Liquid Crystal Promoted Self-Assembly of Statistical Copolymers into Diverse Nanostructures with Precise Dimensions.

In both natural and synthetic systems, the segregation of multicomponent entities is vital for regulating functions and the ultimate usage of materials. To accomplish the desired properties via nanosegregation or microphase separation, great effort is usually demanded in the synthesis. For example, microphase-separated block copolymers rely on the delicate controlled/living polymerization of different monomers in sequence. Here, we demonstrate that a facile one-pot copolymerization can generate statistical side-chain copolymers exhibiting well-defined and diverse nanostructures. Two hemiphasmidic (or wedge-shaped) cyclooctene monomers were designed, differing in the peripheral tails of the wedges (dodecyl vs. tetraethylene glycol), with lengths of ca. 1 nm. When combining the two monomers together, the statistical copolymers can show columnar liquid crystal (LC) phase and microphase-separated structures of the two monomers, including sphere, cylinder, double gyroid, and lamella. To the best of our knowledge, this is the first time the gyroid phase has been achieved in statistical copolymers. We further demonstrate that changing the side chains to calamitic (or rod-like) mesogens or the backbone to less flexible polynorbornene, the statistical copolymers can also undergo microphase separation of the side chains. The intrinsic self-assembly scheme of statistical copolymers with mesogenic side chains, which are chemically accurate, affords the resultant nanostructures with precise periodicities at the 10- or sub-10-nm scale. Given the small chemical difference between the side-chain tails, microphase separation is promoted by the anisotropic packing of mesogens. It is validated that the statistical side-chain LC copolymers can be a versatile platform for creating nanostructured materials with tailored functionalities.

LN-439A, a novel BAP1 inhibitor, suppresses the growth of basal-like breast cancer by degrading KLF5.

Basal-like breast cancer (BLBC) is the most malignant subtype of breast cancer because of its aggressive clinical behaviour and lack of effective targeted agents. Krüppel-like factor 5 (KLF5) is an oncogenic transcription factor that is highly expressed in BLBC. The deubiquitinase (DUB) BRCA1-associated protein 1 (BAP1) stabilizes KLF5 and promotes BLBC growth and metastasis. Therefore, pharmacological inhibition of the BAP1‒KLF5 axis is an effective therapeutic strategy for BLBC. Here, through screening, we identified a series of tetrahydro-ß-carboline derivatives that effectively reduced the protein expression of KLF5 and exhibited strong antitumour activity. Among the investigated compounds, the lead compound LN-439A presented the strongest antitumour activity and inhibitory effect on KLF5 expression. LN-439A suppressed the proliferation and migration of BLBC cells, induced G2/M arrest, and induced apoptosis. Mechanistically, LN-439A functions as a small molecule catalytic inhibitor of BAP1 by binding to the catalytic pocket of BAP1, leading to the ubiquitination and degradation of KLF5. Consistent with this finding, the overexpression of KLF5 suppressed the antitumour effects of LN-439A. In summary, LN-439A is a promising therapeutic agent for BLBC that functions by targeting the BAP1‒KLF5 axis.

Loss of ZC4H2, an Arthrogryposis Multiplex Congenita Associated Gene, Promotes Osteoclastogenesis in Mice.

ZC4H2 encodes a C4H2-type zinc finger protein, mutations of which lead to a spectrum of diseases known as ZC4H2 associated rare disorders (ZARD). In addition to neurological phenotypes, the most typical symptoms of ZARD are multiple joint contractures of varying degrees, accompanied by abnormal development of muscles and bones, and osteoporosis in some cases. The pathogenic mechanisms of such bone related phenotypes, however, remain unclear. Here, we showed that ZC4H2 is expressed in the developing bones in mice. ZC4H2 knockout mice were neonatal-lethal and smaller in size, with reduced calcification of long bones. Upon induced loss of ZC4H2 postnatally, the femoral bones developed an osteoporosis-like phenotype, with reduced bone mineral density, bone-volume fraction, and trabecular bone number. Knockdown of ZC4H2 showed no clear effect on the expression of osteogenic differentiation genes in in vitro models using mesenchymal stem cells. Interestingly, ZC4H2 knockdown significantly enhanced osteoclast differentiation and bone resorption in induced bone marrow-derived macrophages. We further confirmed that the number of osteoclasts in the long bone of ZC4H2 knockout mice was increased, as well as the expression of the serum bone resorption/osteoporosis marker CTX-1. Our study unveils a new role of ZC4H2 in osteoclast differentiation and bone development, providing new clues on the pathology of ZARD.

Effectiveness and safety of adjuvant treatment of tislelizumab with or without chemotherapy in patients with high-risk upper tract urothelial carcinoma: a retrospective, real-world study.

BACKGROUND: Upper tract urothelial carcinoma (UTUC) is a rare subset of urothelial cancers with poor prognosis. No consensus exists on the benefit of adjuvant immunotherapy for patients with UTUCs after nephroureterectomy with curative intent and the existing studies are limited. Herein, this study aimed to evaluate the effectiveness and safety of adjuvant treatment of tislelizumab with or without chemotherapy in patients with high-risk UTUC. METHODS: A retrospective study was conducted on 63 patients with high-risk UTUC who received tislelizumab with or without gemcitabine-cisplatin (GC) chemotherapy regimen after surgery between January 2020 and December 2022. Data on demographic and clinical characteristics, surgical, outcomes, prognostic factors, and safety were collected and analyzed. RESULTS: Among the 63 patients with high-risk UTUC, the median age was 66 years (interquartile range 57-72), with 33 (52%) being male. The majority of patients with staged pT3 (44%) and pN0 (78%) disease. Fifty-one patients (81%) received tislelizumab plus GC chemotherapy, and 12 (19%) were treated with tislelizumab monotherapy. After the median follow-up of 26 months (range 1-47), 49 (78%) patients achieved stable disease. The 2-year disease-free survival (DFS) and 2-year overall survival were 78.68% (95% CI: 60.02-87.07%) and 81.40% (95% CI: 68.76-89.31%), respectively. The cycles of GC chemotherapy were independent prognostic factors for survival, with higher DFS (hazard ratio = 0.68, 95% CI, 0.50-0.93; p = 0.016) observed in the subgroup undergoing ≥ 3 cycles versus < 3 cycles of GC chemotherapy. Fifty-eight patients (92%) experienced at least one treatment-related adverse event (TRAE), with grade 3-4 TRAEs occurring in 13%. The most common grade 3-4 TRAEs were decreased white blood cells, thrombocytopenia, and ulcers. CONCLUSIONS: The study demonstrates promising clinical benefits and a manageable safety profile of the tislelizumab-based adjuvant regimen for patients with high-risk UTUC. This suggests that adjuvant immunotherapy represents a potential therapeutic strategy for this population.

Chloride-mediated enhancement in Cu(II)-catalyzed Fenton-like reaction: The overlooked reactive chlorine species.

The practical application of Cu(II)-catalyzed Fenton-like reaction (Cu(II)/H2O2) exhibits a low efficiency in the degradation of refractory compounds of wastewater. The impact of chloride ions (Cl-) on Fenton-like reactions have been investigated, but the influence mechanism is still unclear. Herein, the presence of Cl- (5 mM) significantly accelerated the degradation of benzoic acid (BA) under neutral conditions. The degradation of BA follows pseudo-first-order kinetics, with a degradation rate 7.3 times higher than the Cu(II)/H2O2 system. Multiple evidences strongly demonstrated that this reaction enables the production of reactive chlorine species (RCS) rather than HO• and high-valent copper (Cu(III)). The kinetic model revealed that Cl- could shift reactive species from the key intermediate (Cu(III)-chloro complexes) to RCS. Dichlorine radicals (Cl2•-) was discovered to play a crucial role in BA degradation, which was largely overlooked in previous reports. Although the reaction rate of Cl2•- with BA (k = 2.0 × 106 M-1 s-1) is lower than that of other species, its concentration is 10 orders of magnitude higher than that of Cu(III) and HO•. Furthermore, the exceptional efficacy of the Cu(II)/H2O2 system in BA degradation was observed in saline aquatic environments. This work sheds light on the previously unrecognized role of the metal-chloro complexes in production the RCS and water purification.

PI3K PROTAC overcomes the lapatinib resistance in PIK3CA-mutant HER2 positive breast cancer.

Although anti-HER2 therapy has made significant strides in reducing metastasis and relapse in HER2-positive breast cancer, resistance to agents like trastuzumab, pertuzumab, and lapatinib frequently develops in patients undergoing treatment. Previous studies suggest that the hyperactivation of the PI3K-AKT signaling pathway by PIK3CA/PTEN gene mutations is implicated in HER2 resistance. In this study, we introduce a novel PI3K-p110α Proteolysis TAargeting Chimera (PROTAC) that effectively inhibits the proliferation of breast cancer cells by degrading PI3K-p110α. When tested in two lapatinib-resistant cell lines, JIMT1 and MDA-MB-453, both of which harbor PIK3CA mutations, the PI3K PROTAC notably reduced cell proliferation and induced G1 phase cell cycle arrest. Importantly, even at very low concentrations, PI3K PROTAC restored sensitivity to lapatinib. Furthermore, the efficacy of PI3K PROTAC surpassed that of Alpelisib, a selective PI3K-p110α kinase inhibitor in clinic. The superior performance of PI3K PROTAC was also confirmed in lapatinib-resistant breast cancer xenograft tumors and patient-derived breast cancer organoids (PDOs). In conclusion, this study reveals that the novel PI3K PROTAC we synthesized could serve as an effective agent to overcome lapatinib resistance.

The Application of Multi-Parameter Multi-Modal Technology Integrating Biological Sensors and Artificial Intelligence in the Rapid Detection of Food Contaminants.

Against the backdrop of continuous socio-economic development, there is a growing concern among people about food quality and safety. Individuals are increasingly realizing the critical importance of healthy eating for bodily health; hence the continuous rise in demand for detecting food pollution. Simultaneously, the rapid expansion of global food trade has made people's pursuit of high-quality food more urgent. However, traditional methods of food analysis have certain limitations, mainly manifested in the high degree of reliance on personal subjective judgment for assessing food quality. In this context, the emergence of artificial intelligence and biosensors has provided new possibilities for the evaluation of food quality. This paper proposes a comprehensive approach that involves aggregating data relevant to food quality indices and developing corresponding evaluation models to highlight the effectiveness and comprehensiveness of artificial intelligence and biosensors in food quality evaluation. The potential prospects and challenges of this method in the field of food safety are comprehensively discussed, aiming to provide valuable references for future research and practice.

Bilateral gene therapy in children with autosomal recessive deafness 9: single-arm trial results.

Gene therapy is a promising approach for hereditary deafness. We recently showed that unilateral AAV1-hOTOF gene therapy with dual adeno-associated virus (AAV) serotype 1 carrying human OTOF transgene is safe and associated with functional improvements in patients with autosomal recessive deafness 9 (DFNB9). The protocol was subsequently amended and approved to allow bilateral gene therapy administration. Here we report an interim analysis of the single-arm trial investigating the safety and efficacy of binaural therapy in five pediatric patients with DFNB9. The primary endpoint was dose-limiting toxicity at 6 weeks, and the secondary endpoint included safety (adverse events) and efficacy (auditory function and speech perception). No dose-limiting toxicity or serious adverse event occurred. A total of 36 adverse events occurred. The most common adverse events were increased lymphocyte counts (6 out of 36) and increased cholesterol levels (6 out of 36). All patients had bilateral hearing restoration. The average auditory brainstem response threshold in the right (left) ear was >95 dB (>95 dB) in all patients at baseline, and the average auditory brainstem response threshold in the right (left) ear was restored to 58 dB (58 dB) in patient 1, 75 dB (85 dB) in patient 2, 55 dB (50 dB) in patient 3 at 26 weeks, and 75 dB (78 dB) in patient 4 and 63 dB (63 dB) in patient 5 at 13 weeks. The speech perception and the capability of sound source localization were restored in all five patients. These results provide preliminary insights on the safety and efficacy of binaural AAV gene therapy for hereditary deafness. The trial is ongoing with longer follow-up to confirm the safety and efficacy findings. Chinese Clinical Trial Registry registration: ChiCTR2200063181 .

Engineering of the AAV-Compatible Hair Cell-Specific Small-Size Myo15 Promoter for Gene Therapy in the Inner Ear.

Adeno-associated virus (AAV)-mediated gene therapy is widely applied to treat numerous hereditary diseases in animal models and humans. The specific expression of AAV-delivered transgenes driven by cell type-specific promoters should further increase the safety of gene therapy. However, current methods for screening cell type-specific promoters are labor-intensive and time-consuming. Herein, we designed a "multiple vectors in one AAV" strategy for promoter construction in vivo. Through this strategy, we truncated a native promoter for Myo15 expression in hair cells (HCs) in the inner ear, from 1,611 bp down to 1,157 bp, and further down to 956 bp. Under the control of these 2 promoters, green fluorescent protein packaged in AAV-PHP.eB was exclusively expressed in the HCs. The transcription initiation ability of the 2 promoters was further verified by intein-mediated otoferlin recombination in a dual-AAV therapeutic system. Driven by these 2 promoters, human otoferlin was selectively expressed in HCs, resulting in the restoration of hearing in treated Otof -/- mice for at least 52 weeks. In summary, we developed an efficient screening strategy for cell type-specific promoter engineering and created 2 truncated Myo15 promoters that not only restored hereditary deafness in animal models but also show great potential for treating human patients in future.

Predicting the Posture of High-Rise Building Machines Based on Multivariate Time Series Neural Network Models.

High-rise building machines (HBMs) play a critical role in the successful construction of super-high skyscrapers, providing essential support and ensuring safety. The HBM's climbing system relies on a jacking mechanism consisting of several independent jacking cylinders. A reliable control system is imperative to maintain the smooth posture of the construction steel platform (SP) under the action of the jacking mechanism. Long Short-Term Memory (LSTM), Gated Recurrent Unit (GRU), and Temporal Convolutional Network (TCN) are three multivariate time series (MTS) neural network models that are used in this study to predict the posture of HBMs. The models take pressure and stroke measurements from the jacking cylinders as inputs, and their outputs determine the levelness of the SP and the posture of the HBM at various climbing stages. The development and training of these neural networks are based on historical on-site data, with the predictions subjected to thorough comparative analysis. The proposed LSTM and GRU prediction models have similar performances in the prediction process of HBM posture, with medians R2 of 0.903 and 0.871, respectively. However, the median MAE of the GRU prediction model is more petite at 0.4, which exhibits stronger robustness. Additionally, sensitivity analysis showed that the change in the levelness of the position of the SP portion of the HBM exhibited high sensitivity to the stroke and pressure of the jacking cylinder, which clarified the position of the cylinder for adjusting the posture of the HBM. The results show that the MTS neural network-based prediction model can change the HBM posture and improve work stability by adjusting the jacking cylinder pressure value of the HBM.

Using UV/peracetic acid as pretreatment for subsequent bio-treatment of antibiotic-containing wastewater treatment: Mitigating microbial inhibition and antibiotic resistance genes proliferation.

UV/peracetic acid (PAA) treatment presents a promising approach for antibiotic removal, but its effects on microbial community and proliferation of antibiotic resistance genes (ARGs) during the subsequent bio-treatment remain unclear. Thus, we evaluated the effects of the UV/PAA on tetracycline (TTC) degradation, followed by introduction of the treated wastewater into the bio-treatment system to monitor changes in ARG expression and biodegradability. Results demonstrated effective TTC elimination by the UV/PAA system, with carbon-centered radicals playing a significant role. Crucially, the UV/PAA system not only eliminated antibacterial activity but also inhibited potential ARG host growth, thereby minimizing the emergence and dissemination of ARGs during subsequent bio-treatment. Additionally, the UV/PAA system efficiently removed multi-antibiotic resistant bacteria and ARGs from the bio-treatment effluent, preventing ARGs from being released into the environment. Hence, we propose a multi-barrier strategy for treating antibiotic-containing wastewater, integrating UV/PAA pre-treatment and post-disinfection with bio-treatment. The inhibition of ARGs transmission by the integrated system was verified through actual soil testing, confirming its effectiveness in preventing ARGs dissemination in the surrounding natural ecosystem. Overall, the UV/PAA treatment system offers a promising solution for tackling ARGs challenges by controlling ARGs proliferation at the source and minimizing their release at the end of the treatment process.

Toxoflavin analog D43 exerts antiproliferative effects on breast cancer by inducing ROS-mediated apoptosis and DNA damage.

Triple-negative breast cancer (TNBC) is regarded as the deadliest subtype of breast cancer because of its high heterogeneity, aggressiveness, and limited treatment options. Toxoflavin has been reported to possess antitumor activity. In this study, a series of toxoflavin analogs were synthesized, among which D43 displayed a significant dose-dependent inhibitory effect on the proliferation of TNBC cells (MDA-MB-231 and HCC1806). Additionally, D43 inhibited DNA synthesis in TNBC cells, leading to cell cycle arrest at the G2/M phase. Furthermore, D43 consistently promoted intracellular ROS generation, induced DNA damage, and resulted in apoptosis in TNBC cells. These effects could be reversed by N-acetylcysteine. Moreover, D43 significantly inhibited the growth of breast cancer patient-derived organoids and xenografts with a favorable biosafety profile. In conclusion, D43 is a potent anticancer agent that elicits significant antiproliferation, oxidative stress, apoptosis, and DNA damage effects in TNBC cells, and D43 holds promise as a potential candidate for the treatment of TNBC.

Electrochemically Exfoliated Graphene Additive-Free Inks for 3D Printing Customizable Monolithic Integrated Micro-Supercapacitors on a Large Scale.

Three-dimensional (3D) printing technology with enhanced fidelity can achieve multiple functionalities and boost electrochemical performance of customizable planar micro-supercapacitors (MSCs), however, precise structural control of additive-free graphene-based macro-assembly electrode for monolithic integrated MSCs (MIMSCs) remains challenging. Here, the large-scale 3D printing fabrication of customizable planar MIMSCs is reported utilizing additive-free, high-quality electrochemically exfoliated graphene inks, which is not required the conventional cryogenic assistance during the printing process and any post-processing reduction. The resulting MSCs reveal an extremely small engineering footprint of 0.025 cm2, exceptionally high areal capacitance of 4900 mF cm-2, volumetric capacitance of 195.6 F cm-3, areal energy density of 2.1 mWh cm-2, and unprecedented volumetric energy density of 23 mWh cm-3 for a single cell, surpassing most previously reported 3D printed MSCs. The 3D printed MIMSC pack is further demonstrated, with the maximum areal cell count density of 16 cell cm-2, the highest output voltage of 192.5 V and the largest output voltage per unit area of 56 V cm-2 up to date are achieved. This work presents an innovative solution for processing high-performance additive-free graphene ink and realizing the large-scale production of 3D printed MIMSCs for planar energy storage.

Hair cell-specific Myo15 promoter-mediated gene therapy rescues hearing in DFNB9 mouse model.

Adeno-associated viral (AAV) vectors are increasingly used as vehicles for gene delivery to treat hearing loss. However, lack of specificity of the transgene expression may lead to overexpression of the transgene in nontarget tissues. In this study, we evaluated the expression efficiency and specificity of transgene delivered by AAV-PHP.eB under the inner ear sensory cell-specific Myo15 promoter. Compared with the ubiquitous CAG promoter, the Myo15 promoter initiates efficient expression of the GFP fluorescence reporter in hair cells, while minimizing non-specific expression in other cell types of the inner ear and CNS. Furthermore, using the Myo15 promoter, we constructed an AAV-mediated therapeutic system with the coding sequence of OTOF gene. After inner ear injection, we observed apparent hearing recovery in Otof-/- mice, highly efficient expression of exogenous otoferlin, and significant improvement in the exocytosis function of inner hair cells. Overall, our results indicate that gene therapy mediated by the hair cell-specific Myo15 promoter has potential clinical application for the treatment of autosomal recessive deafness and yet for other hereditary hearing loss related to dysfunction of hair cells.

AAV1-hOTOF gene therapy for autosomal recessive deafness 9: a single-arm trial.

BACKGROUND: Autosomal recessive deafness 9, caused by mutations of the OTOF gene, is characterised by congenital or prelingual, severe-to-complete, bilateral hearing loss. However, no pharmacological treatment is currently available for congenital deafness. In this Article, we report the safety and efficacy of gene therapy with an adeno-associated virus (AAV) serotype 1 carrying a human OTOF transgene (AAV1-hOTOF) as a treatment for children with autosomal recessive deafness 9. METHODS: This single-arm, single-centre trial enrolled children (aged 1-18 years) with severe-to-complete hearing loss and confirmed mutations in both alleles of OTOF, and without bilateral cochlear implants. A single injection of AAV1-hOTOF was administered into the cochlea through the round window. The primary endpoint was dose-limiting toxicity at 6 weeks after injection. Auditory function and speech were assessed by appropriate auditory perception evaluation tools. All analyses were done according to the intention-to-treat principle. This trial is registered with Chinese Clinical Trial Registry, ChiCTR2200063181, and is ongoing. FINDINGS: Between Oct 19, 2022, and June 9, 2023, we screened 425 participants for eligibility and enrolled six children for AAV1-hOTOF gene therapy (one received a dose of 9 × 1011 vector genomes [vg] and five received 1·5 × 1012 vg). All participants completed follow-up visits up to week 26. No dose-limiting toxicity or serious adverse events occurred. In total, 48 adverse events were observed; 46 (96%) were grade 1-2 and two (4%) were grade 3 (decreased neutrophil count in one participant). Five children had hearing recovery, shown by a 40-57 dB reduction in the average auditory brainstem response (ABR) thresholds at 0·5-4·0 kHz. In the participant who received the 9 × 1011 vg dose, the average ABR threshold was improved from greater than 95 dB at baseline to 68 dB at 4 weeks, 53 dB at 13 weeks, and 45 dB at 26 weeks. In those who received 1·5 × 1012 AAV1-hOTOF, the average ABR thresholds changed from greater than 95 dB at baseline to 48 dB, 38 dB, 40 dB, and 55 dB in four children with hearing recovery at 26 weeks. Speech perception was improved in participants who had hearing recovery. INTERPRETATION: AAV1-hOTOF gene therapy is safe and efficacious as a novel treatment for children with autosomal recessive deafness 9. FUNDING: National Natural Science Foundation of China, National Key R&D Program of China, Science and Technology Commission of Shanghai Municipality, and Shanghai Refreshgene Therapeutics.

Thrombospondin 2 is a novel biomarker of essential hypertension and associated with nocturnal Na+ excretion and insulin resistance.

BACKGROUND: Thrombospondins (TSPs) play important roles in several cardiovascular diseases. However, the association between circulating (plasma) thrombospondin 2 (TSP2) and essential hypertension remains unclear. The present study was aimed to investigate the association of circulating TSP2 with blood pressure and nocturnal urine Na+ excretion and evaluate the predictive value of circulating TSP2 in subjects with hypertension. METHODS AND RESULTS: 603 newly diagnosed essential hypertensive subjects and 508 healthy subjects were preliminarily screened, 47 healthy subjects and 40 newly diagnosed essential hypertensive subjects without any chronic diseases were recruited. The results showed that the levels of circulating TSP2 were elevated in essential hypertensive subjects. The levels of TSP2 positively associated with systolic blood pressure (SBP), diastolic blood pressure (DBP), and other clinical parameters, including homeostasis model assessment of insulin resistance (HOMA-IR), brachial-ankle pulse wave velocity, and serum triglycerides, but negatively associated with nocturnal urine Na+ concentration and excretion and high-density lipoprotein cholesterol. Results of multiple linear regressions showed that HOMA-IR and nocturnal Na+ excretion were independent factors related to circulating TSP2. Mantel-Haenszel chi-square test displayed linear relationships between TSP2 and SBP (χ2 = 35.737) and DBP (χ2 = 26.652). The area under receiver operating characteristic curve (AUROC) of hypertension prediction was 0.901. CONCLUSION: Our study suggests for the first time that the circulating levels of TSP2 may be a novel potential biomarker for essential hypertension. The association between TSP2 and blood pressure may be, at least in part, related to the regulation of renal Na+ excretion, insulin resistance, and/or endothelial function.

Preclinical evaluation of the efficacy and safety of AAV1-hOTOF in mice and nonhuman primates.

Pathogenic mutations in the OTOF gene cause autosomal recessive hearing loss (DFNB9), one of the most common forms of auditory neuropathy. There is no biological treatment for DFNB9. Here, we designed an OTOF gene therapy agent by dual-adeno-associated virus 1 (AAV1) carrying human OTOF coding sequences with the expression driven by the hair cell-specific promoter Myo15, AAV1-hOTOF. To develop a clinical application of AAV1-hOTOF gene therapy, we evaluated its efficacy and safety in animal models using pharmacodynamics, behavior, and histopathology. AAV1-hOTOF inner ear delivery significantly improved hearing in Otof-/- mice without affecting normal hearing in wild-type mice. AAV1 was predominately distributed to the cochlea, although it was detected in other organs such as the CNS and the liver, and no obvious toxic effects of AAV1-hOTOF were observed in mice. To further evaluate the safety of Myo15 promoter-driven AAV1-transgene, AAV1-GFP was delivered into the inner ear of Macaca fascicularis via the round window membrane. AAV1-GFP transduced 60%-94% of the inner hair cells along the cochlear turns. AAV1-GFP was detected in isolated organs and no significant adverse effects were detected. These results suggest that AAV1-hOTOF is well tolerated and effective in animals, providing critical support for its clinical translation.

Computer Vision Interaction Design in Sustainable Urban Development: A Case Study of Roof Garden Landscape Plants in Marine Cities.

The rapid urbanization and the increasing need for sustainable development have led to the emergence of green roof landscapes in ocean cities. These rooftop gardens provide numerous environmental benefits and contribute to the overall well-being of urban dwellers. However, optimizing the design and interaction experience of green roof landscapes requires the integration of intelligent technologies. This paper explores the application of computer visual design techniques, specifically 3DMAX modeling and virtual reality, in the intelligent interaction design of green roof landscape plants in ocean cities. Designers can use computer visual design (3DMAX) and other technologies to interact intelligently with the roof landscape in order to improve landscape design. Through case studies, this indicated that computer vision is excellent for image processing of rooftop landscapes and also demonstrates a high degree of compatibility between computer vision and green rooftop landscape plant design in marine cities. This paper highlights the significance of intelligent interaction design and computer visual design techniques in optimizing the integration of green roof landscape plants in ocean cities. It emphasizes the potential of 3DMAX modeling and VR technology in creating immersive and engaging experiences for designers, users, and stakeholders alike. The findings contribute to the growing body of knowledge in the field of sustainable urban development and provide insights for designers, policymakers, and researchers seeking to enhance green roof landscapes in ocean cities. The dissertation highlights the potential of using computer vision design techniques to enhance the roof garden landscaping process and advocates for more efficient and effective ways to design, visualize, and improve rooftop gardens by utilizing software equipped with computer vision technology such as 3DMAX, ultimately contributing to the advancement of sustainable urban landscapes.

Phosphorus recovery from sewage sludge ash (SSA): An integrated technical, environmental and economic assessment of wet-chemical and thermochemical methods.

Incineration is a promising disposal method for sewage sludge (SS), enriching more than 90% of phosphorus (P) in the influent into the powdered product, sewage sludge ash (SSA), which is convenient for further P recovery. Due to insufficient bioavailable P and enriched heavy metals (HMs) in SSA, it is limited to be used directly as fertilizer. Hence, this paper provides an overview of P transformation in SS incineration, characterization of SSA components, and wet-chemical and thermochemical processes for P recovery with a comprehensive technical, economic, and environmental assessment. P extraction and purification is an important technical step to achieve P recovery from SSA, where the key to all technologies is how to achieve efficient separation of P and HMs at a low economic and environmental cost. It can be clear seen from the review that the economics of P recovery from SSA are often weak due to many factors. For example, the cost of wet-chemical methods is approximately 5∼6 €/kg P, while the cost of recovering P by thermochemical methods is about 2∼3 €/kg P, which is slightly higher than the current P fertilizer (1 €/kg P). So, for now, legislation is significant for promoting P recovery from SSA. In this regard, the relevant experience in Europe is worth learning from countries that have not yet carried out P recovery from SSA, and to develop appropriate policies and legislation according to their own national conditions.

Peracetic acid disinfection induces antibiotic-resistant E. coli into VBNC state but ineffectively eliminates the transmission potential of ARGs.

The occurrence of a viable but nonculturable (VBNC) state in antibiotic-resistant E. coli (AR E. coli) and inefficient degradation of their antibiotic resistance genes (ARGs) may cause potential health risks during disinfection. Peracetic acid (PAA) is an alternative disinfectant for replacing chlorine-based oxidants in wastewater treatment, and the potential of PAA to induce a VBNC state in AR E. coli and to remove the transformation functionality of ARGs were investigated for the first time. Results show that PAA exhibits excellent performance in inactivating AR E. coli (over 7.0-logs) and persistently inhibiting its regeneration. After PAA disinfection, insignificant changes in the ratio of living to dead cells (∼4%) and the level of cell metabolism, indicating that AR E. coli were induced into VBNC states. Unexpectedly, PAA was found to induce AR E. coli into VBNC state by destroying the proteins containing reactive amino acids at thiol, thioether and imidazole groups, rather than the result of membrane damage, oxidative stress, lipid destruction and DNA disruption in the conventional disinfection processes. Moreover, the result of poor reactivity between PAA and plasmid strands and bases confirmed that PAA hardly reduced the abundance of ARGs and damaged the plasmid's integrity. Transformation assays and real environment validation indicated that PAA-treated AR E. coli could release large abundance of naked ARGs with high-efficiency transformation functionality (∼5.4 × 10-4 - ∼8.3 × 10-6) into the environment. This study has significant environmental implications for assessing the transmission of antimicrobial resistance during PAA disinfection.