The prospective associations among time management tendency, negative emotions, and problematic smartphone use in Chinese nursing students: enlightenment from COVID-19.

Purpose: The regularity of epidemic prevention and control measures in China has meant that nursing students have been exposed to more electronic devices, while problematic smartphone use has increased. The purpose of this study is to determine the prospective associations among time management tendency, negative emotions, and problematic smartphone use in nursing students during the COVID-19 pandemic. Methods: A longitudinal study was conducted between November 2021 and May 2022. A total of 989 nursing students participated. The convenience sampling method was adopted and the following tools were used: the Adolescence Time Management Disposition Scale, the Depression Anxiety Stress Scales - 21, and the Mobile Phone Addiction Index. Multiple parallel mediation models were used by Mplus. Results: Time management tendency had a significantly negative effect on problematic smartphone use (p < 0.05). Further tests using mediation models showed that stress as a negative emotion mediated the relationship between time management tendency and problematic smartphone use (p < 0.05) over time. Conclusion: Nursing educators need to strengthen the stress resistance and time management ability of nursing students.

A role of gut-brain axis on prophylactic actions of arketamine in male mice exposed to chronic restrain stress.

The gut-brain axis, which includes gut microbiota and microbiome-derived metabolites, might be implicated in depression. We reported the sustained prophylactic effects of a new antidepressant arketamine in chronic restrain stress (CRS) model of depression. In this study, we investigated the role of gut-brain axis on the prophylactic effects of arketamine in the CRS (7 days) model. Pretreatment with arketamine (10 mg/kg, 1 day prior to the CRS onset) significantly prevented CRS-induced body weight loss, increased immobility time of forced swimming test, decreased sucrose preference of sucrose preference test, and reduced expressions of synaptic proteins (GluA1 and PSD-95) in the prefrontal cortex (PFC) in the male mice. Gut microbiota analysis showed that pretreatment with arketamine might restore altered abundance of gut microbiota in CRS-exposed mice. An untargeted metabolomics analysis revealed four metabolites (e.g., L-leucine, N-acetyl-l-glutamine, 2-(2,4-dichlorophenyl)-3-[4-(dimethylamino)phenyl]acrylonitrile, L-threonine amide) that were altered between control and CRS group; however, there were found to be altered between the saline + CRS group and the arketamine + CRS group. Network analysis demonstrated correlations among synaptic proteins in the PFC and certain microbiota, and blood metabolites. These findings suggest that gut-brain axis, including its metabolites, might partially contribute to the persistent prophylactic effects of arketamine in the CRS model.

The influence of different sources of anticipated instrumental support on depressive symptoms in older adults.

Objective: This study investigated how anticipated instrumental support sources and intergenerational support influence depressive symptoms in older Chinese adults. Methods: We employed binary logistic regression on data from 7,117 adults aged ≥60 in the 2018 China Health and Retirement Longitudinal Study, controlling for gender, marital status, and self-rated health. Results: 38.89% of respondents exhibited depressive symptoms. Anticipated support from spouse and children, spouse only, children only, or other sources showed 52, 25, 46, and 40% lower odds of depression, respectively, compared with no anticipated support. Those providing financial support had 36% higher odds of depression than those without exchanges. However, those receiving financial support, receiving instrumental support, and receiving and providing financial and emotional support had 19, 14, 23, and 24% lower odds of depression. Conclusion: Different anticipated instrumental support sources and intergenerational support influenced depression odds in older adults, suggesting potential benefits in promoting such support systems.

Compact mode converter on SOI based on a polygonal subwavelength grating structure.

In this Letter, we design and experimentally demonstrate compact mode converters with a lightning-like and arrow-like polygonal subwavelength grating (SWG) structure on a silicon-on-insulator (SOI) platform, which can convert the TE0 mode to the TE1 and TE2 modes, respectively. The footprints of the proposed TE0-1 and TE0-2 mode converters are only 4.44 × 1.3 and 5.89 × 1.8 µm2, respectively. The experimental results show the mode converters have a low insertion loss (<1 dB) and a broad bandwidth (>50 nm). The measured cross talks of the TE0-1 and TE0-2 mode converters are -7.2 dB and -10.3 dB, respectively. In addition, the proposed mode converters with the SWG structure have the advantage in fabrication, since only a one-step full-etching process is required.

The influence of health literacy and knowledge about smoking hazards on the intention to quit smoking and its intensity: an empirical study based on the data of China's health literacy investigation.

OBJECTIVE: This study explored the relationship between smokers' health literacy, knowledge of smoking hazards, and their intention to quit. METHODS: Based on data from the 2019 Health Literacy and Tobacco Use Surveillance among residents of a city in Zhejiang Province, 1120 male smokers were screened. Differential tests were used to analyze whether smokers with varying levels of health literacy and knowledge about smoking hazards differed in their intention to quit smoking and the intensity of their intention. A multi-factor logistic regression model was constructed to explore the extent of these differences. RESULTS: Only 24.8% of smokers had higher health literacy. Among smokers, those with an intention to quit had a higher health literacy level compared to those without such intention (32.7% vs. 17.0%, p < 0.001). Health literacy levels did not differ significantly between groups with different intensity of intention to quit (34.2% vs. 31.9% vs. 30.1%, p = 0.435). About 48.7% of the smokers a higher level of knowledge about smoking hazards. It was more prevalent in the intent to quit group compared to the no intent to quit group (54.0% vs. 43.4%, p < 0.001), and the low intent to quit group had lower knowledge compared to the moderate and high intent to quit groups (49.1% vs. 56.6% vs. 63.4%, p = 0.011). After adjusting for other influences, smokers with lower health literacy were less likely to have intention to quit (OR = 0.659, p = 0.016). And the association between knowledge about smoking hazards and whether smokers have the intention to quit is no longer significant, but it significantly affects the intensity of the intention to quit among smokers who already have the intention (OR = 0.623, p = 0.005). CONCLUSION: General health literacy may play a role in facilitating smokers' progression from the stage of no intent to quit to one of intent to quit, but a more specific understanding of the harms of smoking may be needed to increase the strength of intent to quit.

Anion-π interactions suppress phase impurities in FAPbI3 solar cells.

Achieving both high efficiency and long-term stability is the key to the commercialization of perovskite solar cells (PSCs)1,2. However, the diversity of perovskite (ABX3) compositions and phases makes it challenging to fabricate high-quality films3-5. Perovskite formation relies on the reaction between AX and BX2, whereas most conventional methods for film-growth regulation are based solely on the interaction with the BX2 component. Herein, we demonstrate an alternative approach to modulate reaction kinetics by anion-π interaction between AX and hexafluorobenzene (HFB). Notably, these two approaches are independent but work together to establish 'dual-site regulation', which achieves a delicate control over the reaction between AX and BX2 without unwanted intermediates. The resultant formamidinium lead halides (FAPbI3) films exhibit fewer defects, redshifted absorption and high phase purity without detectable nanoscale δ phase. Consequently, we achieved PSCs with power conversion efficiency (PCE) up to 26.07% for a 0.08-cm2 device (25.8% certified) and 24.63% for a 1-cm2 device. The device also kept 94% of its initial PCE after maximum power point (MPP) tracking for 1,258 h under full-spectrum AM 1.5 G sunlight at 50 ± 5 °C. This method expands the range of chemical interactions that occur in perovskite precursors by exploring anion-π interactions and highlights the importance of the AX component as a new and effective working site to improved photovoltaic devices with high quality and phase purity.

Prophylactic effects of arketamine, but not hallucinogenic psychedelic DOI nor non-hallucinogenic psychedelic analog lisuride, in lipopolysaccharide-treated mice and mice exposed to chronic restrain stress.

Anesthetic ketamine and classical psychedelics that act as 5-hydroxytryptamine-2A receptor (5-HT2AR) agonists demonstrated rapid and sustained antidepressant actions in patients with treatment-resistant depression. The new antidepressant arketamine is reported to cause long-lasting prophylactic effects in lipopolysaccharide (LPS)-treated mice and mice exposed to chronic restrain stress (CRS). However, no study has compared the prophylactic effects of DOI (2,5-dimethoxy-4-iodoamphetamine: a hallucinogenic psychedelic drug with potent 5-HT2AR agonism), lisuride (non-hallucinogenic psychedelic analog with 5-HT2AR and 5-HT1AR agonism), and arketamine on depression-like behaviors in mice. Saline (10 ml/kg), DOI (2.0 or 4.0 mg/kg), lisuride (1.0 or 2.0 mg/kg), or arketamine (10 mg/kg) was administered intraperitoneally (i.p.) to male mice 6 days before administration of LPS (1.0 mg/kg). Pretreatment with aketamine, but not DOI and lisuride, significantly ameliorated body weight loss, splenomegaly, the increased immobility time of forced swimming test (FST), and the decreased expression of PSD-95 in the prefrontal cortex (PFC) of LPS-treated mice. In another test, male mice received the same treatment one day before CRS (7 days). Pretreatment with aketamine, but not DOI and lisuride, significantly ameliorated the increased FST immobility time, the reduced sucrose preference in the sucrose preference test, and the decreased expression of PSD-95 in the PFC of CRS-exposed mice. These findings suggest that, unlike to arketamine, both DOI and lisuride did not exhibit long-lasting prophylactic effects in mouse models of depression.

Anion Confinement for Homogeneous Mixed Halide Perovskite Film Growth by Electrospray.

Wide-bandgap perovskites are promising absorbers for state-of-the-art tandem solar cells to feasibly surpass Shockley-Queisser limit with low cost. However, the commonly used mixed halide perovskites suffer from poor stability; particularly, photoinduced phase segregation. Electrospray deposition is developed to bridge the gap of growth rate between iodide and bromide components during film growth by spatially confining the anion diffusion and eliminating the kinetic difference, which universally improves the initial homogeneity of perovskite films regardless of device architectures. It thus promotes the efficiency and stability of corresponding solar cells based on wide-bandgap (1.68 eV) absorbers. Remarkable power conversion efficiencies (PCEs) of 21.44% and 20.77% are achieved in 0.08 cm2 and 1.0 cm2 devices, respectively. In addition, these devices maintain 90% of their initial PCE after 1550 h of stabilized power output (SPO) tracking upon one sun irradiation (LED) at room temperature.

Discovery of spectacular quasar-driven superbubbles in red quasars.

Quasar-driven outflows on galactic scales are a routinely invoked ingredient for galaxy formation models. We report the discovery of ionized gas nebulae surrounding three luminous red quasars at z ~ 0.4 from Gemini integral field unit observations. All these nebulae feature unprecedented pairs of "superbubbles" extending ~20 kpc in diameter, and the line-of-sight velocity difference between the red- and blueshifted bubbles reaches up to ~1200 km/s. Their spectacular dual-bubble morphology (in analogy to the galactic "Fermi bubbles") and their kinematics provide unambiguous evidence for galaxy-wide quasar-driven outflows, in parallel with the quasi-spherical outflows similar in size from luminous type 1 and type 2 quasars at concordant redshift. These bubble pairs manifest themselves as a signpost of the short-lived superbubble "break-out" phase, when the quasar wind drives the bubbles to escape the confinement from the dense environment and plunge into the galactic halo with a high-velocity expansion.

Effects of esketamine on postoperative rebound pain in patients undergoing unilateral total knee arthroplasty: a single-center, randomized, double-blind, placebo-controlled trial protocol.

Introduction: Rebound pain, transient and acute postoperative pain after the disappearance of regional block anesthesia, has been a concern in recent years. Insufficient preemptive analgesia and hyperalgesia induced by regional block are the main mechanisms. At present, the evidence for the treatment of rebound pain is limited. The esketamine, as an antagonist of the N-methyl-D-aspartate receptor, has been proven to prevent hyperalgesia. Therefore, this trial aims to evaluate the impact of esketamine on postoperative rebound pain in patients undergoing total knee arthroplasty. Methods/design: This study is a single-center, prospective, double-blind, randomized, placebo-controlled trial. Participants who plan to undergo total knee arthroplasty will be randomly assigned to the esketamine group (N = 178) and placebo group (N = 178) in a ratio of 1:1. This trial aims to evaluate the impact of esketamine on postoperative rebound pain in patients undergoing total knee arthroplasty. The primary outcome of this trial is the incidence of rebound pain within 12 h after the operation in the esketamine group and the placebo group. The secondary outcome will be to compare (1) the incidence of rebound pain 24 h after the operation; (2) the time to enter the pain cycle for the first time within 24 h after the procedure; (3) the first time of rebound pain occurred within 24 h after surgery; (4) the modified rebound pain score; (5) NRS score under rest and exercise at different time points; (6) the cumulative opioid consumption at different time points; (7) patient's prognosis and knee joint function evaluation; (8) blood glucose and cortisol concentration; (9) patient's satisfaction score; (10) adverse reactions and adverse events. Discussion: The effect of ketamine on preventing postoperative rebound pain is contradictory and uncertain. The affinity of esketamine to the N-methyl-D-aspartate receptor is about four times higher than levo-ketamine, the analgesic effect is 3 times higher than levo-ketamine, and there are fewer adverse mental reactions. To our knowledge, there is no randomized controlled trial to verify the impact of esketamine on postoperative rebound pain in patients undergoing total knee arthroplasty. Therefore, this trial is expected to fill an important gap in relevant fields and provide novel evidence for individualized pain management. Clinical Trial Registration: http://www.chictr.org.cn, identifier ChiCTR2300069044.

Tungstate-mediated In-situ Passivation of Grain Boundary Grooves in Perovskite Solar Cells.

Possessed with advantageous optoelectronic properties, perovskites have boosted the rapid development of solution-processed solar cells. The performance of perovskite solar cells (PSCs) is significantly weakened by the carrier loss at grain boundary grooves (GBGs); however, it receives limited attention and there lacks effective approach to solve this issue. Herein, for the first time, we constructed the tungstate/perovskite heterointerface via a "two step" in situ reaction approach that provides effective defect passivation and ensures efficient carrier dynamics at the GBGs. The exposed perovskite at grain boundaries is converted to wide-band-gap PbWO4 via an in-situ reaction between Pb2+ and tungstate ions, which passivate defects due to the strong ionic bonding. Moreover, recombination loss is further suppressed via the heterointerface energetics modification based on an additional transformation from PbWO4 to CaWO4 . PSCs based on this groove modification strategy showed good universality in both normal and inverted structure, with an improved efficiency of 23.25 % in the n-i-p device and 23.33 % in the p-i-n device. Stable power output of the modified device could maintain 91.7 % after around 1100 h, and the device efficiency could retain 92.5 % after aging in air for around 2110 h, and 93.1 % after aging at 85 °C in N2 for 972 h.

Influence of health literacy on health outcomes of different social strata-- an empirical study based on the data of China's health literacy investigation.

BACKGROUND: Health literacy has always been considered as an important factor to promote people's health, but does it have a significant effect on health across all social strata and especially lower social strata? This study aims to analyze the influences of health literacy on health outcomes of different social strata, and then infer whether improving health literacy can reduce health disparities among different social strata. METHODS: Utilizing health literacy monitoring data from a city in Zhejiang Province in 2020, the samples are divided into three social strata according to the socioeconomic status score: low, middle and high social stratum, to compare whether there are significant differences in health outcomes between population with lower and higher health literacy among different social strata. In the strata with significant differences, control the confounding factors to further verify the influence of health literacy on health outcomes. RESULTS: In low and middle social strata, there are significant differences between population with lower and higher health literacy, when considering the two types of health outcomes (chronic diseases and self-rated health), but in high social stratum, this difference is not significant. After controlling the relevant variables, the influence of health literacy on the prevalence of chronic diseases is statistically significant only in low social stratum, and the health literacy is negatively correlated with the prevalence of chronic diseases(OR = 0.722, P = 0.022). In addition, there are statistical significances for positive impact of health literacy on self-rated health in both low and middle social strata (OR = 1.285, P = 0.047; OR = 1.401, P = 0.023). CONCLUSION: Compared with high social stratum, the influence of health literacy on health outcomes of low social stratum (chronic diseases) or both middle and low social strata (self-rated health) is more significant, and both are to improve the health outcomes. This finding suggests that improving residents' health literacy may be an effective way to alleviate the health disparities among different social strata.

The highly exothermic hydrogen abstraction reaction H2Te + OH → H2O + TeH: comparison with analogous reactions for H2Se and H2S.

The "gold standard" CCSD(T) method is adopted along with the correlation consistent basis sets up to aug-cc-pV5Z-PP to study the mechanism of the hydrogen abstraction reaction H2Te + OH. The predicted geometries and vibrational frequencies for reactants and products are in good agreement with the available experimental results. With the ZPVE corrections, the transition state in the favorable pathway of this reaction energetically lies 1.2 kcal mol-1 below the reactants, which is lower than the analogous relative energies for the H2Se + OH reaction (-0.7 kcal mol-1), the H2S + OH reaction (+0.8 kcal mol-1) and the H2O + OH reaction (+9.0 kcal mol-1). Accordingly, the exothermic reaction energies for these related reactions are predicted to be 47.8 (H2Te), 37.7 (H2Se), 27.1 (H2S), and 0.0 (H2O) kcal mol-1, respectively. Geometrically, the low-lying reactant complexes for H2Te + OH and H2Se + OH are two-center three-electron hemibonded structures, whereas those for H2S + OH and H2O + OH are hydrogen-bonded. With ZPVE and spin-orbit coupling corrections, the relative energies for the reactant complex, transition state, product complex, and the products for the H2Te + OH reaction are estimated to be -13.1, -1.0, -52.0, and -52.6 kcal mol-1, respectively. Finally, twenty-eight DFT functionals have been tested systematically to assess their ability in describing the potential energy surface of the H2Te + OH reaction. The best of these functionals for the corresponding energtics are -9.9, -1.4, -46.4, and -45.4 kcal mol-1 (MPWB1K), or -13.1, -2.4, -57.1, and -54.6 kcal mol-1 (M06-2X), respectively.

Study of resonant mode coupling in the transverse-mode-conversion based resonator with an anti-symmetric nanobeam Bragg reflector.

The traveling-wave like Fabry-Perot (F-P) resonators based on transverse-mode-conversion have been extensively studied as on-chip filters. However, the incomplete transverse mode conversion will lead to the coupling between two degenerated resonant modes, which brings additional loss and may further induce the resonance splitting. In this paper, we take the transverse-mode-conversion based resonator with anti-symmetric nanobeam Bragg reflector as an example and study the resonant mode coupling in both the direct-coupled and side-coupled resonators. The coupled mode equations are used to model the incomplete transverse mode conversion of Bragg reflector. The resonant mode coupling can be effectively suppressed by carefully designing the phase shifter length and adding the tapered holes. The insertion loss of less than -1 dB can be achieved in the simulation using the two methods. This work is believed to benefit the design of mode-conversion based resonators with low insertion loss and non-splitting line shape.

Eukaryotic plankton community assembly and influencing factors between continental shelf and slope sites in the northern South China Sea.

Eukaryotic plankton are pivotal members of marine ecosystems playing crucial roles in marine food webs and biogeochemical cycles. However, understanding the patterns and drivers of their community assembly remains a grand challenge. A study was conducted in the northern South China Sea (SCS) to address this issue. Here, 49 samples were collected and size-fractionated from discrete depths at continental shelf and continental slope in the northern SCS over a diel cycle. From high throughput sequencing of the 18S rDNA gene V4 region, 2463 operational taxonomic units (OTUs) were retrieved. Alveolata and Opisthokonta overwhelmingly dominated the assemblages in the abundance (44.76%, 31.08%) and species richness (59%, 12%). Biodiversity was higher in the slope than the shelf and increased with depth. Temperature and salinity appeared to be the most important deterministic drivers of taxon composition. Community structure was influenced by multiple factors in the importance order of: environmental factors (temperature + salinity) > spatial factor > water depth > sampling time. Furthermore, the neutral model explained more variations in the smaller-sized (0.22-3 µm) community (24%) than larger-sized (3-200 µm) community (16%) but generally explained less variations than did deterministic processes. Additionally, our data indicated that the larger plankton might be more environmentally filtered and less plastic whereas the smaller plankton had stronger dispersal ability. This study sheds light on the differential contributions of the deterministic process and stochastic process and complexities of assembly mechanisms in shaping the community assembly of micro-nano and pico-eukaryotic biospheres in a subtropical ocean.

Partial Convolution for Padding, Inpainting, and Image Synthesis.

Partial convolution weights convolutions with binary masks and renormalizes on valid pixels. It was originally proposed for image inpainting task because a corrupted image processed by a standard convolutional often leads to artifacts. Therefore, binary masks are constructed that define the valid and corrupted pixels, so that partial convolution results are only calculated based on valid pixels. It has been also used for conditional image synthesis task, so that when a scene is generated, convolution results of an instance depend only on the feature values that belong to the same instance. One of the unexplored applications for partial convolution is padding which is a critical component of modern convolutional networks. Common padding schemes make strong assumptions about how the padded data should be extrapolated. We show that these padding schemes impair model accuracy, whereas partial convolution based padding provides consistent improvements across a range of tasks. In this article, we review partial convolution applications under one framework. We conduct a comprehensive study of the partial convolution based padding on a variety of computer vision tasks, including image classification, 3D-convolution-based action recognition, and semantic segmentation. Our results suggest that partial convolution-based padding shows promising improvements over strong baselines.

The Investigation of the Influence of a Cu2O Buffer Layer on Hole Transport Layers in MAPbI3-Based Perovskite Solar Cells.

The passivation engineering of the hole transport layer in perovskite solar cells (PSCs) has significantly decreased carrier accumulation and open circuit voltage (Voc) loss, as well as energy band mismatching, thus achieving the goal of high-power conversion efficiency. However, most devices incorporating organic/inorganic buffer layers suffer from poor stability and low efficiency. In this article, we have proposed an inorganic buffer layer of Cu2O, which has achieved high efficiency on lower work function metals and various frequently used hole transport layers (HTLs). Once the Cu2O buffer layer was applied to modify the Cu/PTAA interface, the device exhibited a high Voc of 1.20 V, a high FF of 75.92%, and an enhanced PCE of 22.49% versus a Voc of 1.12 V, FF of 69.16%, and PCE of 18.99% from the (PTAA/Cu) n-i-p structure. Our simulation showed that the application of a Cu2O buffer layer improved the interfacial contact and energy alignment, promoting the carrier transportation and reducing the charge accumulation. Furthermore, we optimized the combinations of the thicknesses of the Cu2O, the absorber layer, and PTAA to obtain the best performance for Cu-based perovskite solar cells. Eventually, we explored the effect of the defect density between the HTL/absorber interface and the absorber/ETL interface on the device and recommended the appropriate reference defect density for experimental research. This work provides guidance for improving the experimental efficiency and reducing the cost of perovskite solar cells.

Effect of intergenerational exchange patterns and intergenerational relationship quality on depressive symptoms in the elderly: An empirical study on CHARLS data.

Background: As the population ages with fewer children, depression symptoms are increasing among the elderly who lack companionship. Intergenerational support is closely related to depression in the elderly; hence how the behavioral patterns and emotional quality of intergenerational support affect depressive symptoms in the elderly should be further explored. Objective: To study the effects of intergenerational exchange patterns and intergenerational relationship quality on depressive symptoms in the elderly. Methods: A total of 8,015 people over 60 years old in CHARLS in 2018 were selected as the object of this study. First, the correlation between demographics, economic conditions, health status, intergenerational support patterns, intergenerational relationship quality, and depressive symptoms in the elderly were analyzed. Three regression analysis models were established to analyze the relationship between control variables, intergenerational support patterns, intergenerational relationship quality, and depressive symptoms in the elderly. Results: Among the intergenerational economic, care, and emotional exchange modes, the risk of depressive symptoms in the elderly in the mutual support group was 31.8, 38.4, and 25.5% lower than that in the non-communication group. Compared with the elderly with very poor intergenerational relationship quality, the elderly with good, very good, and excellent intergenerational relationship quality had 74.5, 84.0, and 85.6% lower risk of depressive symptoms. Discussion: Different cultural backgrounds also affect intergenerational exchange patterns and depression in the elderly. During the study of depressive symptoms, two aspects relating to intergenerational support should be considered behaviorally and emotionally: the intergenerational exchange model and the intergenerational relationship quality. As depression in the elderly is affected by multiple factors, the participation and joint efforts of the whole society are required to reduce depressive symptoms in the elderly and realize active aging. Conclusion: The intergenerational exchange pattern of mutual support and the higher quality of the intergenerational relationship can significantly reduce the depressive symptoms of the elderly.

Sequencing introduced false positive rare taxa lead to biased microbial community diversity, assembly, and interaction interpretation in amplicon studies.

BACKGROUND: Increasing studies have demonstrated potential disproportionate functional and ecological contributions of rare taxa in a microbial community. However, the study of the microbial rare biosphere is hampered by their inherent scarcity and the deficiency of currently available techniques. Sample-wise cross contaminations might be introduced by sample index misassignment in the most widely used metabarcoding amplicon sequencing approach. Although downstream bioinformatic quality control and clustering or denoising algorithms could remove sequencing errors and non-biological artifact reads, no algorithm could eliminate high quality reads from sample-wise cross contaminations introduced by index misassignment, making it difficult to distinguish between bona fide rare taxa and potential false positives in metabarcoding studies. RESULTS: We thoroughly evaluated the rate of index misassignment of the widely used NovaSeq 6000 and DNBSEQ-G400 sequencing platforms using both commercial and customized mock communities, and observed significant lower (0.08% vs. 5.68%) fraction of potential false positive reads for DNBSEQ-G400 as compared to NovaSeq 6000. Significant batch effects could be caused by stochastically introduced false positive or false negative rare taxa. These false detections could also lead to inflated alpha diversity of relatively simple microbial communities and underestimated that of complex ones. Further test using a set of cow rumen samples reported differential rare taxa by different sequencing platforms. Correlation analysis of the rare taxa detected by each sequencing platform demonstrated that the rare taxa identified by DNBSEQ-G400 platform had a much higher possibility to be correlated with the physiochemical properties of rumen fluid as compared to NovaSeq 6000 platform. Community assembly mechanism and microbial network correlation analysis indicated that false positive or negative rare taxa detection could lead to biased community assembly mechanism and identification of fake keystone species of the community. CONCLUSIONS: We highly suggest proper positive/negative/blank controls, technical replicate settings, and proper sequencing platform selection in future amplicon studies, especially when the microbial rare biosphere would be focused.

Simultaneous quantification of marine neutral neoagaro-oligosaccharides and agar-oligosaccharides by the UHPLC-MS/MS method: application to the intestinal transport study by using the Caco-2 cell monolayer.

A sensitive and robust ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was established for the first time to simultaneously quantify marine neutral neoagaro-oligosaccharides (NAOS) and agar-oligosaccharides (AOS) with different degrees of polymerization (DP) in Hanks' balanced salt solution (HBSS). The separation was achieved on a BEH amide column using a mobile phase of acetonitrile-10 mmol L-1 ammonium acetate (58 : 42, v/v) with an isocratic elution program. The total analysis time was 3.5 min. The mass spectra were acquired in the multiple reaction monitoring (MRM) pattern by using a heated-electrospray ionization (H-ESI) source operating in the positive ionization mode. The linear range was 40-20 000 nmol L-1. The accuracy and precision ranged from 91.5 to 110.0% and 0.9 to 10.4%, respectively. The extraction recovery was consistent and reproducible. The stability was within 90.3-110.8%. The matrix effect, carryover, and dilution integrity were all satisfactory. Moreover, the validated method was successfully applied to the intestinal transport study by using the Caco-2 cell monolayer in vitro. The results revealed that neoagarobiose, neoagarotetraose, neoagarohexaose, agarotriose, agaropentose, and agaroheptose were transported by a paracellular pathway.