P = 0.39). CONCLUSION: Early enteral fluid resuscitation is safe and effective for in severe pancreatitis. But this conclusion needs to be verified by more additional multi-centre randomized controlled trials with large samples.

Thermally induced transformation of a Cu4I4-based cluster to a Cu2I2-based cluster under mild conditions.

A reaction of 6,6'-bis((benzylthio)methyl)-2,2'-bipyridine (L) with CuI at room temperature led to one Cu4I4-based cluster, which could be thermally transformed to a Cu2I2-based one under mild conditions due to the formation of a Cu-S bond. Along with the structural transformation, remarkable changes in the color and luminescence have been triggered.

Bottom-up and top-down factors of motion direction learning transfer.

Perceptual learning of motion discrimination has long been believed to be motion direction specific. However, recent studies using a double-training paradigm, in which the to-be-transferred condition was experienced through practicing an irrelevant task, found that perceptual learning in various visual tasks, including motion direction discrimination, can transfer completely to new conditions. This transfer occurred when the transfer stimulus was subconsciously presented, or when top-down attention was allocated to the transfer stimulus (which was absent). In the current study, observers were exposed subconsciously, or directed top-down attention, to the transfer motion direction, either simultaneously or successively with training. Data showed that motion direction learning transferred to the transfer direction, and suggest that motion direction learning specificity may result from under-activations of untrained visual neurons due to insufficient bottom-up stimulation and/or lack of top-down attention during training. These results shed new light on the neural mechanisms underlying motion perceptual learning and provide a constraint for models of motion perceptual learning.

Effects of genetically modified T2A-1 rice on the GI health of rats after 90-day supplement.

Bacillus thuringiensis insecticidal toxin (Bt) rice will be commercialized as a main food source. Traditional safety assessments on genetically modified products pay little attention on gastrointestinal (GI) health. More data about GI health of Bt rice must be provided to dispel public' doubts about the potential effects on human health. We constructed an improved safety assessment animal model using a basic subchronic toxicity experiment, measuring a range of parameters including microflora composition, intestinal permeability, epithelial structure, fecal enzymes, bacterial activity, and intestinal immunity. Significant differences were found between rice-fed groups and AIN93G-fed control groups in several parameters, whereas no differences were observed between genetically modified and non-genetically modified groups. No adverse effects were found on GI health resulting from genetically modified T2A-1 rice. In conclusion, this study may offer a systematic safety assessment model for GM material with respect to the effects on GI health.

The effect of genetically modified Lactobacillus plantarum 590 on the gut health of Sprague-Dawley rats.

Lp was a generally recognized as safe microorganism. Lactobacillus plantarum 590 was obtained by inserting nisI gene into Lp genome to help it tolerate higher concentration nisin. As the unintended effects of the genetically modified microorganism (GMM) are the most important barriers to the progress of GMM, we have performed a useful exploration to establish a new in vivo evaluation model for GMM from the point of view of intestinal health. In this study, Sprague-Dawley rats were orally administered with Lp 590 and Lp for 4 weeks. Fecal samples were collected to determine the number of beneficial bacteria Bifidobacterium and harmful bacteria Clostridium perfringens. Denaturing gradient gel electrophoresis was used to detect the bacterial profiles of every group. Fecal enzyme activities and short-chain fatty acids as main metabolites were also examined. Real time PCR (RT-PCR) and immunohistochemistry were used to analyze two proteins (ZO-1 and occludin) and secretory immunoglobulin A to detect intestinal permeability and mucosal immunity, gut permeability and gut mucosal immunity were analyzed to see whether GM Lp 590 can induce changes of the gut health when compared with non-GM Lp group, andeventually we concluded that there is no significant difference between GM Lp 590-fed group and non-GM Lp-fed group. The conclusion of gut health test was comparable withthat from traditional subchronic test. Evaluation of intestinal health will be a new approach of assessing the safety of GMM.

A novel universal primer-multiplex-PCR method with sequencing gel electrophoresis analysis.

In this study, a novel universal primer-multiplex-PCR (UP-M-PCR) method adding a universal primer (UP) in the multiplex PCR reaction system was described. A universal adapter was designed in the 5'-end of each specific primer pairs which matched with the specific DNA sequences for each template and also used as the universal primer (UP). PCR products were analyzed on sequencing gel electrophoresis (SGE) which had the advantage of exhibiting extraordinary resolution. This method overcame the disadvantages rooted deeply in conventional multiplex PCR such as complex manipulation, lower sensitivity, self-inhibition and amplification disparity resulting from different primers, and it got a high specificity and had a low detection limit of 0.1 ng for single kind of crops when screening the presence of genetically modified (GM) crops in mixture samples. The novel developed multiplex PCR assay with sequencing gel electrophoresis analysis will be useful in many fields, such as verifying the GM status of a sample irrespective of the crop and GM trait and so on.

Safety assessment of transgenic Bacillus thuringiensis rice T1c-19 in Sprague-Dawley rats from metabonomics and bacterial profile perspectives.

Bacillus thuringiensis rice is facing commercialization as the main food source in the near future. The unintended effects of genetically modified (GM) organisms are the most important barriers to their promotion. We aimed to establish a new in vivo evaluation model for genetically modified foods by using metabonomics and bacterial profile approaches. T1c-19 rice flour or its transgenic parent MH63 was used at 70% wt/wt to produce diets that were fed to rats for ∼ 90 days. Urine metabolite changes were detected using (1)H NMR. Denaturing gradient gel electrophoresis and real-time polymerase chain reaction (RT-PCR) were used to detect the bacterial profiles between the two groups. The metabonomics was analyzed for metabolite changes in rat urine, when compared with the non-GM rice group, where rats were fed a GM rice diet. Several metabolites correlated with rat age and sex but not with GM rice diet. Significant biological differences were not identified between the GM rice diet and the non-GM rice diet. The bacteria related to rat urine metabolites were also discussed. The results from metabonomics and bacterial profile analyses were comparable with the results attained using the traditional method. Because metabonomics and bacterial profiling offer noninvasive, dynamic approaches for monitoring food safety, they provide a novel process for assessing the safety of GM foods.

Metabonomics study of transgenic Bacillus thuringiensis rice (T2A-1) meal in a 90-day dietary toxicity study in rats.

Rice is one of the most important staple foods in the world. The Cry2A gene was inserted into the rice genome to help the plant combat insects. As the unintended effects of the genetically modified (GM) organisms are the most important barriers to the promotion of GM organisms, we have carried out a useful exploration to establish a new in vivo evaluation model for genetically modified foods by metabonomics methods. In this study, the rats were fed for 90 days with the GM and NON-GM rice diets. The changes in metabolites of the urine were detected using (1)H-NMR. The metabonomics were analyzed to see whether the GM rice can induce the metabolite changes in the rats' urine when compared with the NON-GM rice group. The multivariate analysis and ANOVA were used to determine the differences and the significance of differences respectively, and eventually we concluded that these differences did not have a biological significance. The conclusion of the metabonomics was comparable with that from the traditional method. As a non-invasive and dynamic monitoring method, metabonomics will be a new way of assessing the food safety of GM foods.

Effects of genetically modified T2A-1 rice on faecal microflora of rats during 90 day supplementation.

BACKGROUND: Many animal studies have been performed on products with the Bacillus thuringiensis insecticidal toxin-encoding gene (Bt products), but less have focused on its effects on intestinal microflora owing to difficulties in culturing. This 90 day study was designed to assess unintended effects of genetically modified T2A-1 rice (GMR) on selected intestinal bacteria (Lactobacillus group, Bifidobacterium genus, Escherichia coli subgroup, Enterococcus genus and Clostridium perfringens) of rats by the real-time polymerase chain reaction (PCR) method. RESULTS: During the whole experiment, no statistically significant differences in the numbers of specific bacteria and total bacteria were found between the GMR group and its parental group. At all stages of the experiment the two main probiotics (Lactobacillus group and Bifidobacterium genus) in faeces accounted for 11-23% of the total bacteria, whereas the conditional pathogens (E. coli subgroup, Enterococcus genus and C. perfringens) made up less than 1% of the total bacteria. B/E (log(10) copies of Bifidobacterium genome g(-1) faeces/log(10) copies of E. coli genome g(-1) faeces) ratios from 1.19 to 1.34 were obtained. Furthermore, significant correlations (P < 0.01) between the real-time PCR method and the plate count method were found, with r values ranging from 0.60 to 0.75. CONCLUSION: No adverse effects on the numbers of specific bacteria in rat faeces were observed as a result of GMR feeding. The real-time PCR method is recommended in further studies on the composition and dynamics of the intestinal bacteria community for better safety assessment of GM materials.

Assessment of tolerance to multistresses and in vitro cell adhesion in genetically modified Lactobacillus plantarum 590.

Lactobacillus plantarum (Lp) is a lactic acid bacterium that has many excellent traits that meet the needs of industrial production. Genetically modified (GM) Lp590 was obtained from Lp that was modified by the insertion of the gene nisI, which can confer resistance to nisin and play a role as a bio-preservative. Here, explorations were made to assess the safety of GM Lp590 and establish an in vitro evaluation model. The ability of Lp590 to tolerate both environmental stresses (such as temperatures ranging from 52 to 4 °C, or exposure to ethanol, oxygen, and osmotic stresses) and gastrointestinal transit was assessed. Lp590 showed a tolerance to 4 °C and ethanol (20%) within a period of 240 min that was similar to Lp. Notably, Lp590 can tolerate higher temperature (52 °C) and higher levels of H(2)O(2) (2%) and NaCl (4.0 M) than Lp. In contrast, Lp590 has the same gastrointestinal transit tolerance as Lp. In addition, Lp590 can adhere to Caco-2 cells, and it has no adverse effect on the cell membrane in vitro. These results indicate that GM Lp590 has many desirable biological characteristics and has good prospects for industrial applications. A useful and comprehensive exploration has been undertaken to establish a new in vitro evaluation model for genetically modified microorganisms (GMMs).

Safety assessment of Cry1C protein from genetically modified rice according to the national standards of PR China for a new food resource.

The Cry1C protein produced in Escherichia coli was used for in vitro evaluation and animal studies to support the safety assessment of GM food or feed products containing the Cry1C protein. The Cry1C protein does not have any sequence homology with known allergens or toxins. Although the Cry1C protein was heat stable it was rapidly degraded in vitro with simulated gastric or intestinal fluids. It did not cause adverse effects in mice as administered by gavage at a high level dosage of 5 g (Cry1C protein)/kg body weight. The mutagenicity of this protein was evaluated according to the national standards of People's Republic of China (PR China) for a new food resource. In mutagenic tests, the Cry1C protein caused<4 micronucleated cells per 1000 cells, <16 sperm abnormalities per 1000 cells and was not associated with any increased mutations in the Ames test. Taken together, these data indicate that the Cry1C protein is not a potential allergen or toxin.

Universal primer-multiplex PCR approach for simultaneous detection of Escherichia coli, Listeria monocytogenes, and Salmonella spp. in food samples.

Escherichia coli, Listeria monocytogenes, and Salmonella spp. are 3 kinds of the most important food-borne human pathogens. Traditional microbiological analysis is labor-intensive, time-consuming, and easily contaminated, thus producing false positive signals; it also involves much subjectivity judgments. Multiplex-PCR could be applied to detect multiple target organisms simultaneously to save time and labor, but there is always disproportionate amplification resulting from the disparity of different primers. To gain a rapid and sensitive method, a universal primer-multiplex PCR system (UP-M-PCR) was developed and applied for simultaneous detection of the 3 organisms. This method simplified traditional multiplex-PCR reaction system and overcame its amplification disparities among different primers; moreover, it got a high specificity and sensitivity (85, 155, and 104 copies/reaction for E. coli O157, L. monocytogenes, and Salmonella spp., respectively). Compared with the time-consuming and laborious microbiological analysis, UP-M-PCR had a lower risk of cross-contamination without inoculation and incubation. Test results for 36 food samples showed that UP-M-PCR method got a relative accuracy of 91.77% when compared with traditional microbiological analysis. It could serve as a rapid screening method for pathogen detection and could detect target genes even in dead pathogenic cells. In addition, it has the potential to be performed in an automation mode and might find broader application in simultaneous detection of other multiple pathogens.

Safety assessment of Cry1Ab/Ac fusion protein.

Cry1ab/ac gene was fused by both the cry1ab gene (GenBank Accession No. X54939) and the cry1ac gene (GenBank Accession No. Y09787), which was widely used in genetically modified (GM) rice, cotton, maize and so on. In order to support the safety assessment of GM food or feed products containing Cry1Ab/Ac protein, sufficient quantities of Cry1Ab/Ac protein were produced in Escherichia coli for in vitro evaluation and animal studies. The Cry1Ab/Ac protein does not possess the characteristics associated with food toxins or allergens, i.e., it has no sequence homology with any known allergens or toxins, and no N-glycosylation sites, can be rapidly degraded in gastric and intestinal fluids, and is devoid of adverse effects in mice by gavage at a high dose level of 5g (Cry1Ab/Ac protein)/kg body weight. In conclusion, there is a reasonable certainty of no harm resulting from the inclusion of the Cry1Ab/Ac protein in human food or animal feed.

Event-specific detection of stacked genetically modified maize Bt11 x GA21 by UP-M-PCR and real-time PCR.

More and more stacked GMOs have been developed for more improved functional properties and/or a stronger intended characteristic, such as antipest, improved product efficiency etc. Bt11 x GA21 is a new kind of stacked GM maize developed by Monsanto Company. Since there are no unique flanking sequences in stacked GMOs, up to now, no appropriate method has been reported to accurately detect them. In this passage, a novel universal primer multiplex PCR (UP-M-PCR) was developed and applied as a rapid screening method for the simultaneous detection of five target sequences (NOS, 35S, Bt11 event, GA21 event, and IVR) in maize Bt11 x GA21. This method overcame the disadvantages rooted deeply in conventional multiplex PCR such as complex manipulation, lower sensitivity, self-inhibition and amplification disparity resulting from different primers. What's more, it got a high specificity and had a detection limit of 0.1% (approximates to 38 haploid genome copies). Furthermore, real-time PCR combined with multivariate statistical analysis was used for accurate quantification of stacked GM maize Bt11 x GA21 in 100% GM maize mixture (Bt11 x GA21, Bt11 and GA21). Detection results showed that this method could accurately validate the content of Bt11, GA21 and Bt11 x GA21 in 100% GM mixture with a detection limit of 0.5% (approximates to 200 haploid genome copies) and a low relative standard deviation <5%. All the data proved that this method may be widely applied in event-specific detection of other stacked GMOs in GM-mixture.