PARP inhibitors suppress tumours via centrosome error-induced senescence independent of DNA damage response.

BACKGROUND: Poly(ADP-ribose) polymerase (PARP) inhibitors have emerged as promising chemotherapeutic drugs primarily against BRCA1/2-associated tumours, known as synthetic lethality. However, recent clinical trials reported patients' survival benefits from PARP inhibitor treatments, irrelevant to homologous recombination deficiency. Therefore, revealing the therapeutic mechanism of PARP inhibitors beyond DNA damage repair is urgently needed, which can facilitate precision medicine. METHODS: A CRISPR-based knock-in technology was used to establish stable BRCA1 mutant cancer cells. The effects of PARP inhibitors on BRCA1 mutant cancer cells were evaluated by biochemical and cell biological experiments. Finally, we validated its in vivo effects in xenograft and patient-derived xenograft (PDX) tumour mice. FINDINGS: In this study, we uncovered that the majority of clinical BRCA1 mutations in breast cancers were in and near the middle of the gene, rather than in essential regions for DNA damage repair. Representative mutations such as R1085I and E1222Q caused transient extra spindle poles during mitosis in cancer cells. PAR, which is synthesized by PARP2 but not PARP1 at mitotic centrosomes, clustered these transient extra poles, independent of DNA damage response. Common PARP inhibitors could effectively suppress PARP2-synthesized PAR and induce cell senescence by abrogating the correction of mitotic extra-pole error. INTERPRETATION: Our findings uncover an alternative mechanism by which PARP inhibitors efficiently suppress tumours, thereby pointing to a potential new therapeutic strategy for centrosome error-related tumours. FUNDING: Funded by National Natural Science Foundation of China (NSFC) (T2225006, 82272948, 82103106), Beijing Municipal Natural Science Foundation (Key program Z220011), and the National Clinical Key Specialty Construction Program, P. R. China (2023).

Protocol for identifying metabolite biomarkers in patient uterine fluid for early ovarian cancer detection.

High mortality of ovarian cancer (OC) is primarily attributed to the lack of effective early detection methods. Uterine fluid, pooling molecules from neighboring ovaries, presents an organ-specific advantage over conventional blood samples. Here, we present a protocol for identifying metabolite biomarkers in uterine fluid for early OC detection. We describe steps for uterine fluid collection from patients, metabolite extraction, metabolomics experiments, and candidate metabolite biomarker screening. This standardized workflow holds the potential to achieve early OC diagnosis in clinical practice. For complete details on the use and execution of this protocol, please refer to Wang et al.1.

Profiling the metabolome of uterine fluid for early detection of ovarian cancer.

Ovarian cancer (OC) causes high mortality in women because of ineffective biomarkers for early diagnosis. Here, we perform metabolomics analysis on an initial training set of uterine fluid from 96 gynecological patients. A seven-metabolite-marker panel consisting of vanillylmandelic acid, norepinephrine, phenylalanine, beta-alanine, tyrosine, 12-S-hydroxy-5,8,10-heptadecatrienoic acid, and crithmumdiol is established for detecting early-stage OC. The panel is further validated in an independent sample set from 123 patients, discriminating early OC from controls with an area under the curve (AUC) of 0.957 (95% confidence interval [CI], 0.894-1). Interestingly, we find elevated norepinephrine and decreased vanillylmandelic acid in most OC cells, resulting from excess 4-hydroxyestradiol that antagonizes the catabolism of norepinephrine by catechol-O-methyltransferase. Moreover, exposure to 4-hydroxyestradiol induces cellular DNA damage and genomic instability that could lead to tumorigenesis. Thus, this study not only reveals metabolic features in uterine fluid of gynecological patients but also establishes a noninvasive approach for the early diagnosis of OC.

A 1,2,3-Triazole Derivative of Quinazoline Exhibits Antitumor Activity by Tethering RNF168 to SQSTM1/P62.

Quinazoline and its derivatives have drawn much attention in the development of potential antitumor agents. Here, we synthesized a series of 1,2,3-triazole derivatives of quinazoline at the C6 position and evaluated for their cytotoxic activity in various human cancer cell lines. We found that compound 5a was the most cytotoxic to HCT-116 cells (IC50, 0.36 µM). Target profiling found that 5a directly binds to both the autophagy-associated protein SQSTM1/P62 and the E3 ligase RNF168, promoting their interaction. Consistently, 5a treatment induces a decrease in RNF168-mediated H2A ubiquitination and compromises homologous recombination-mediated DNA repair, thus increasing the sensitivity of HCT-116 to X-ray radiation. Moreover, 5a suppressed xenografted tumor growth in mice in a dose-dependent manner. Taken together, the 1,2,3-triazole derivative of quinazoline 5a may serve as a novel compound for tumor therapy based on its role in promoting a P62/RNF168 interaction.

The Apoptotic Resistance of BRCA1-Deficient Ovarian Cancer Cells is Mediated by cAMP.

Breast cancer type 1 susceptibility protein (BRCA1) is essential for homologous recombination repair of DNA double-strand breaks. Loss of BRCA1 is lethal to embryos due to extreme genomic instability and the activation of p53-dependent apoptosis. However, the apoptosis is resisted in BRCA1-deficient cancer cells even though their p53 is proficient. In this study, by analysis of transcriptome data of ovarian cancer patients bearing BRCA1 defects in TCGA database, we found that cAMP signaling pathway was significantly activated. Experimentally, we found that BRCA1 deficiency caused an increased expression of ADRB1, a transmembrane receptor that can promote the generation of cAMP. The elevated cAMP not only inhibited DNA damage-induced apoptosis through abrogating p53 accumulation, but also suppressed the proliferation of cytotoxic T lymphocytes by enhancing the expression of immunosuppressive factors DKK1. Inhibition of ADRB1 effectively killed cancer cells by abolishing the apoptotic resistance. These findings uncover a novel mechanism of apoptotic resistance in BRCA1-deficient ovarian cancer cells and point to a potentially new strategy for treating BRCA1-mutated tumors.

Cinnamaldehyde prevents intergenerational effect of paternal depression in mice via regulating GR/miR-190b/BDNF pathway.

Paternal stress exposure-induced high corticosterone (CORT) levels may contribute to depression in offspring. Clinical studies disclose the association of depressive symptoms in fathers with their adolescent offspring. However, there is limited information regarding the intervention for intergenerational inheritance of depression. In this study we evaluated the intervention of cinnamaldehyde, a major constituent of Chinese herb cinnamon bark, for intergenerational inheritance of depression in CORT- and CMS-induced mouse models of depression. Depressive-like behaviors were induced in male mice by injection of CORT (20 mg·kg-1·d-1, sc) for 6 weeks or by chronic mild stress (CMS) for 6 weeks. We showed that co-administration of cinnamaldehyde (10, 20, or 40 mg·kg-1·d-1, ig) for 6 weeks in F0 males prevented the depressive-like phenotypes of F1 male offspring. In addition, co-administration of cinnamaldehyde (20 mg·kg-1·d-1, ig) for 4 weeks significantly ameliorated depressive-like behaviors of chronic variable stress (CVS)-stimulated F1 offspring born to CMS mice. Notably, cinnamaldehyde had no reproductive toxicity, while positive drug fluoxetine showed remarkable reproductive toxicity. We revealed that CMS and CORT significantly reduced testis glucocorticoid receptor (GR) expression, and increased testis and sperm miR-190b expression in F0 depressive-like models. Moreover, pre-miR-190b expression was upregulated in testis of F0 males. The amount of GR on miR-190b promoter regions was decreased in testis of CORT-stimulated F0 males. Cinnamaldehyde administration reversed CORT-induced GR reduction in testis, miR-190b upregulation in testis and sperm, pre-miR-190b upregulation in testis, and the amount of GR on miR-190b promoter regions of F0 males. In miR-190b-transfected Neuro 2a (N2a) cells, we demonstrated that miR-190b might directly bind to the 3'-UTR of brain-derived neurotrophic factor (BDNF). In the hippocampus of F1 males of CORT- or CMS-induced depressive-like models, increased miR-190b expression was accompanied by reduced BDNF and GR, which were ameliorated by cinnamaldehyde. In conclusion, cinnamaldehyde is a potential intervening agent for intergenerational inheritance of depression, probably by regulating GR/miR-190b/BDNF pathway.

A synergetic effect of BARD1 mutations on tumorigenesis.

To date, a large number of mutations have been screened from breast and ovarian cancer patients. However, most of them are classified into benign or unidentified alterations due to their undetectable phenotypes. Whether and how they could cause tumors remains unknown, and this significantly limits diagnosis and therapy. Here, in a study of a family with hereditary breast and ovarian cancer, we find that two BARD1 mutations, P24S and R378S, simultaneously exist in cis in surviving cancer patients. Neither of the single mutations causes a functional change, but together they synergetically impair the DNA damage response and lead to tumors in vitro and in vivo. Thus, our report not only demonstrates that BARD1 defects account for tumorigenesis but also uncovers the potential risk of synergetic effects between the large number of cis mutations in individual genes in the human genome.

Pilot study of nitrogen removal from landfill leachate by stable nitritation-denitrification based on zeolite biological aerated filter.

A pilot (about 1 m3/d) process consisting of pre-denitrification and zeolite biological aerated filter (ZBAF) was established and run for nitrogen removal of landfill leachate. The results showed that stable nitritation and denitrification was achieved for landfill leachate with removal efficiency of Chemical Oxygen Demand (CODCr), ammonium and total nitrogen (TN) of 53.2 ±â€¯3.0%, 93.5 ±â€¯2.4% and 74.7 ±â€¯9.4%, respectively. Based on the ammonium adsorption equilibrium by zeolite, stable free ammonia could be maintained for inhibition of nitrite oxidizing bacteria (NOB) and dominance of ammonia oxidizing bacteria (AOB) in ZBAF, resulting in efficient nitritation with a nitrite accumulation ratio higher than 90.0% and an average nitrite production rate of 1.387 kg NO2--N m-3 day-1. High-throughput sequencing analysis further revealed enrichment of AOB and elimination of NOB in ZBAF. Compared to two-stage anoxic-oxic process, the pilot-scale process could save approximate 5000 mg/L glucose (about 3.10 US dollar/m3) with almost similar TN removal performance. All results obtained demonstrated the feasibility of the pilot process, which might be highly promising for the nitritation and denitrification of low C/N landfill leachate in the future.

An in vitro model of foam cell formation induced by a stretchable microfluidic device.

Although a variety of animal models of atherosclerosis have been developed, these models are time-consuming and costly. Here, we describe an in vitro model to induce foam cell formation in the early stage of atherosclerosis. This model is based on a three-dimension co-culture system in a stretchable microfluidic device. An elastic membrane embedded in the microfluidic device is capable of delivering nonuniform strain to vascular smooth muscle cells, endothelial cells and monocytes adhering thereto, which are intended to mimic the biological environment of blood vessels. Under low-density lipoprotein and stretch treatment, foam cell formation was successfully induced in co-culture with changes in mRNA and protein expression of some related key factors. Subsequently, the model was used to assess the inhibitory effect of atorvastatin on foam cell formation. The results obtained indicate that atorvastatin has a significantly dose-dependent inhibition of foam cell formation, which can be explained by the changes in mRNA and protein expression of the related factors. In principle, the model can be used to study the role of different types of cells in the formation of foam cells, as well as the evaluation of anti-atherosclerotic drugs.

Rapid start-up and performance of denitrifying granular sludge in an upflow sludge blanket (USB) reactor treating high concentration nitrite wastewater.

Denitrifying granular sludge reactor holds better nitrogen removal efficiency than other kinds of denitrifying reactors, while this reactor commonly needs seeding anaerobic granular sludge and longer period for start-up in practice, which restricted the application of denitrifying granular sludge reactor. This study presented a rapid and stable start-up method for denitrifying granular sludge. An upflow sludge blanket (USB) reactor with packings was established with flocculent activated sludge for treatment of high concentration nitrite wastewater. Results showed mature denitrifying granular sludge appeared only after 15 days with highest nitrogen removal rate of 5.844 kg N/(m3 day), which was much higher than that of compared anoxic sequencing batch reactor (ASBR). No significant nitrite inhibition occurred in USB and denitrification performance was mainly influenced by hydraulic retention time, influent C/N ratio and internal reflux ratio. Hydraulic shear force created by upflow fluid, shearing of gaseous products and stable microorganisms adhesion on the packings might be the reasons for rapid achievement of granular sludge. Compared to inoculated sludge and ASBR, remarkable microbial communitiy variations were detected in USB. The dominance of Proteobacteria and Bacteroidetes and enrichment of species Pseudomonas_stutzeri should be responsible for the excellent denitrification performance, which further verified the feasibility of start-up method.

Partial nitrification performance and mechanism of zeolite biological aerated filter for ammonium wastewater treatment.

A zeolite biological aerated filter (ZBAF) with continuous feeding was successfully applied for achieving stable partial nitrification. Excellent nitrite accumulation (higher than 98.0%) and high nitrite/nitrate production rate (NPR) (approximately 0.760kg/m3/d) were obtained with increase influent ammonium concentration from 250 to 550mg/L within a nitrogen loading rate (NLR) of 0.854-1.200kg/m3/d. Owning to the adsorption of zeolite to ammonium, free ammonia (FA) concentration could remain at an appropriate range for inhibition of nitrite oxidizing bacteria (NOB) and dominance of ammonia-oxidizing bacteria (AOB), which should be responsible for the excellent partial nitrification realized in ZBAF. Kinetic study showed that the production of nitrite in ZBAF followed the zero-order kinetics model and high-throughput sequencing analysis further presented the enrichment of AOB and inhibition of NOB in ZBAF. All the results demonstrated that ZBAF hold a great potential in the application of partial nitrification for ammonium wastewater treatment.

Regulation of energy homeostasis by the ubiquitin-independent REGγ proteasome.

Maintenance of energy homeostasis is essential for cell survival. Here, we report that the ATP- and ubiquitin-independent REGγ-proteasome system plays a role in maintaining energy homeostasis and cell survival during energy starvation via repressing rDNA transcription, a major intracellular energy-consuming process. Mechanistically, REGγ-proteasome limits cellular rDNA transcription and energy consumption by targeting the rDNA transcription activator SirT7 for ubiquitin-independent degradation under normal conditions. Moreover, energy starvation induces an AMPK-directed SirT7 phosphorylation and subsequent REGγ-dependent SirT7 subcellular redistribution and degradation, thereby further reducing rDNA transcription to save energy to overcome cell death. Energy starvation is a promising strategy for cancer therapy. Our report also shows that REGγ knockdown markedly improves the anti-tumour activity of energy metabolism inhibitors in mice. Our results underscore a control mechanism for an ubiquitin-independent process in maintaining energy homeostasis and cell viability under starvation conditions, suggesting that REGγ-proteasome inhibition has a potential to provide tumour-starving benefits.

Effect of Tourmaline-Doped on the Far Infrared Emission of Iron Ore Tailings Ceramics.

Iron ore tailings as secondary resources have been of great importance to many countries in the world. Their compositions are similar to that of infrared emission ceramics, but there are few reports about it. In addition, tourmaline has high infrared emission properties due to its unique structure. With the purpose of expanding functional utilization of iron ore tailings, as well as reducing the production cost of far infrared ceramics, a new kind of far infrared emission ceramics was prepared by using iron ore tailings, calcium carbonate, silica, and natural tourmaline. The ceramics powders were characterized by Fourier transform infrared spectroscope, X-ray diffraction and scanning electron microscopy, respectively. The results show that after being sintered at 1065 °C, the percentage of pseudobrookite and lattice strain of samples increased with increasing the elbaite content. Furthermore, the added tourmaline was conducive to the densification sintering of ceramics. The appearance of Li-O vibration at 734.73 cm-1, as well as the strengthened Fe-O vibration at 987.68 cm-1 were attributed to the formation of Li0.375Fe1.23Ti1.4O5 solid solution, which led the average far infrared emissivity of ceramics increase from 0.861 to 0.906 within 8-14 µm.

Loss of KLF14 triggers centrosome amplification and tumorigenesis.

Centrosome amplification is frequent in cancer, but the underlying mechanisms remain unclear. Here we report that disruption of the Kruppel-like factor 14 (KLF14) gene in mice causes centrosome amplification, aneuploidy and spontaneous tumorigenesis. Molecularly, KLF14 functions as a transcriptional repressor of Plk4, a polo-like kinase whose overexpression induces centrosome overduplication. Transient knockdown of KLF14 is sufficient to induce Plk4-directed centrosome amplification. Clinically, KLF14 transcription is significantly downregulated, whereas Plk4 transcription is upregulated in multiple types of cancers, and there exists an inverse correlation between KLF14 and Plk4 protein expression in human breast and colon cancers. Moreover, KLF14 depletion promotes AOM/DSS-induced colon tumorigenesis. Our findings reveal that KLF14 reduction serves as a mechanism leading to centrosome amplification and tumorigenesis. On the other hand, forced expression of KLF14 leads to mitotic catastrophe. Collectively, our findings identify KLF14 as a tumour suppressor and highlight its potential as biomarker and therapeutic target for cancer.

Application of integrated ozone biological aerated filters and membrane filtration in water reuse of textile effluents.

A combined process including integrated ozone-BAFs (ozone biological aerated filters) and membrane filtration was first applied for recycling textile effluents in a cotton textile mill with capacity of 5000 m(3)/d. Influent COD (chemical oxygen demand) in the range of 82-120 mg/L, BOD5 (5-day biochemical oxygen demand) of 12.6-23.1 mg/L, suspended solids (SSs) of 38-52 mg/L and color of 32-64° were observed during operation. Outflows with COD≤45 mg/L, BOD5≤7.6 mg/L, SS≤15 mg/L, color≤8° were obtained after being decontaminated by ozone-BAF with ozone dosage of 20-25 mg/L. Besides, the average removal rates of PVA (polyvinyl alcohol) and UV254 were 100% and 73.4% respectively. Permeate water produced by RO (reverse osmosis) could be reused in dyeing and finishing processes, while the RO concentrates could be discharged directly under local regulations with COD≤100 mg/L, BOD5≤21 mg/L, SS≤52 mg/L, color≤32°. Results showed that the combined process could guarantee water reuse with high quality, and solve the problem of RO concentrate disposal.

Combined processes of two-stage Fenton-biological anaerobic filter-biological aerated filter for advanced treatment of landfill leachate.

There are numerous non-biodegradable organic materials in the mature landfill leachate. To meet the new discharge standard of China, additional advanced treatment is needed for the effluent from the biological treatment processes of leachate. In this study, a combined process including two stages of "Fenton-biological anaerobic filter (BANF)-biological aerated filter (BAF)" was evaluated to address the advanced treatment need. The Fenton oxidation was applied to reduce chemical oxygen demand (COD) and enhance biodegradability of refractory organics, and the BANF-BAF process was then applied to remove the total nitrogen (TN). The treatment achieved effluent concentrations of COD<70 mg/L, TN<40 mg/L and NH(3)-N<10 mg/L. The removal efficiency of COD and TN were 96.1% and 95.9%, respectively. The effluent quality met the new discharge standard for Pollution Control on the Landfill Site of Municipal Solid of PR China (GB16889-2008). The operation cost of these processes was about 36.1CHY/t (5.70USD/t).

High viral load of human bocavirus correlates with duration of wheezing in children with severe lower respiratory tract infection.

BACKGROUND: Human bocavirus (HBoV) is a newly discovered parvovirus and increasing evidences are available to support its role as an etiologic agent in lower respiratory tract infection (LRTI). The objective of this study is to assess the impact of HBoV viral load on clinical characteristics in children who were HBoV positive and suffered severe LRTI. METHODS: Lower respiratory tract aspirates from 186 hospitalized children with severe LRTI were obtained by bronchoscopy. HBoVs were detected by real-time PCR and other 10 infectious agents were examined using PCR and/or direct fluorescent assay. RESULTS: Thirty-one patients (24.6%) were tested positive for HBoV in the respiratory tract aspirates. Fifteen samples had a high viral load (>10(4) copies/mL) and the other sixteen samples had a low viral load (<10(4) copies/mL). The duration of presented wheezing and hospitalization was longer in children with high viral load of HBoV than that in children with low viral load. The days of wheezing showed a correlation with viral load of HBoV. CONCLUSION: We confirmed that HBoV was frequently detected in patients with severe LRTI. Wheezing was one of the most common symptoms presented by patients with positive HBoV. A high HBoV viral load could be an etiologic agent for LRTI, which led to more severe lower respiratory tract symptom, longer duration of wheezing and hospitalization.

Pilot study on the advanced treatment of landfill leachate using a combined coagulation, fenton oxidation and biological aerated filter process.

Mature landfill leachate is typically non-biodegradable. A combination process was developed that includes coagulation, Fenton oxidation, and biological aerated filtering to treat biologically-produced effluent. In this process, coagulation and Fenton oxidation were applied in order to reduce chemical oxygen demand (COD) organic load, and enhance biodegradability. Poly-ferric sulfate (PFS) at 600 mgl(-1) was found to be a suitable dosage for coagulation. For Fenton oxidation, an initial pH of 5, a total reaction time of 3h, and an H(2)O(2) dosage of 5.4mmoll(-1), with a (H(2)O(2))/n(Fe(2+)) ratio of 1.2 and two-step dosing were selected to achieve optimal oxidation. Under these optimal coagulation and Fenton oxidation conditions, the COD removal ratios were found to be 66.67% and 56%, respectively. Following pretreatment with coagulation and Fenton oxidation, the landfill leachate was further treated using a biological aerated filter (BAF). Our results show that COD was reduced to 75mgl(-1), and the color was less than 10 degrees.