Rationally designed ß-cyclodextrin-crosslinked polyacrylamide hydrogels for cell spheroid formation and 3D tumor model construction.

In vitro tumor models are essential for understanding tumor behavior and evaluating tumor biological properties. Hydrogels that can mimic the tumor extracellular matrix have become popular for creating 3D in vitro tumor models. However, designing biocompatible hydrogels with appropriate chemical and physical properties for constructing tumor models is still a challenge. In this study, we synthesized a series of ß-cyclodextrin (ß-CD)-crosslinked polyacrylamide hydrogels with different ß-CD densities and mechanical properties and evaluated their potential for use in 3D in vitro tumor model construction, including cell capture and spheroid formation. By utilizing a combination of ß-CD-methacrylate (CD-MA) and a small amount of N,N'-methylene bisacrylamide (BIS) as hydrogel crosslinkers and optimizing the CD-MA/BIS ratio, the hydrogels performed excellently for tumor cell 3D culture and spheroid formation. Notably, when we co-cultured L929 fibroblasts with HeLa tumor cells on the hydrogel surface, co-cultured spheroids were formed, showing that the hydrogel can mimic the complexity of the tumor extracellular matrix. This comprehensive investigation of the relationship between hydrogel mechanical properties and biocompatibility provides important insights for hydrogel-based in vitro tumor modeling and advances our understanding of the mechanisms underlying tumor growth and progression.

Genome-wide identification and expression analysis of the ALDH gene family and functional analysis of PaALDH17 in Prunus avium.

ALDH (Aldehyde dehydrogenase), as an enzyme that encodes the dehydroxidization of aldehydes into corresponding carboxylic acids, played an important role inregulating gene expression in response to many kinds of biotic and abiotic stress, including saline-alkali stress. Saline-alkali stress was a common stress that seriously affected plant growth and productivity. Saline-alkali soil contained the characteristics of high salinity and high pH value, which could cause comprehensive damage such as osmotic stress, ion toxicity, high pH, and HCO3-/CO32- stress. In our study, 18 PaALDH genes were identified in sweet cherry genome, and their gene structures, phylogenetic analysis, chromosome localization, and promoter cis-acting elements were analyzed. Quantitative real-time PCR confirmed that PaALDH17 exhibited the highest expression compared to other members under saline-alkali stress. Subsequently, it was isolated from Prunus avium, and transgenic A. thaliana was successfully obtained. Compared with wild type, transgenic PaALDH17 plants grew better under saline-alkali stress and showed higher chlorophyll content, Superoxide dismutase (SOD), Peroxidase (POD) and Catalase (CAT) enzyme activities, which indicated that they had strong resistance to stress. These results indicated that PaALDH17 improved the resistance of sweet cherries to saline-alkali stress, which in turn improved quality and yields. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-024-01444-7.

Microecology of aerobic denitrification system construction driven by cyclic stress of sulfamethoxazole.

The construction of aerobic denitrification (AD) systems in an antibiotic-stressed environment is a serious challenge. This study investigated strategy of cyclic stress with concentration gradient (5-30 mg/L) of sulfamethoxazole (SMX) in a sequencing batch reactor (SBR), to achieve operation of AD. Total nitrogen removal efficiency of system increased from about 10 % to 95 %. Original response of abundant-rare genera to antibiotics was changed by SMX stress, particularly conditionally rare or abundant taxa (CRAT). AD process depends on synergistic effect of heterotrophic nitrifying aerobic denitrification bacteria (Paracoccus, Thauera, Hypomicrobium, etc). AmoABC, napA, and nirK were functionally co-expressed with multiple antibiotic resistance genes (ARGs) (acrR, ereAB, and mdtO), facilitating AD process. ARGs and TCA cycling synergistically enhance the antioxidant and electron transport capacities of AD process. Antibiotic efflux pump mechanism played an important role in operation of AD. The study provides strong support for regulating activated sludge to achieve in situ AD function.

Effects of Salt Stress on Salt-Repellent and Salt-Secreting Characteristics of Two Apple Rootstocks.

The effects of NaCl-induced salinity on biomass allocation, anatomical characteristics of leaves, ion accumulation, salt repellency, and salt secretion ability were investigated in two apple rootstock cultivars (Malus halliana '9-1-6' and Malus baccata), which revealed the physiological adaptive mechanisms of M. halliana '9-1-6' in response to salt stress factors. This experiment was conducted in a greenhouse using a nutrient solution pot. Salt stress was simulated by treating the plants with a 100 mM NaCl solution, while 1/2 Hoagland nutrient solution was used as a control (CK) instead of the NaCl solution. The results showed that the two rootstocks responded to salt environments by increasing the proportion of root biomass allocation. According to the stress susceptibility index, '9-1-6' exhibits a lower salt sensitivity index and a higher salt tolerance index. The thickness of the leaf, upper and lower epidermis, palisade tissue, and mesophyll tissue compactness (CTR) of the two rootstocks were significantly decreased, while the thickness of sponge tissue and mesophyll tissue looseness (SR) were significantly increased, and the range of '9-1-6' was smaller than that of M. baccata. With an extension of stress time, the accumulation of Na+ increased significantly, and the accumulation of K+ decreased gradually. The stem and leaves of '9-1-6' showed a lower accumulation of Na+ and a higher accumulation of K+, and the roots displayed a higher ability to reject Na+, as well as young and old leaves showed a stronger ability to secrete Na+. In conclusion, within a certain salt concentration range, the '9-1-6' root part can maintain lower salt sensitivity and a higher root-to-shoot ratio by increasing the proportion of root biomass allocation; the aerial part responds to salt stress through thicker leaves and a complete double-layer fence structure; the roots and stem bases can effectively reduce the transportation of Na+ to the aerial parts, as well as effectively secrete Na+ from the aerial parts through young and old leaves, thereby maintaining a higher K+/Na+ ratio in the aerial parts, showing a strong salt tolerance.

SPHYNCS: Feasibility of long-term monitoring with Fitbit smartwatches in central disorders of hypersomnolence and extraction of digital biomarkers in narcolepsy.

The Swiss Primary Hypersomnolence and Narcolepsy Cohort Study (SPHYNCS) is a multicenter research initiative to identify new biomarkers in central disorders of hypersomnolence (CDH). Whereas narcolepsy type 1 (NT1) is well characterized, other CDH disorders lack precise biomarkers. In SPHYNCS, we utilized Fitbit smartwatches to monitor physical activity, heart rate, and sleep parameters over one year. We examined the feasibility of long-term ambulatory monitoring using the wearable device. We then explored digital biomarkers differentiating patients with NT1 from healthy controls (HC). A total of 115 participants received a Fitbit smartwatch. Using a compliance metric to evaluate the usability of the wearable device, we found an overall compliance rate of 80% over one year. We calculated daily physical activity, heart rate, and sleep parameters from two weeks of greatest compliance to compare NT1 (n=20) and HC (n=9) subjects. Compared to controls, NT1 patients demonstrated findings consistent with increased sleep fragmentation, including significantly greater wake-after-sleep onset (p=0.007) and awakening index (p=0.025), as well as standard deviation of time in bed (p=0.044). Moreover, NT1 patients exhibited a significantly shorter REM latency (p=0.019), and sleep latency (p=0.001), as well as a lower peak heart rate (p=0.008), heart rate standard deviation (p=0.039) and high-intensity activity (p=0.009) compared to HC. This ongoing study demonstrates the feasibility of long-term monitoring with wearable technology in patients with CDH and potentially identifies a digital biomarker profile for NT1. While further validation is needed in larger datasets, these data suggest that long-term wearable technology may play a future role in diagnosing and managing narcolepsy.

Potential immunological triggers for narcolepsy and idiopathic hypersomnia: Real-world insights on infections and influenza vaccinations.

OBJECTIVE: It is hypothesized that narcolepsy type 1 (NT1) develops in genetically susceptible people who encounter environmental triggers leading to immune-mediated hypocretin-1 deficiency. The pathophysiologies of narcolepsy type 2 (NT2) and idiopathic hypersomnia (IH) remain unknown. The main aim of this study was to collect all reported immunological events before onset of a central disorder of hypersomnolence. METHODS: Medical records of 290 people with NT1, and 115 with NT2 or IH were retrospectively reviewed to extract infection and influenza vaccination history. Prevalence, distribution of immunological events, and time until hypersomnolence onset were compared between NT1 and the combined group of NT2 and IH. RESULTS: Immunological events were frequently reported before hypersomnolence disorder onset across groups. Flu and H1N1 influenza vaccination were more common in NT1, and Epstein-Barr virus and other respiratory and non-respiratory infections in NT2 and IH. Distributions of events were comparable between NT2 and IH. Rapid symptom onset within one month of infection was frequent across groups, especially after flu infection in NT1. Hypersomnolence disorder progression after an immunological event was reported in ten individuals. CONCLUSIONS: Our findings suggest a variety of immunological triggers potentially related to NT1, including H1N1 influenza infection or vaccination, infection with other flu types, and other respiratory and non-respiratory infections. Frequent reports of immunological events (other than those reported in NT1) immediately prior to the development of NT2 and IH support the specificity of triggers for NT1, and open important new research avenues into possible underlying immunological mechanisms in NT2 and IH.

Efficacy and safety of a panthenol-enriched mask for individuals with distinct impaired skin barrier subtypes.

BACKGROUND: The protection for different skin types with impaired skin barrier in the market is insufficient. AIM: To evaluate the efficacy and safety of a panthenol-enriched mask (La Roche-Posay Mask Pro) in addressing various skin barrier impairment subgroups, including dry sensitive, oily sensitive, and oily acne skin. METHODS: A total of 177 participants were enrolled in the study and divided into three subgroups based on their skin type. Participants used the mask following the specified protocol, with measurements taken for skin hydration, transepidermal water loss (TEWL), sebum content, and skin redness-factors that are directly influenced by skin barrier function. Assessments were conducted at baseline and after 1 day (tested 15 min post-application), 7 days, and 14 days of application using Sebumeter, Tewameter, Corneometer, Mexameter, and VISIA. RESULTS: Results showed significant improvements in skin parameters across all subgroups. In the dry sensitive skin subgroup, the mask increased skin hydration, sebum content, and reduced redness. For the oily sensitive skin subgroup, the mask regulated sebum production and improved skin hydration. In the oily acne skin subgroup, the mask reduced sebum content, redness, TEWL, and post-inflammatory erythema and hyperpigmentation. Tolerance was excellent for all skin types, with no adverse reactions observed. CONCLUSIONS: This study highlights the efficacy and safety of the panthenol-enriched LRP Mask Pro for individuals with distinct skin barrier impairment subgroups. The mask's versatile formulation and proven efficacy make it a valuable skincare product for addressing various skin concerns and achieving healthier, more balanced skin.

Efficacy of a moisturizing cream and facial mask for alleviating skin problems associated with medical mask use: A half-face, randomized controlled study.

BACKGROUND: Prolonged use of medical masks has increased skin-related issues. AIM: To evaluate the efficacy of a facial cream and facial mask in mitigating medical mask related skin symptoms. METHODS: Healthy women were randomly assigned to apply a facial cream (n = 32) or a facial mask plus a facial cream (n = 32) on half-faces after wearing medical masks for 4 h (Tb). Transepidermal water loss (TEWL), dryness score, and redness area were assessed at Tb and 10 min after using the cream (T1) in the facial cream group, and at Tb, 1 h after using the facial mask (T2), and 10 min after using the cream (T3) in the combined use group. RESULTS: In the facial cream group, the treated half-face showed significantly better improvements from Tb to T1 in TEWL (-2.95 ± 0.38 vs. -0.68 ± 0.35 g/h·cm2, p < 0.001) and skin dryness score (-1.00 ± 0.12 vs. 0.00 ± 0.00, p < 0.001). In the combined use group, the treated half-face showed significantly better improvements from Tb to T2 and T3 in TEWL (T2, -3.46 ± 0.33 vs. -0.09 ± 0.13 g/h·cm2; T3, -4.67 ± 0.31 vs. -0.28 ± 0.22 g/h·cm2) and skin dryness score (T2, -0.63 ± 0.13 vs. 0.03 ± 0.03; T3, -0.94 ± 0.17 vs. 0.19 ± 0.07) (all p < 0.001) then the untreated half-face. The combined use group had significantly lower TEWL at T3 than T2 (p < 0.05). The reduction in redness area was similar between the treated and untreated half-faces in both groups. CONCLUSIONS: The test facial cream and mask significantly improved skin barrier function and alleviated dryness symptoms associated with medical mask use, with the combined use offering superior benefits.

Evaluation of the efficacy of a sunscreen containing ultra-long UVA1 and other UVR broad-spectrum filters on skin barrier protection and melanin content reduction in Chinese adults: A single-center study.

Background and Aims: The protection for ultra-long UVA1 is lacked in the market, posing potential damage from ultra-long UVA1 irradiation. The study aims to evaluate the efficacy of a sunscreen containing multiple components, especially Mexoryl® 400 for improving skin barrier function and reducing melanin content. Methods: This single-center study included adults with sensitive and normal skin in China in November 2022. Participants received the test sunscreen for 4 weeks. Melanin and hemoglobin content, sebum secretion skin hydration, and trans-epidermal water loss were evaluated at T0d, T7d, and T28d. The self-assessment was done at T15min, T7d, and T28d. Results: Sixty participants were included, including 30 self-claimed sensitive skin in the sunscreen group. The sunscreen demonstrated significant improvements in skin parameters. Skin redness reduced by 9.84% at T28d, sebum content in the forehead area decreased by 22.70% at T28d, and skin stratum corneum hydration increased by 38.44% at T28d, p < 0.001 respectively. Most notably, skin melanin content significantly reduced by 13.49% after 4 weeks' usage (p < 0.001). No adverse reactions were reported in either group. Conclusions: The study sunscreen improved the skin condition by decreasing the melanin content, regulating skin barrier function, and achieving a balance of skin hydration and sebum secretion.

Efficacy and safety of a cream containing panthenol, prebiotics, and probiotic lysate for improving sensitive skin symptoms.

BACKGROUND: Sensitive skin is a common condition affecting a significant proportion of the population, and there is a growing demand for effective and safe management. AIM: To evaluate the efficacy and safety of a cream containing panthenol, prebiotics, and probiotic lysate as an optimal care for facial sensitive skin. METHODS: A total of 110 participants (64 in group A and 46 in group B) with facial sensitive skin applied the cream twice daily for 28 days. Group A evaluated their sensitive skin, product efficacy, and product use experience at D0 (15 min), D1, D14, and D28. In group B, skin barrier function-related indicators were measured at baseline and on D1, D7, D14, and D28. Dermatologists evaluated tolerance for all participants. RESULTS: After 28 days of use, in group A, 100% of participants reported mildness and comfort with product use. Participants demonstrated significant improvements in skin barrier function-related indicators, including increased stratum corneum moisture content, reduced erythema index, elevated sebum content, decreased trans-epidermal water loss, and diminished skin redness parameter a* value (all p < 0.05). Dermatologist evaluations revealed excellent tolerance among all participants. CONCLUSION: The panthenol-enriched cream with prebiotics and probiotic lysate exhibited substantial clinical efficacy in ameliorating facial sensitive skin conditions, coupled with a high safety profile.

Cumulative brain desaturation: Time to consider brain derived parameters to measure daytime sleepiness in obstructive sleep apnea.

OBJECTIVE: /Background: The change in cerebral hemodynamics induced by sleep apneas and hypopneas may contribute to the daytime sleepiness in patients with obstructive sleep apnea (OSA). However, previous studies failed to discovery their relationship. We propose and test a new parameter, the cumulative brain oxygen desaturation, which may contribute to OSA patient's daytime sleepiness. PATIENTS/METHODS: 22 patients with severe OSA (apnea-hypopnea index (AHI) at diagnosis [mean ± standard deviation, std.]: 52.1 ± 21.6/h, median: 45.1/h, interquartile range: 34.4-60.2/h) were monitored by polysomnography during routine continuous positive airway pressure titration. The reductions of brain tissue oxygen saturation (StO2) in all respiratory events at baseline sleep were measured by frequency-domain near-infrared spectroscopy (NIRS). The cumulative brain desaturation was calculated as AHI times the mean StO2 desaturation (i.e., AHI×ΔStO2‾). Similarly, cumulative peripheral desaturation was also calculated, i.e., AHI×ΔSpO2‾ where ΔSpO2‾ was the mean reduction of peripheral arterial oxygen saturation (SpO2). The correlations between Epworth sleepiness scale (ESS) and AHI, ΔStO2‾, AHI×ΔStO2‾, and AHI×ΔSpO2‾ were tested, respectively. Linear regression was applied to predict ESS using AHI×ΔStO2‾ and AHI×ΔSpO2‾, with age and BMI as covariates. RESULTS: ESS significantly correlates to the cumulative brain desaturation (Pearson's correlation coefficient: 0.68, p = 0.00056), not the other parameters. Regression analysis only finds significant association between ESS and the cumulative cerebral desaturation (p = 0.00195) but not the cumulative peripheral desaturation (p = 0.71). CONCLUSIONS: The cumulative brain oxygen desaturation, which comprehensively combines total sleep time, the frequency of apnea and hypopnea events, and the severity of cerebral oxygen desaturation, is a new indicator for daytime sleepiness in severe OSA.

Comprehensive analysis revealed that titanium dioxide nanoparticles could strengthen the resistance of apple rootstock B9 to saline-alkali stress.

Apple growth and development can be adversely affected by saline-alkali stress, which has become a significant factor restricting the high yield of the apple industry. In recent years, nanomaterials have become a potential source for plant growth and development. Titanium dioxide nanoparticles (TiO2 NPs) play an important role in multiple plant development processes, including mitigating environmental stress. In this study, one-year-old apple rootstock B9 stem cuttings were used as research objects. Different concentrations of TiO2 NPs were applied to the roots before saline-alkali treatment. Principal component analysis showed that 1gkg-1 TiO2 NPs treatment had the best effect in alleviating the stress for B9. It significantly reduced the damage to B9 under salt-alkali stress, increased the content of photosynthetic pigment, enhanced the performance of Photosystem II, and promoted photosynthesis. At the same time, the content of K+ was increased, and the ion toxicity was alleviated. In addition, TiO2 NPs have also been shown to reduce B9 cell damage and lipid peroxidation, increase antioxidant enzyme activity, and regulate the accumulation of solutes. Overall, this study provides a theoretical basis for TiO2 NPs to mitigate the adverse effects of plants under saline-alkali stress and provides useful insights for managing other plants affected by global salinity and alkalinity.

Transcription factors dealing with Iron-deficiency stress in plants: focus on the bHLH transcription factor family.

Iron (Fe), as an important micronutrient element necessary for plant growth and development, not only participates in multiple physiological and biochemical reactions in cells but also exerts a crucial role in respiration and photosynthetic electron transport. Since Fe is mainly present in the soil in the form of iron hydroxide, Fe deficiency exists universally in plants and has become an important factor triggering crop yield reduction and quality decline. It has been shown that transcription factors (TFs), as an important part of plant signaling pathways, not only coordinate the internal signals of different interaction partners during plant development, but also participate in plant responses to biological and abiotic stresses, such as Fe deficiency stress. Here, the role of bHLH transcription factors in the regulation of Fe homeostasis (mainly Fe uptake) is discussed with emphasis on the functions of MYB, WRKY and other TFs in the maintenance of Fe homeostasis. This review provides a theoretical basis for further studies on the regulation of TFs in Fe deficiency stress response.

The Comparison of Early Hemodynamic Response to Single-Pulse Transcranial Magnetic Stimulation following Inhibitory or Excitatory Theta Burst Stimulation on Motor Cortex.

We present a new study design aiming to enhance the understanding of the mechanism by which continuous theta burst stimulation (cTBS) or intermittent theta burst stimulation (iTBS) paradigms elicit cortical modulation. Using near-infrared spectroscopy (NIRS), we compared the cortical hemodynamics of the previously inhibited (after cTBS) or excited (after iTBS) left primary motor cortex (M1) as elicited by single-pulse TMS (spTMS) in a cross-over design. Mean relative changes in hemodynamics within 6 s of the stimulus were compared using a two-sample t-test (p < 0.05) and linear mixed model between real and sham stimuli and between stimuli after cTBS and iTBS. Only spTMS after cTBS resulted in a significant increase (p = 0.04) in blood volume (BV) compared to baseline. There were no significant changes in other hemodynamic parameters (oxygenated/deoxygenated hemoglobin). spTMS after cTBS induced a larger increase in BV than spTMS after iTBS (p = 0.021) and sham stimulus after cTBS (p = 0.009). BV showed no significant difference between real and sham stimuli after iTBS (p = 0.37). The greater hemodynamic changes suggest increased vasomotor reactivity after cTBS compared to iTBS. In addition, cTBS could decrease lateral inhibition, allowing activation of surrounding areas after cTBS.

Comprehensive assessment of the efficacy and safety of a clay mask in oily and acne skin.

BACKGROUND: Oily skin, characterized by excessive sebum production, can lead to acne and have psychosocial impacts due to changes in appearance. Recent research has shown interest in treatments for oil control, with kaolin and bentonite emerging as promising options. Despite their potential, comprehensive studies on these ingredients are still in the nascent stages. AIM: This study aimed to assess the efficacy of a clay mask (La Roche-Posay Effaclar Sebo-Controlling Mask) in reducing skin oiliness and acne, and its safety for use. METHODS: In this study, 75 adults with oily or combination skin were enrolled and provided with a clay mask for twice-weekly use over 4 weeks. Clinical assessments, using instruments like Sebumeter, Vapometer, and Corneometer, were conducted at baseline, and after 1, 2, and 4 weeks, evaluating acne lesions, skin irritation, sebum content, and skin hydration. Participant self-assessment questionnaires were also utilized for subjective evaluation. Statistical analyses were performed accordingly. RESULTS: The study revealed significant improvements in acne-related outcomes, sebum content, skin evenness, stratum corneum water content, and transepidermal water loss following the application of the clay mask. Pore area and porphyrin area showed no significant changes. Tolerance assessment showed reduced dryness and irritation, with self-assessment indicating high product acceptability and perceived oil control effectiveness. CONCLUSION: This study demonstrated the clay mask's efficacy in managing acne and oily skin, improving hydration and texture. Significant improvements in skin parameters and high product safety were observed, supporting its suitability.

Rationally designed multifunctional nanoparticles as GSH-responsive anticancer drug delivery systems based on host-guest polymers derived from dextran and ß-cyclodextrin.

Tumor proliferation and metastasis rely on energy provided by mitochondria. The hexokinase inhibitor lonidamine (LND) could suppress the activities in mitochondria, being a potential antitumor drug. However, limited water-solubility of LND may hinder its biomedical applications. Besides, the cancer-killing effect of LND is compromised by the high level of glutathione (GSH) in cancer cells. Therefore, it is urgent to find a proper method to simultaneously deliver LND and deplete GSH as well as monitor GSH level in cancer cells. Herein, a host polymer ß-cyclodextrin-polyethylenimine (ß-CD-PEI) and a guest polymer dextran-5-dithio-(2-nitrobenzoic acid) (Dextran-SS-TNB) were synthesized and allowed to form LND-loaded GSH-responsive nanoparticles through host-guest inclusion complexation between ß-CD and TNB as host and guest molecular moieties, respectively, which functioned as a system for simultaneous delivery of LND and -SS-TNB species into cancer cells. As a result, the delivery system could deplete GSH and elevate reactive oxygen species (ROS) level in cancer cells, further induce LND-based mitochondrial dysfunction and ROS-based immunogenic cell death (ICD), leading to a synergistic and efficient anticancer effect. In addition, -SS-TNB reacted with GSH to release TNB2-, which could be a probe with visible light absorption at 410 nm for monitoring the GSH level in the cells.

Biomass-based single- and double-network hydrogels derived from cellulose microfiber and chitosan for potential application as plant growing substrate.

A series of hydrogels were synthesized from renewable and low-cost micro-sized cellulose fiber. The single-network hydrogel was composed of cellulose fiber and a small amount of another polysaccharide, chitosan, which 'glued' individual cellulose fiber pieces together through Schiff-base bonding. The double-network hydrogel was constructed by adding a secondary network, the covalently crosslinked polyacrylamide, into the single-network hydrogel, which was synthesized by conducting Schiff-base reaction and free radical polymerization at the same time in a facile one-pot process. In both single- and double-network hydrogels, cellulose fiber constituted the dominant component. Both types of hydrogels exhibited good swelling properties. The double-network hydrogel showed much improved stability against soaking in water and higher salt tolerance. Germination experiment with choy sum seeds sowed on hydrogel surface showed that the seeds were able to germinate and further develop roots, shoots, and true leaves, demonstrating the potential of the biomass-derived hydrogels for soilless plant growing applications.

Functional identification of ZDS gene in apple (Malus halliana) and demonstration of it's role in improving saline-alkali stress tolerance.

Carotenoids are powerful antioxidants that mediate transfer of electrons, directly affect abiotic stress responses in plants through regulating activity of antioxidant enzymes. ζ-Carotene desaturase (ZDS) is a key enzyme in carotenoid biosynthesis pathway, which can catalyze ζ-carotene to form lycopene to regulate carotenoid biosynthesis and accumulation. However, the mechanism of its regulation of saline-alkali stress remains unclear. In this research, based on transcriptomic analysis of Malus halliana with a apple rootstock, we screened out ZDS gene (LOC103451012), with significantly high expression by saline-alkali stress, whose expression in the leaves was 10.8-fold than that of the control (0 h) under 48 h of stress. Subsequently, the MhZDS gene was isolated from M. halliana, and transgenic Arabidopsis thaliana, tobacco, and apple calli were successfully obtained through agrobacterium-mediated genetic transformation. We found that overexpression of MhZDS enhanced the tolerance of A. thaliana, tobacco and apple calli under saline-alkali stress and caused a variety of physiological and biochemical changes: compared with wild-type, transgenic plants grew better under saline stress and MhZDS-OE lines showed higher chlorophyll content, POD, SOD, CAT activities and proline content, lower electrical conductivity and MDA content. These results indicate that MhZDS plays an important role in plant resistance to saline-alkali stress, providing excellent resistance genes for the regulatory network of salinity stress response in apples and provide a theoretical basis for the breeding of apple varieties with strong saline-alkali resistance. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01333-5.

Effects of combined antibiotics on nitrification, bacteria and antibiotic resistance genes in activated sludge: Insights from legacy effect of antibiotics.

The effect of combined antibiotics exposure on nitrogen removal, microbial community assembly and proliferation of antibiotics resistance genes (ARGs) is a hotspot in activated sludge system. However, it is unclear that how the historical antibiotic stress affects the subsequent responses of microbes and ARGs to combined antibiotics. In this study, the effects of combined sulfamethoxazole (SMX) and trimethoprim (TMP) pollution on activated sludge under legacy of SMX or TMP stress with different doses (0.005-30 mg/L) were investigated to clarify antibiotic legacy effects. Nitrification activity was inhibited under higher level of combined exposure but a high total nitrogen removal (∼70%) occurred. Based on the full-scale classification, the legacy effect of past antibiotic stress had a marked effect on community composition of conditionally abundant taxa (CAT) and conditionally rare or abundant taxa (CRAT). Rare taxa (RT) were the keystone taxa in the microbial network, and the responses of hub genera were also affected by the legacy of antibiotic stress. Nitrifying bacteria and genes were inhibited by the antibiotics and aerobic denitrifying bacteria (Pseudomonas, Thaurea and Hydrogenophaga) were enriched under legacy of high dose, as were the key denitrifying genes (napA, nirK and norB). Furthermore, the occurrences and co-selection relationship of 94 ARGs were affected by legacy effect. While, some shared hosts (eg., Citrobacter) and hub ARGs (eg., mdtD, mdtE and acrD) were identified. Overall, antibiotic legacy could affect responses of activated sludge to combined antibiotic and the legacy effect was stronger at higher exposure levels.