De novo protein design with a denoising diffusion network independent of pretrained structure prediction models.

The recent success of RFdiffusion, a method for protein structure design with a denoising diffusion probabilistic model, has relied on fine-tuning the RoseTTAFold structure prediction network for protein backbone denoising. Here, we introduce SCUBA-diffusion (SCUBA-D), a protein backbone denoising diffusion probabilistic model freshly trained by considering co-diffusion of sequence representation to enhance model regularization and adversarial losses to minimize data-out-of-distribution errors. While matching the performance of the pretrained RoseTTAFold-based RFdiffusion in generating experimentally realizable protein structures, SCUBA-D readily generates protein structures with not-yet-observed overall folds that are different from those predictable with RoseTTAFold. The accuracy of SCUBA-D was confirmed by the X-ray structures of 16 designed proteins and a protein complex, and by experiments validating designed heme-binding proteins and Ras-binding proteins. Our work shows that deep generative models of images or texts can be fruitfully extended to complex physical objects like protein structures by addressing outstanding issues such as the data-out-of-distribution errors.

Synergistic Procoagulant Mechanism and Application of Kaolin-Zeolite Composite Hemostat for Effective Hemorrhage Control.

Achieving timely and effective hemorrhage control is imperative for the survival of individuals with severe bleeding. Hemostatic materials, by enhancing the natural cell-based coagulation response, are essential tools in modern and military medical practice for controlling bleeding, especially in emergency and surgical settings. Here, we report a new type of composite hemostatic material with two different aluminosilicate-based components, kaolin and zeolite, which synergistically work together in different stages of the coagulation cascade reactions. Kaolin can effectively activate the clotting factor FXII in the early stage, and zeolite can accumulate and assemble FXa and FVa on its surface and thereafter lead to the formation of highly active thrombin in the later stage. The synergistic action mechanism between kaolin and zeolite significantly boosts the levels of FXIIa and FXa, and it also greatly enhances plateau thrombin activity. For practical application, a kaolin-modified zeolite gauze is fabricated, and it demonstrates excellent hemostatic effectiveness. Compared to the combat gauze currently used in front-line treatment, it reduces blood loss by 75% and shortens hemostasis time by 33% in a rabbit femoral artery injury model. In addition, this kaolin-zeolite gauze has no heat release problem and a nearly zero particle shedding rate, which greatly decreases the safety risk compared to current commercial inorganic-based hemostatic gauzes.

A near-complete chromosome-level genome assembly of looseleaf lettuce (Lactuca sativa var. crispa).

Lettuce (Lactuca sativa L., Asteraceae) is one of the most important vegetable crops, known for its various horticultural types and significant morphological variation. The first reference genome of lettuce, a crisphead type (L. sativa var. capitata cv. Salinas), was previously released. Here, we reported a near-complete chromosome-level reference genome for looseleaf lettuce (L. sativa var. crispa). PacBio high-fidelity sequencing, Oxford Nanopore, and Hi-C technologies were employed to produce genome assembly. The final assembly is 2.59 Gb in length with a contig N50 of 205.47 Mb, anchored onto nine chromosomes, containing 14 recognizable telomeres and only 11 gaps. Repetitive sequences account for 77.11% of the genome, and 41,375 protein-coding genes were predicted, with 99.10% of these assigned functional annotations. This chromosome-level genome enriched genomic resources for various horticultural types of lettuce and will facilitate the characterization of morphological variation and genetic improvement in lettuce.

Transverse Spin Selectivity in Helical Nanofibers Prepared without Any Chiral Molecule.

In the last decade, chirality-induced spin selectivity (CISS) has undergone intensive study. However, there remain several critical issues, such as the microscopic mechanism of CISS, especially transverse CISS where electrons are injected perpendicular to the helix axis of chiral molecules, quantitative agreement between experiments and theory, and at which level the molecular handedness is key to the CISS. Here, we address these issues by performing a combined experimental and theoretical study on conducting polyaniline helical nanofibers which are synthesized in the absence of any chiral species. Large spin polarization is measured in both left- and right-handed nanofibers for electrons injected perpendicular to their helix axis, and it will be reversed by switching the nanofiber handedness. We first develop a theoretical model to study this transverse CISS and quantitatively explain the experiment. Our results reveal that our theory provides a unifying scheme to interpret a number of CISS experiments, quantitative agreement between experiments and numerical calculations can be achieved by weak spin-orbit coupling, and the supramolecular handedness is sufficient for spin selectivity without any chiral species.

Exploring Prostate Cancer Incidence Trends and Age Change in Cancer Registration Areas of Jiangsu Province, China, 2009 to 2019.

(1) Background: Over the past few decades, Jiangsu Province, China, has witnessed a remarkable surge in the incidence of prostate cancer (PCa), accompanied by notable demographic shifts; (2) Methods: PCa data for Jiangsu Province from 2009 to 2019 were obtained from the Jiangsu Cancer Registry. We calculated crude and age-specific incidence rates (ASIRs), average age at onset, and age-specific composition ratios. Standardization was performed based on the Segi's world population. Birth cohorts (1929-2019) were analyzed to assess PCa incidence by birth year. Trend analysis was conducted using the Joinpoint Regression Model, and average annual percent changes (AAPCs) with corresponding 95% confidence interval (CI) were computed. A linear regression model was used to analyze the relationship between the average age at diagnosis and calendar years; (3) Results: The ASIRs of PCa in Jiangsu Province increased significantly, with an AAPC of 11.25% (95%CI: 10.09%, 12.42%) from 2009 to 2019. This increase was observed across all age groups, particularly among those aged 0-59 years. Birth cohort analysis revealed a rising trend with earlier birth years showing higher incidence, notably in the 1959 cohort. In rural areas, the age-standardized average age at onset of PCa decreased from 75.45 years in 2009 to 73.39 years in 2019, and the peak age group shifted from 75-79 years in 2009 to 70-74 years in 2019; (4) Conclusions: The rising incidence of PCa in Jiangsu Province, along with an increased proportion of cases in younger age groups, highlights the need for targeted interventions.

Nanoparticles prepared with biotin-esterified debranched starch as an oral carrier to improve the stability and antioxidant activity of resveratrol.

In this study, biotin esterified debranched starch (Bio-DBS) nanoparticles with different molecular weights were prepared to improve the stability and antioxidant activity of resveratrol. The molecular weights of branched starch (DBS3, DBS9 and DBSp) determined by high-performance size-exclusion chromatography (HPSEC) were 3306, 3696, and 4688, respectively. Biotin was covalently coupled to DBS through the esterification reaction as a new material to prepare nanoparticles. The morphology, particle size, and loading capacity of Bio-DBS nanoparticles were all related to the molecular weights of DBS. The 1H NMR results indicated that there was a hydrogen bonding interaction between Bio-DBS and resveratrol, which contributed to the photochemical and antioxidant activity of resveratrol in the nanoparticles. The highest encapsulation efficiency (78.9 %) and loading capacity (15.78 %) of resveratrol were observed in Bio-DBS3 nanoparticles. Additionally, the cell viability was over 80 % when the concentration of Bio-DBS3 reached to 200 µg/mL. The Bio-DBS nanoparticles significantly improved the thermal stability, photostability, and antioxidant properties of resveratrol. Therefore, the Bio-DBS nanoparticles prepared in this study can be used as a promising carrier to improve the stability and antioxidant activity of resveratrol and may have potential applications in oral delivery.

Integrated environmental assessment of a diversion-project-type urban water source considering the risks of novel and legacy contaminants.

The water diversion project is an effective engineering approach to overcome water scarcity as a water source for the area. However, the complex environmental conditions of long-distance water diversion bring many uncertainties for water security. In this study, we assessed the pollution condition and risk levels of emerging contaminants and traditional contaminants in the water and soil along a water diversion project in Tianjin. Then, we assessed the influence of eco-economic characteristics on environmental conditions and established a comprehensive assessment framework of water source sustainability by analytic hierarchy process (AHP). The results showed that excessive nutrient elements and heavy metal pollution mainly contributed to environmental problems in the water source area. Contrary to pollution assessment, the soil ecosystem was more subject to environmental pressure due to atmospheric deposition. The health risk assessment indicated that all contaminants had negligible non-carcinogenic risks for adults, with arsenic being considered a priority pollutant. The statistical analysis results indicated land use allocation was the most important factor in the environmental management of the water source area. According to the result of the integrated environmental assessment, the main characteristics of pressure zones were high pollution levels and human activity intensity. It is urgent to control agricultural pollution and allocate land use rationally for water source pressure zones. By considering the risks of traditional and emerging contaminants in water and soil, this study could support urban water source management and the sustainable development of the water diversion project.

Luciferase-Loaded Calcium Phosphate Nanoparticles for Persistent Bioluminescence Imaging of Orthotopic Breast Tumors.

Bioluminescence imaging (BLI) is an important noninvasive optical imaging technique that has been widely used to monitor many biological processes due to its high sensitivity, resolution, and signal-to-noise ratio. However, the BLI technique based on the firefly luciferin-luciferase system is limited by the expression of exogenous luciferase and the short half-life of firefly luciferin, which pose challenges for long-term tracking in vivo. To solve the problems, here we rationally designed an intelligent strategy for persistent BLI in tumors by combining luciferase-loaded calcium phosphate nanoparticles (Luc@CaP NPs) to provide luciferase and the probe Cys(SEt)-Lys-CBT (CKCBT) to slowly produce the luciferase substrate amino luciferin (Am-luciferin). Luc@CaP NPs constructed with CaP as a carrier could enable luciferase activity to be maintained in vivo for at least 12 h. And compared to the conventional substrate luciferin, CKCBT apparently prolonged the BL time by up to 2 h through GSH-induced intracellular self-assembly and subsequent protease degradation-induced release of Am-luciferin. We anticipate that this strategy could be applied for clinical translation in more disease diagnosis and treatment in the near future.

The IL-33/ST2 Axis Protects Retinal Ganglion Cells by Modulating the Astrocyte Response After Optic Nerve Injury.

IL-33 and its receptor ST2 play crucial roles in tissue repair and homeostasis. However, their involvement in optic neuropathy due to trauma and glaucoma remains unclear. Here, we report that IL-33 and ST2 were highly expressed in the mouse optic nerve and retina. Deletion of IL-33 or ST2 exacerbated retinal ganglion cell (RGC) loss, retinal thinning, and nerve fiber degeneration following optic nerve (ON) injury. This heightened retinal neurodegeneration correlated with increased neurotoxic astrocytes in Il33-/- mice. In vitro, rIL-33 mitigated the neurotoxic astrocyte phenotype and reduced the expression of pro-inflammatory factors, thereby alleviating the RGC death induced by neurotoxic astrocyte-conditioned medium in retinal explants. Exogenous IL-33 treatment improved RGC survival in Il33-/- and WT mice after ON injury, but not in ST2-/- mice. Our findings highlight the role of the IL-33/ST2 axis in modulating reactive astrocyte function and providing neuroprotection for RGCs following ON injury.

Investigating the impact of extreme weather events and related indicators on cardiometabolic multimorbidity.

BACKGROUND: The impact of weather on human health has been proven, but the impact of extreme weather events on cardiometabolic multimorbidity (CMM) needs to be urgently explored. OBJECTIVES: Investigating the impact of extreme temperature, relative humidity (RH), and laboratory testing parameters at admission on adverse events in CMM hospitalizations. DESIGNS: Time-stratified case-crossover design. METHODS: A distributional lag nonlinear model with a time-stratified case-crossover design was used to explore the nonlinear lagged association between environmental factors and CMM. Subsequently, unbalanced data were processed by 1:2 propensity score matching (PSM) and conditional logistic regression was employed to analyze the association between laboratory indicators and unplanned readmissions for CMM. Finally, the previously identified environmental factors and relevant laboratory indicators were incorporated into different machine learning models to predict the risk of unplanned readmission for CMM. RESULTS: There are nonlinear associations and hysteresis effects between temperature, RH and hospital admissions for a variety of CMM. In addition, the risk of admission is higher under low temperature and high RH conditions with the addition of particulate matter (PM, PM2.5 and PM10) and O3_8h. The risk is greater for females and adults aged 65 and older. Compared with first quartile (Q1), the fourth quartile (Q4) had a higher association between serum calcium (HR = 1.3632, 95% CI: 1.0732 ~ 1.7334), serum creatinine (HR = 1.7987, 95% CI: 1.3528 ~ 2.3958), fasting plasma glucose (HR = 1.2579, 95% CI: 1.0839 ~ 1.4770), aspartate aminotransferase/ alanine aminotransferase ratio (HR = 2.3131, 95% CI: 1.9844 ~ 2.6418), alanine aminotransferase (HR = 1.7687, 95% CI: 1.2388 ~ 2.2986), and gamma-glutamyltransferase (HR = 1.4951, 95% CI: 1.2551 ~ 1.7351) were independently and positively associated with unplanned readmission for CMM. However, serum total bilirubin and High-Density Lipoprotein (HDL) showed negative correlations. After incorporating environmental factors and their lagged terms, eXtreme Gradient Boosting (XGBoost) demonstrated a more prominent predictive performance for unplanned readmission of CMM patients, with an average area under the receiver operating characteristic curve (AUC) of 0.767 (95% CI:0.7486 ~ 0.7854). CONCLUSIONS: Extreme cold or wet weather is linked to worsened adverse health effects in female patients with CMM and in individuals aged 65 years and older. Moreover, meteorologic factors and environmental pollutants may elevate the likelihood of unplanned readmissions for CMM.

Comparison of novel and traditional anthropometric indices in Eastern-China adults: which is the best indicator of the metabolically obese normal weight phenotype?

BACKGROUND: People with the metabolically obese normal weight (MONW) phenotype have been confirmed to significantly increase the risk of unfavorable health consequences. This study aimed to investigate the relationships between traditional and novel anthropometric indices with the MONW phenotype and compare the predictive ability of different anthropometric indices in identifying individuals with the MONW phenotype. METHODS: This cross-sectional study involved a total of 26,332 participants aged 18 years or older with a normal weight from Nanjing, China. Sociodemographic information, biochemical measurements, and anthropometric indices were collected. The novel body fat anthropometric indices included body shape index (ABSI), body roundness index (BRI), abdominal volume index (AVI), weight-adjusted-waist index (WWI), body adiposity index (BAI), conicity index (CI), waist-hip-height ratio (WHHR), as well as traditional indices such as waist circumference (WC), hip circumference (HC), body mass index (BMI), waist-to-hip ratio (WHR), and waist-to-height ratio (WHtR).The prevalence ratio (PR) from modified poisson regression and area under the receiver-operating characteristic curve (AUC) were conducted to compare the association and predictive capacity of different obesity indicators for the MONW phenotype. All analyses were stratified by sex. RESULTS: Modified poisson regression analyses revealed that weight, WC, HC, BMI, WHR, WHtR, ABSI, BRI, AVI, WWI, BAI, CI, and WHHR were independently associated with higher risk of the MONW phenotype, regardless of whether they were treated as a continuous or categorical variable (P < 0.05). Notably, BRI demonstrated the strongest association in both men (highest quartile VS lowest quartile; PR = 3.14, 95%CI, 2.49, 3.96; P < 0.001) and women (PR = 4.63, 95%CI, 3.81, 5.62; P < 0.001). Receiver operating characteristic analysis indicated that AUC for the different anthropometric indices ranged from 0.50 to 0.80. BRI and WHtR had the largest AUC in both males (both AUC = 0.733; 95% CI, 0.717, 0.750) and females (both AUC = 0.773; 95% CI, 0.761, 0.786). The optimal cut-off points for BRI, determined by maximizing the Youden's index, were 3.102 (sensitivity: 63.2%, specificity: 36.2%) in males and 3.136 (sensitivity: 68.9%, specificity: 44.2%) in females. Moreover, BRI and WHtR exhibited the highest diagnostic accuracy in younger age groups, specifically those aged 18-34 in both sexes. CONCLUSIONS: BRI emerged as the optimal predictor and independent determinant of the MONW phenotype, regardless of gender. This association was particularly pronounced in young individuals.

Is fluorometric sentinel lymph node biopsy in endometrial cancer necessary?

Objective: In this study, we collected perioperative and postoperative follow-up data from patients with endometrial cancer (EC) at different stages to evaluate the role of sentinel lymph node biopsy (SLNB) in endometrial cancer surgery. Methods: A total of 186 endometrial cancer patients undergoing radical hysterectomy from January 2018 to April 2022 were retrospectively analyzed. Patients were classified into four groups. Group A comprised patients diagnosed with stage IA grade 1 and 2 endometrioid EC who underwent SLNB. Group B comprised patients with stage IA grade 1 and 2 endometrioid EC who did not undergo SLNB. Group C comprised patients with higher-grade endometrioid EC, wherein systematic lymph node dissection was performed based on SLNB results. Group D comprised patients with higher-grade endometrioid EC who did not undergo SLNB and instead underwent direct systematic lymph node dissection. Clinical, pathological data, and follow-up information for all patients were collected. Results: In Group A and B, SLNB was performed on 36 out of 67 patients with IA stage 1 and 2 endometrial cancer, yielding a SLN positivity rate of 5.6%. There were no significant differences observed between the two groups regarding perioperative outcomes and postoperative follow-up. Conversely, among 119 patients with higher-grade endometrial cancer, 52 underwent SLNB, with 20 patients exhibiting SLN positivity, resulting in a SLN positivity rate of 38.4%. However, the decision to undergo SLNB did not yield significant differences in perioperative outcomes and postoperative follow-up among these patients. Conclusion: For stage IA grade 1 and 2 endometrioid EC, the incidence of lymph node positivity is low, omitting SLNB in this subpopulation is a feasible option. In other stages of endometrioid EC, there is no significant difference in perioperative and postoperative follow-up data between patients undergoing routine systematic lymphadenectomy and those undergoing systematic lymphadenectomy based on SLNB results. Therefore, if SLNB is not available, the standard procedure of PLND remains an option to obtain information about lymph node status, despite the surgical complications associated with this procedure.

Prevalence and Risk Factors of Stroke in Inpatients with Type 2 Diabetes Mellitus in China.

OBJECTIVE: The prevalence and the cluster characteristics of risk factors of stroke were assessed in a Chinese diabetic population. METHODS: Clinical data of 30 693 inpatients who were diagnosed with type 2 diabetes mellitus (T2DM) and admitted between 2013 and 2018 were retrospectively analyzed. The age-standardized prevalence of stroke was estimated using the 2010 Chinese population census data, and risk factors were analyzed by multiple imputation and regression. RESULTS: The crude and standardized prevalence rates of stroke in patients with T2DM were 34.4% and 21.5%, respectively, and 85.2% of the stroke patients had ischemic stroke. Nearly half of the patients who experienced stroke had clusters of more than 4 risk factors. Compared with no-risk-factor clustering, the risk of stroke significantly increased 3-4 times in the presence of more than 4 risk-factor clusters (P<0.001). Hypertension was the most common major risk factor for ischemic stroke [odds ratio (OR), 2.34; 95% confidence interval (CI), 2.18-2.50] and hemorrhagic stroke (OR, 3.68; 95% CI 2.95-4.59; P<0.001). Moreover, a 1-standard-deviation increase in fasting blood glucose (FBG) was significantly negatively correlated with ischemic stroke risk, and the same change in FBG was significantly associated with an 8% increased risk of hemorrhagic stroke. CONCLUSION: The prevalence of stroke in patients with T2DM is rather high, and the clustering of risk factors is associated with the development of stroke in T2DM patients. Risk factors differ in different stroke subtypes. Identifying risk factors for a specific high-risk group is necessary.

Machine learning predicts the growth of cyanobacterial genera in river systems and reveals their different environmental responses.

Cyanobacterial blooms are a common and serious problem in global freshwater environments. However, the response mechanisms of various cyanobacterial genera to multiple nutrients and pollutants, as well as the factors driving their competitive dominance, remain unclear or controversial. The relative abundance and cell density of two dominant cyanobacterial genera (i.e., Cyanobium and Microcystis) in river ecosystems along a gradient of anthropogenic disturbance were predicted by random forest with post-interpretability based on physicochemical indices. Results showed that the optimized predictions all reached strong fitting with R2 > 0.75, and conventional water quality indices played a dominant role. One-dimensional and two-dimensional partial dependence plot (PDP) revealed that the responses of Cyanobium and Microcystis to nutrients and temperature were similar, but they showed differences in preferrable nutrient utilization and response to pollutants. Further prediction and PDP for the ratio of Cyanobium and Microcystis unveiled that their distinct responses to PAHs and SPAHs were crucial drivers for their competitive dominance over each other. This study presents a new way for analyzing the response of cyanobacterial genera to multiple environmental factors and their dominance relationships by interpretable machine learning, which is suitable for the identification and interpretation of high-dimensional nonlinear ecosystems with complex interactions.

Association of Body Mass Index and Central Obesity with Spinopelvic Alignment Parameters in a Chinese Population: A Prospective Study.

OBJECTIVE: The purpose of this study was to explore the impact of central obesity on spinal sagittal balance in adults aged 18 and older by examining correlations between waist circumference (WC) and abdominal circumference (AC) and spinopelvic alignment parameters. METHODS: This prospective cohort study included 350 adults aged 18 and older. Participants underwent whole-body biplanar radiography using the EOS imaging system. Spinal and pelvic parameters were measured and correlated with body mass index, WC, and AC. Statistical analyses included one-way analysis of variance, Wilcoxon rank-sum tests for data with nonhomogeneous variances, and chi-squared tests for categorical data. Intra-rater and inter-rater reliability were assessed using intraclass correlation coefficients, with subsequent analyses to explore correlations between body measurements and spinal parameters. RESULTS: The study found significant correlations between increased WC and AC and changes in spinopelvic parameters. However, obesity did not uniformly influence all sagittal alignment parameters. Significant variations in spinal measurements indicate that central obesity plays a role in altering spinal stability and alignment. CONCLUSIONS: The findings highlight the impact of central obesity on spinal alignment and emphasize the importance of considering central obesity in clinical assessments of spinal pathologies. Further research is essential to better understand the relationship between obesity, spinal sagittal balance, and related health conditions.

Plasma proteomics analysis reveals potential biomarkers for intracranial aneurysm formation and rupture.

The aim of this study was to investigate the plasma proteome in individuals with intracranial aneurysms (IAs) and identify biomarkers associated with the formation and rupture of IAs. Proteomic profiles (N = 1069 proteins) were assayed in plasma (N = 120) collected from patients with ruptured and unruptured intracranial aneurysms (RIA and UIA), traumatic subarachnoid hemorrhage (tSAH), and healthy controls (HC) using tandem mass tag (TMT) labeling quantitative proteomics analysis. Gene ontology (GO) and pathway analysis revealed that these relevant proteins were involved in immune response and extracellular matrix organization pathways. Seven candidate biomarkers were verified by ELISA in a completely separate cohort for validation (N = 90). Among them, FN1, PON1, and SERPINA1 can be utilized as diagnosis biomarkers of IA, with a combined area under the ROC curve of 0.891. The sensitivity was 93.33%, specificity was 75.86%, and accuracy was 87.64%. PFN1, ApoA-1, and SERPINA1 can serve as independent risk factors for predicting aneurysm rupture. The combined prediction of aneurysm rupture yielded an area under the ROC curve of 0.954 with a sensitivity of 96.15%, specificity of 81.48%, and accuracy of 88.68%. This prediction model was more effective than PHASES score. In conclusion, high-throughput proteomics analysis with population validation was performed to assess blood-based protein expression characteristics. This revealed the potential mechanism of IA formation and rupture, facilitating the discovery of biomarkers. SIGNIFICANCE: Although the annual rupture rate of small unruptured aneurysms is believed to be minimal, studies have indicated that ruptured aneurysms typically have an average size of 6.28 mm, with 71.8% of them being <7 mm in diameter. Hence, evaluating the possibility of rupture in UIA and making a choice between aggressive treatment and conservative observation emerges as a significant challenge in the management of UIA. No biomarker or scoring system has been able to satisfactorily address this issue to date. It would be significant to develop biomarkers that could be used for early diagnosis of IA as well as for prediction of IA rupture. After TMT proteomics analysis and ELISA validation in independent populations, we found that FN1, PON1, and SERPINA1 can be utilized as diagnostic biomarkers for IA, and PFN1, ApoA-1, and SERPINA1 can serve as independent risk factors for predicting aneurysm rupture. Especially, when combined with ApoA-1, SERPINA1, and PFN1 for predicting IA rupture, the area under the curve (AUC) was 0.954 with a sensitivity of 96.15%, specificity of 81.48%, and accuracy of 88.68%. This prediction model was more effective than PHASES score.

Lower micromolar activity of the antifungal imidazoles on the bacterial-type bifunctional aldehyde/alcohol dehydrogenase (AdhE) in Cryptosporidium parvum and in vitro efficacy against the zoonotic parasite.

Cryptosporidium parvum is a waterborne and foodborne zoonotic protozoan parasite, a causative agent of moderate to severe diarrheal diseases in humans and animals. However, fully effective treatments are unavailable for medical and veterinary uses. There is a need to explore new drug targets for potential development of new therapeutics. Because C. parvum relies on anaerobic metabolism to produce ATP, fermentative enzymes in this parasite are attractive targets for exploration. In this study, we investigated the ethanol-fermentation in the parasite and characterized the basic biochemical features of a bacterial-type bifunctional aldehyde/alcohol dehydrogenase, namely CpAdhE. We also screened 3892 chemical entries from three libraries and identified 14 compounds showing >50% inhibition on the enzyme activity of CpAdhE. Intriguingly, antifungal imidazoles and unsaturated fatty acids are the two major chemical groups among the top hits. We further characterized the inhibitory kinetics of selected imidazoles and unsaturated fatty acids on CpAdhE. These compounds displayed lower micromolar activities on CpAdhE (i.e., IC50 values ranging from 0.88 to 11.02 µM for imidazoles and 8.93 to 35.33 µM for unsaturated fatty acids). Finally, we evaluated the in vitro anti-cryptosporidial efficacies and cytotoxicity of three imidazoles (i.e., tioconazole, miconazole and isoconazole). The three antifungal imidazoles exhibited lower micromolar efficacies against the growth of C. parvum in vitro (EC50 values ranging from 4.85 to 10.41 µM and selectivity indices ranging from 5.19 to 10.95). The results provide a proof-of-concept data to support that imidazoles are worth being further investigated for potential development of anti-cryptosporidial therapeutics.

Novel Bifunctional Conjugates Targeting PD-L1/PARP7 as Dual Immunotherapy for Potential Cancer Treatment.

Bifunctional conjugates targeting PD-L1/PARP7 were designed, synthesized, and evaluated for the first time. Compounds B3 and C6 showed potent activity against PD-1/PD-L1 interaction (IC50 = 0.426 and 0.342 µM, respectively) and PARP7 (IC50 = 2.50 and 7.05 nM, respectively). They also displayed excellent binding affinity with hPD-L1, approximately 100-200-fold better than that of hPD-1. Both compounds restored T-cell function, leading to the increase of IFN-γ secretion. In the coculture assay, B3 and C6 enhanced the killing activity of MDA-MB-231 cells by Jurkat T cells in a concentration-dependent manner. Furthermore, B3 and C6 displayed significant in vivo antitumor efficacy in a melanoma B16-F10 tumor mouse model, more than 5.3-fold better than BMS-1 (a PD-L1 inhibitor) and RBN-2397 (a PARP7i clinical candidate) at the dose of 25 mg/kg, without observable side effects. These results provide valuable insight and understanding for developing bifunctional conjugates for potential anticancer therapy.

PARP-1 dependent cell death pathway (Parthanatos) mediates early brain injury after subarachnoid hemorrhage.

Subarachnoid hemorrhage (SAH) is a neurological condition with high mortality and poor prognosis, and there are currently no effective therapeutic drugs available. Poly (ADP-ribose) polymerase 1 (PARP-1) dependent cell death pathway-parthanatos is closely associated with stroke. We investigated improvements in neurological function, oxidative stress, blood-brain barrier and parthanatos-related protein expression in rats with SAH after intraperitoneal administration of PARP-1 inhibitor (AG14361). Our study found that the expression of parthanatos-related proteins was significantly increased after SAH. Immunofluorescence staining showed increased expression of apoptosis-inducing factor (AIF) in the nucleus after SAH. Administration of PARP-1 inhibitor significantly reduced malondialdehyde (MDA) level and the expression of parthanatos-related proteins. Immunofluorescence staining showed that PARP-1 inhibitor reduced the expression of 8-hydroxy-2' -deoxyguanosine (8-OHdG) and thus reduced oxidative stress. Moreover, PARP-1 inhibitor could inhibit inflammation-associated proteins level and neuronal apoptosis, protect the blood-brain barrier and significantly improve neurological function after SAH. These results suggest that PARP-1 inhibitor can significantly improve SAH, and the underlying mechanism may be through inhibiting parthanatos pathway.