Enhanced anaerobic digestion of waste-activated sludge by thermal-alkali pretreatment: a pilot-scale study.

The composition of waste-activated sludge (WAS) is complex, containing a large amount of harmful substances, which pose a threat to the environment and human health. The reduction and resource utilization of sludge has become a development demand in sludge treatment and disposal. Based on the technical bottlenecks in the practical application of direct anaerobic digestion technology, this study adopted two different thermal and thermal-alkali hydrolysis technologies to pretreat sludge. A pilot-scale experiment was conducted to investigate the experimental conditions, parameters, and effects of two hydrolysis technologies. This study showed that the optimal hydrolysis temperature was 70 °C, the hydrolysis effect and pH can reach equilibrium with the hydrolysis retention time was 4-8 h, and the optimal alkali concentration range was 0.0125-0.015 kg NaOH/kg dry-sludge. Thermal-alkali combination treatment greatly improved the performance of methane production, the addition of NaOH increased methane yield by 31.2% than that of 70 °C thermal hydrolysis. The average energy consumption is 75 kWh/m3 80% water-content sludge during the experiment. This study provides a better pretreatment strategy for exploring efficient anaerobic digestion treatment technologies suitable for southern characteristic sewage sludge.

Three new species of Neohelicomyces (Tubeufiales, Tubeufiaceae) from freshwater and terrestrial habitats in China.

Neohelicomyces species are a group of helicosporous hyphomycetes with the potential to produce secondary metabolites. During our investigation of helicosporous fungi, six collections were isolated from both terrestrial and freshwater habitats in Guizhou Province, China. Based on multigene phylogenetic analysis (ITS, LSU, tef1α and rpb2), coupled with morphological data, three new Neohelicomyces species, viz. N.guizhouensis, N.helicosporus and N.hydei were established. A list of accepted Neohelicomyces species with molecular data was provided. The strain of Neohelicomycespallidus (UAMH 10535) was synonymised under N.denticulatus based on molecular data.

Relating the carbon sources to denitrifying community in full-scale wastewater treatment plants.

Carbon source is a key factor determining the denitrifying effectiveness and efficiency in wastewater treatment plants (WWTPs). Whereas, the relationships between diverse and distinct denitrifying communities and their favorable carbon sources in full-scale WWTPs were not well-understood. This study performed a systematic analysis of the relationships between the denitrifying community and carbon sources by using 15 organic compounds from four categories and activated sludge from 8 full-scale WWTPs. Results showed that, diverse denitrifying bacteria were detected with distinct relative abundances in 8 WWTPs, such as Haliangium (1.98-4.08%), Dechloromonas (2.00-3.01%), Thauera (0.16-1.06%), Zoogloea (0.09-0.43%), and Rhodoferax (0.002-0.104%). Overall, acetate resulted in the highest denitrifying activities (1.21-4.62 mg/L/h/gMLSS), followed by other organic acids (propionate, butyrate and lactate, etc.). Detectable dissimilatory nitrate reduction to ammonium (DNRA) was observed for all 15 carbon sources. Methanol and glycerol resulted in the highest DRNA. Acetate, butyrate, and lactate resulted in the lowest DNRA. Redundancy analysis and 16S cDNA amplicon sequencing suggested that carbon sources within the same category tended to correlate to similar denitrifiers. Methanol and ethanol were primarily correlated to Haliangium. Glycerol and amino acids (glutamate and aspartate) were correlated to Inhella and Sphaerotilus. Acetate, propionate, and butyrate were positively correlated to a wide range of denitrifiers, explaining the high efficiency of these carbon sources. Additionally, even within the same genus, different amplicon sequence variants (ASVs) performed distinctly in terms of carbon source preference and denitrifying capabilities. These findings are expected to benefit carbon source formulation and selection in WWTPs.

Effects of carbonization temperature and time on the characteristics of carbonized sludge.

To investigate the influence of carbonization process parameters on the characteristics of municipal sludge carbonization products, this study selected carbonization temperatures of 300-700 °C and carbonization times of 0.5-1.5 h to carbonize municipal sludge. The results showed that with an increase in temperature and carbonization time, the sludge was carbonized more completely, and the structure and performance characteristics of the sludge changed significantly. Organic matter was continuously cracked, the amorphous nature of the material was reduced, its morphology was transformed into an increasing number of regular crystalline structures, and the content of carbon continued to decrease, from the initial 52.85 to 38.77%, while the content of inorganic species consisting continued to increase. The conductivity was reduced by 87.8%, and the degree of conversion of salt ions into their residual and insoluble states was significant. Natural water absorption in the sludge decreased from 8.13 to 1.29%, and hydrophobicity increased. The dry-basis higher calorific value decreased from 8,703 to 3,574 kJ/kg. Heavy metals were concentrated by a factor of 2-3, but the content of the available state was very low. The results of this study provide important technological support for the selection of suitable carbonization process conditions and for resource utilization.

Enhancing antitumor efficacy of oncolytic virus M1 via albendazole-sustained CD8+ T cell activation.

The immune response plays a crucial role in the functionality of oncolytic viruses. In this study, Albendazole, an antihelminthic drug known to modulate the immune checkpoint PD-L1, was combined with the oncolytic virus M1 (OVM1) to treat mice with either prostate cancer (RM-1) or glioma (GL261) tumors. This combination therapy enhanced anti-tumor effects in immunocompetent mice, but not in immunodeficient ones, without increasing OVM1 replication. Instead, it led to an increase in the number of CD8+ T cells within the tumor, downregulated the expression of PD1 on CD8+ T cells, and upregulated activation markers such as Ki67, CD44, and CD69 and the secretion of cytotoxic factors including interferon (IFN)-γ, granzyme B, and tumor necrosis factor (TNF)-α. Consistently, it enhanced the in vitro tumor-killing activity of lymphocytes from tumor-draining lymph nodes or spleens. The synergistic effect of Albendazole on OVM1 was abolished by depleting CD8+ T cells, suggesting a CD8+ T cell-dependent mechanism. In addition, Albendazole and OVM1 therapy increased CTLA4 expression in the spleen, and the addition of CTLA4 antibodies further enhanced the anti-tumor efficacy in vivo. In summary, Albendazole can act synergistically with oncolytic viruses via CD8+ T cell activation, and the Albendazole/OVM1 combination can overcome resistance to CTLA4-based immune checkpoint blockade therapy.

Identification of two new species and a new host record of Distoseptispora (Distoseptisporaceae, Distoseptisporales, Sordariomycetes) from terrestrial and freshwater habitats in Southern China.

During our investigation of saprophytic fungi in Guizhou and Hainan provinces, China, three hyphomycetes were collected from terrestrial and freshwater habitats. Based on morphological characteristics and phylogenetic analyses of combined ITS, LSU, tef1-α, and rpb2 sequence data, two new species are introduced: Distoseptisporahainanensis and D.lanceolatispora. Additionally, one known species, D.tectonae, previously unreported from Edgeworthiachrysantha, is newly reported. Detailed descriptions, illustrations, and a phylogenetic tree to show the two new species and the new host record of Distoseptispora are provided. In addition, a checklist of Distoseptispora species with their locations, lifestyles, habitats, and hosts is provided.

Overexpression of PtoMYB99 diminishes poplar tolerance to osmotic stress by suppressing ABA and JA biosynthesis.

Drought poses a serious challenge to sustained plant growth and crop yields in the context of global climate change. Drought tolerance in poplars and their underlying mechanisms still remain largely unknown. In this article, we investigated the overexpression of PtoMYB99 - both a drought and abscisic acid (ABA) induced gene constraining drought tolerance in poplars (as compared with wild type poplars). First, we found that PtoMYB99-OE lines exhibited increased stomatal opening and conductance, higher transpiration and photosynthetic rates, as well as reduced levels of ABA and jasmonic acid (JA). Second, PtoMYB99-OE lines accumulated more reactive oxygen species (ROS), including H2O2 and O2-, as well as malonaldehyde (MDA), proline, and soluble sugar under osmotic stress; conversely, the activity of antioxidant enzymes (SOD, POD, and CAT), was weakened in the PtoMYB99-OE lines. Third, the expression of ABA biosynthetic genes, PtoNCED3.1 and PtoNCED3.2, as well as JA biosynthetic genes, PtoOPR3.1 and PtoOPR3.2, was significantly reduced in the PtoMYB99-OE lines under both normal conditions and osmotic stress. Based on our results, we conclude that the overexpression of PtoMYB99 compromises tolerance to osmotic stress in poplar. These findings contribute to the understanding of the role of the MYB genes in drought stress and the biosynthesis of ABA and JA.

Rare loss-of-function variants in matrisome genes are enriched in Ebstein's anomaly.

Ebstein's anomaly, a rare congenital heart disease, is distinguished by the failure of embryological delamination of the tricuspid valve leaflets from the underlying primitive right ventricle myocardium. Gaining insight into the genetic basis of Ebstein's anomaly allows a more precise definition of its pathogenesis. In this study, two distinct cohorts from the Chinese Han population were included: a case-control cohort consisting of 82 unrelated cases and 125 controls without cardiac phenotypes and a trio cohort comprising 36 parent-offspring trios. Whole-exome sequencing data from all 315 participants were utilized to identify qualifying variants, encompassing rare (minor allele frequency < 0.1% from East Asians in the gnomAD database) functional variants and high-confidence (HC) loss-of-function (LoF) variants. Various statistical models, including burden tests and variance-component models, were employed to identify rare variants, genes, and biological pathways associated with Ebstein's anomaly. Significant associations were noted between Ebstein's anomaly and rare HC LoF variants found in genes related to the matrisome, a collection of extracellular matrix (ECM) components. Specifically, 47 genes with HC LoF variants were exclusively or predominantly identified in cases, while nine genes showed such variants in the probands. Over half of unrelated cases (n = 42) and approximately one-third of probands (n = 12) were found to carry one or two LoF variants in these prioritized genes. These results highlight the role of the matrisome in the pathogenesis of Ebstein's anomaly, contributing to a better understanding of the genetic architecture underlying this condition. Our findings hold the potential to impact the genetic diagnosis and treatment approaches for Ebstein's anomaly.

The evolution of artificial oasis over two millennia in Hexi corridor, China.

Artificial oasis is an important geographical unit that is formed by the combination of natural disturbances and human interventions in a fragile ecosystem of arid regions. Therefore, there are important theoretical and practical significance to estimate long term artificial oasis changes and its driving mechanism. We took Hexi Corridor in northwest of China as a case study, based on multi-types of data included documentary reconstruction records, historical archives data and published articles to quantitative analyze the relationship between artificial oasis evolution and a set of natural and human factors over the past two millennia (0-1990 AD). Results showed that artificial oases had a tortuous evolutionary history, experienced three periods of rapid development in the Hexi Corridor. The structural equation model revealed that two human related factors (population density and grain yield) have a significantly greater influence on artificial oasis development than natural variables (temperature). According to current conditions, paradigm shift in human behavior are the prerequisite for oasis evolution and its transition. Furthermore, assessment the scale of artificial oasis based on the human-land relationship is a future direction for research and practice, for allocating water and soil resources rationally, improving the quality of oasis, and controlling the scale of oasis.

Morphophylogenetic evidence reveals four new fungal species within Tetraplosphaeriaceae (Pleosporales, Ascomycota) from tropical and subtropical forest in China.

Tetraplosphaeriaceae (Pleosporales, Ascomycota) is a family with many saprobes recorded from various hosts, especially bamboo and grasses. During a taxonomic investigation of microfungi in tropical and subtropical forest regions of Guizhou, Hainan and Yunnan provinces, China, several plant samples were collected and examined for fungi. Four newly discovered species are described based on morphology and evolutionary relationships with their allies inferred from phylogenetic analyses derived from a combined dataset of LSU, ITS, SSU, and tub2 DNA sequence data. Detailed illustrations, descriptions and taxonomic notes are provided for each species. The four new species of Tetraplosphaeriaceae reported herein are Polyplosphaeriaguizhouensis, Polyplosphaeriahainanensis, Pseudotetraploayunnanensis, and Tetraploahainanensis. A checklist of Tetraplosphaeriaceae species with available details on their ecology is also provided.

Strategy for a Rational Design of Deep-Ultraviolet Nonlinear Optical Materials from Zeolites.

Deep-ultraviolet (DUV) nonlinear optical (NLO) materials play a crucial role in cutting-edge laser technology. In order to solve the serious layered growth tendency of the sole commercial DUV NLO crystal KBe2BO3F2 (KBBF), developing alternative systems of compounds with bulk crystal habits has become an urgent task for practical applications. Herein, a novel strategy was developed by applying non-centrosymmetric (NCS) cancrinite (CAN)-type zincophosphates {Na6(OH)2(H2O)2}Cs2[ZnPO4]6 with bulk-crystal habits as the prototype to design new DUV NLO crystals. Two new anhydrous alkali zincophosphates, namely, {(Li6 -xNaxO)A2}[(ZnPO4)6] (A = Cs, Rb; x = 2-3) crystallizing in the NCS hexagonal space group P63 (no. 173) with a CAN-type framework, were successfully synthesized via a modified fluoro-solvo-hydrothermal method by applying triethylamine (TEA) and concentrated NaF solution as a co-solvent. Interestingly, the rigidity of the NCS CAN-type framework acting as the host ensures the non-centrosymmetry of the resulting new compounds. Meanwhile, the replacement of water molecules by guest cationic species in the channels or cages can greatly improve the thermal stability of the resultant crystal and tune its NLO properties. The synergetic effect of the host framework and the guest species makes the two compounds transparent down to the DUV region (<200 nm) and exhibit SHG effects. Therefore, the proposed rational design strategy of applying the known zeotype NCS frameworks as prototypes together with the modified fluoro-solvo-hydrothermal method opens a great avenue for highly effectively exploring new DUV NLO materials.

Heterozygous nonsense variants in laminin subunit 3α resulting in Ebstein's anomaly.

Ebstein's anomaly is a rare congenital heart disease characterized by tricuspid valve downward displacement and is associated with additional cardiac phenotypes such as left ventricle non-compaction. The genetic basis of Ebstein's anomaly has yet to be fully elucidated, although several genes (e.g., NKX2-5, MYH7, TPM1, and FLNA) may contribute to Ebstein's anomaly. Here, in two Ebstein's anomaly families (a three-generation family and a trio), we identified independent heterozygous nonsense variants in laminin subunit 3 α (LAMA3), cosegregated with phenotypes in families with reduced penetrance. Furthermore, knocking out Lama3 in mice revealed that haploinsufficiency of Lama3 led to Ebstein's malformation of the tricuspid valve and an abnormal basement membrane structure. In conclusion, we identified a novel gene-disease association of LAMA3 implicated in Ebstein's anomaly, and the findings extended our understanding of the role of the extracellular matrix in Ebstein's anomaly etiology.

Two new fungal genera (Diaporthales) found on Dipterocarpaceae in Thailand.

Diaporthales is a species-rich order of fungi that includes endophytes, saprobes, and pathogens associated with forest plants and crops. They may also occur as parasites or secondary invaders of plant tissues injured or infected by other organisms or inhabit living animal and human tissues, as well as soil. Meanwhile, some severe pathogens wipe out large-scale cultivations of profitable crops, timber monocultures, and forests. Based on morphological and phylogenetic analyses of combined ITS, LSU, tef1-α, and rpb2 sequence data, generated using maximum likelihood (ML), maximum parsimony (MP), and MrBayes (BI), we introduce two new genera of Diaporthales found in Dipterocarpaceae in Thailand, namely Pulvinaticonidioma and Subellipsoidispora. Pulvinaticonidioma is characterized by solitary, subglobose, pycnidial, unilocular conidiomata with the internal layers convex and pulvinate at the base; hyaline, unbranched, septate conidiophores; hyaline, phialidic, cylindrical to ampulliform, determinate conidiogenous cells and hyaline, cylindrical, straight, unicellular, and aseptate conidia with obtuse ends. Subellipsoidispora has clavate to broadly fusoid, short pedicellate asci with an indistinct J- apical ring; biturbinate to subellipsoidal, hyaline to pale brown, smooth, guttulate ascospores that are 1-septate and slightly constricted at the septa. Detailed morphological and phylogenetic comparisons of these two new genera are provided in this study.

Molecular evolution in different subtypes of multifocal hepatocellular carcinoma.

BACKGROUND: Multifocal hepatocellular carcinoma (MF-HCC) accounts for > 40% of HCCs, exhibiting a poor prognosis than single primary HCCs. Characterizing molecular features including dynamic changes of mutational signature along with clonal evolution, intrahepatic metastatic timing, and genetic footprint in the preneoplastic stage underlying different subtypes of MF-HCC are important for understanding their molecular evolution and developing a precision management strategy. METHODS: We conducted whole-exome sequencing in 74 tumor samples from spatially distinct regions in 35 resected lesions and adjacent noncancerous tissues from 11 patients, 15 histologically confirmed preneoplastic lesions, and six samples from peripheral blood mononuclear cells. A previously published MF-HCC cohort (n = 9) was included as an independent validation dataset. We combined well-established approaches to investigate tumor heterogeneity, intrahepatic metastatic timing, and molecular footprints in different subtypes of MF-HCCs. RESULTS: We classified MF-HCCs patients into three subtypes, including intrahepatic metastasis, multicentric occurrence, and mixed intrahepatic metastasis and multicentric occurrence. The dynamic changes in mutational signatures between tumor subclonal expansions demonstrated varied etiologies (e.g., aristolochic acid exposure) underlying the clonal progression in different MF-HCC subtypes. Furthermore, the clonal evolution in intrahepatic metastasis exhibited an early metastatic seeding at 10-4-0.01 cm3 in primary tumor volume (below the limits of clinical detection), further validated in an independent cohort. In addition, mutational footprints in the preneoplastic lesions for multicentric occurrence patients revealed common preneoplastic arising clones, evidently being ancestors of different tumor lesions. CONCLUSION: Our study comprehensively characterized the varied tumor clonal evolutionary history underlying different subtypes of MF-HCC and provided important implications for optimizing personalized clinical management for MF-HCC.

The extraction and identification of active components of the sex pheromones of Asian citrus psyllid, Diaphorina citri.

In this study, we extracted and identified the active components of the Asian citrus psyllid, Diaphorina citri sex pheromones to provide a basis for further development of sex attractants. Under laboratory conditions, mating activity in D. citri started 3 d after emergence, which peaked at 6-7 d, and mating activity had no obvious peak during the observed period 7:00-21:00 h. Additionally, D. citri males were attracted to the emanations from conspecific females, especially to the n-hexane extracts of the pheromone. A total of 17 compounds were identified from the n-hexane extracts of female and male D. citri by gas chromatography-mass spectrometer (GC-MS). Among them, 13 compounds were identified from the female D. citri n-hexane extracts, of which 7 (dichloromethane, acetic acid, toluene, butyl acetate, ethyl carbamoylacetate, α-pinene, and 1-nonanal) were not found in the male D. citri n-hexane extracts. In addition, a total of 33 compounds were identified from the solid phase microextraction (SPME) volatiles of the male and female D. citri adults. Among these, 17 compounds were identified from the female D. citri volatiles, of which 6 (cycloheptatriene, 5-methyl-2-phenylindole, 1-dodecanol, cis-11-hexadecena, dodecyl aldehyde, and nerylacetone) were not identified in the volatiles of the D. citri males. It was found that males were significantly attracted to 0.1-10 µL/mL acetic acid and 1-nonanal with the selection rates ranging from 62.04%-70.56% and 62.22%-67.22%, respectively. Therefore, the results of this study suggest that acetic acid and 1-nonanal might be the active compounds of the female D. citri sex pheromones.

High-Sensitivity Wearable Sensor Based On a MXene Nanochannel Self-Adhesive Hydrogel.

To address the shortcomings of traditional filler-based wearable hydrogels, a new type of nanochannel hydrogel sensor is fabricated in this work through a combination of the unique structure of electrospun fiber textile and the properties of a double network hydrogel. Unlike the traditional Ti3C2Tx MXene-based hydrogels, the continuously distributed Ti3C2Tx MXene in the nanochannels of the hydrogel forms a tightly interconnected structure similar to the neuron network. As a result, they have more free space to flip and perform micromovements, which allows one to significantly increase the electrical conductivity and sensitivity of the hydrogel. According to the findings, the Ti3C2Tx MXene nanochannel hydrogel has excellent mechanical properties as well as self-adhesion and antifreezing characteristics. The hydrogel sensor successfully detects different human motions and physiological signals (e.g., low pulse signals) with high stability and sensitivity. Therefore, the proposed Ti3C2Tx MXene-based hydrogel with a unique structure and properties is very promising in the field of flexible wearable devices.

[SigE inhibits DNA damage and participates in the regulation of DNA damage repair in Mycobacterium smegmatis].

Objective To investigate the anti-DNA damage role of Sigma factor E (SigE) and its regulation mechanism of DNA damage repair in Mycobacterium smegmatis(MS). Methods The SigE gene of Mycobacterium smegmatis was cloned into plasmid pMV261 to construct recombinant plasmid pMV261(+)-SigE, and the inserted gene was verified by sequencing. The recombinant plasmid was electrically transformed into Mycobacterium smegmatis to construct SigE over-expression strain, and the expression of SigE was detected by Western blot analysis. The Mycobacterium smegmatis containing pMV261 plasmid was used as the control strain. Growth differences between the two stains were monitored by measuring the 600 nm absorbance (A600) value of the bacterial culture suspension. The survival rate differences between two kinds of strains which were treated with three kinds of DNA damaging agents including ultraviolet ray (UV), cisplatin (DDP), and mitomycin C (MMC) were detected by colony forming unit (CFU) counting assay. DNA damage repair pathways of Mycobacteria were analyzed through bioinformatics and SigE-related genes were screened. The relative expression levels of these genes possibly related to the SigE against DNA damage were detected by real-time fluorescence quantitative PCR. Results SigE over-expression strain pMV261(+)-SigE/MS was constructed and the expression of SigE in Mycobacterium smegmatis was detected. Compared with the control strain, the SigE over-expression strain grew more slowly and entered the growth plateau later; survival rate analysis found that SigE over-expression strain was more resistant to three DNA damaging agents including UV, DDP, and MMC. Bioinformatic analysis indicated that SigE gene was closely related to DNA damage repair genes recA, single-strand DNA binding protein, (ssb), and dnaE2; the expression levels of recA, dnaE2, and ssb in SigE over-expression strain all increased with varying degrees compared with those in the control strain. Conclusion SigE plays an important role in inhibiting the DNA damage of Mycobacterium smegmatis, whose mechanism is closely related to the regulation of DNA damage repair.

Ca-La layered double hydroxide (LDH) for selective and efficient removal of phosphate from wastewater.

Adsorption showed advantages in removing phosphorus (P) at low concentrations. Desirable adsorbents should have sufficiently high adsorption capacity and selectivity. In this study, a Ca-La layered double hydroxide (LDH) was synthesized for the first time by using a simple hydrothermal coprecipitation method for phosphate removal from wastewater. A maximum adsorption capacity of 194.04 mgP/g was achieved, ranking on the top of known LDHs. Adsorption kinetic experiments showed that 0.02 g/L Ca-La LDH could effectively reduce PO43-P from 1.0 to <0.02 mg/L within 30 min. With the copresence of bicarbonate and sulfate at concentrations 17.1 and 35.7 times of that of PO43-P, the Ca-La LDH showed promising selectivity towards phosphate (with a reduction in the adsorption capacity of <13.6%). In addition, four other (Mg-La, Co-La, Ni-La, and Cu-La) LDHs containing different divalent metal ions were synthesized by using the same coprecipitation method. Results showed much higher P adsorption performance of the Ca-La LDH than those LDHs. Field Emission Electron Microscopy (FE-SEM)-Energy Dispersive Spectroscopy (EDS), X-ray Diffraction (XRD), X-ray Photoelectron Spectroscopy (XPS), Fourier Transform Infrared Spectroscopy (FTIR), and mesoporous analysis were performed to characterize and compare the adsorption mechanisms of different LDHs. The high adsorption capacity and selectivity of the Ca-La LDH were mainly explained by selective chemical adsorption, ion exchange, and inner sphere complexation.

A comparison of algorithms for identifying copy number variants in family-based whole-exome sequencing data and its implications in inheritance pattern analysis.

There remain challenges in accurately identifying constitutional or germline copy number variants (gCNVs) based on whole-exome sequencing data that have implications for genetic diagnosis for 'rare undiagnosed disease' in the clinical setting. Although multiple algorithms have been proposed, a systematic comparison of these algorithms for calling gCNVs and analyzing inherited pattern have yet to be fully conducted. Therefore, we empirically compared seven exome-based algorithms, including XHMM, CLAMMS, CODEX2, ExomeDepth, DECoN, CN.MOPS, and GATK gCNV, for calling gCNVs in 151 individuals from 44 pedigrees, together with the gold standard of genotyping-derived gCNVs in the same cohort for the performance assessment. These algorithms demonstrated varied powers in identifying gCNVs, although the distribution of gCNVs size was similar. The number of shared gCNVs across these algorithms was limited (e.g., only four gCNVs shared among seven algorithms); however, several algorithms showed varying degrees of consistency (e.g., 1,843 gCNVs shared between DECoN and ExomeDepth). CLAMMS and CODEX2 outperformed the remaining algorithms according to a relatively higher F-score (i.e., 0.145 and 0.152, respectively). In addition, these algorithms exhibited different Mendelian inconsistencies of gCNVs and significant challenges remained in inheritance pattern analysis. In conclusion, selecting good algorithms may have important implications in gCNVs-based inheritance pattern analysis for family-based studies.

Inhibition of mycobacteria proliferation in macrophages by low cisplatin concentration through phosphorylated p53-related apoptosis pathway.

BACKGROUND: Drug resistance is a prominent problem in the treatment of tuberculosis, so it is urgent to develop new anti- tuberculosis drugs. Here, we investigated the effects and mechanisms of cisplatin (DDP) on intracellular Mycobacterium smegmatis to tap the therapeutic potential of DDP in mycobacterial infection. RESULTS: Macrophages infected with Mycobacterium smegmatis were treated with DDP alone or combined with isoniazid or rifampicin. The results showed that the bacterial count in macrophages decreased significantly after DDP (≤ 6 µg/mL) treatment. When isoniazid or rifampicin was combined with DDP, the number of intracellular mycobacteria was also significantly lower than that of isoniazid or rifampicin alone. Apoptosis of infected cells increased after 24 h of DDP treatment, as shown by flow cytometry and transmission electron microscopy detection. Transcriptome sequencing showed that there were 1161 upregulated and 645 downregulated differentially expressed genes (DEGs) between the control group and DDP treatment group. A Trp53-centered protein interaction network was found based on the top 100 significant DEGs through STRING and Cytoscape software. The expression of phosphorylated p53, Bax, JAK, p38 MAPK and PI3K increased after DDP treatment, as shown by Western blot analysis. Inhibitors of JAK, PI3K or p38 MAPK inhibited the increase in cell apoptosis and the reduction in the intracellular bacterial count induced by DDP. The p53 promoter Kevetrin hydrochloride scavenges intracellular mycobacteria. If combined with DDP, Kevetrin hydrochloride could increase the effect of DDP on the elimination of intracellular mycobacteria. In conclusion, DDP at low concentrations could activate the JAK, p38 MAPK and PI3K pathways in infected macrophages, promote the phosphorylation of p53 protein, and increase the ratio of Bax to Bcl-2, leading to cell apoptosis, thus eliminating intracellular bacteria and reducing the spread of mycobacteria. CONCLUSION: DDP may be a new host-directed therapy for tuberculosis treatment, as well as the p53 promoter Kevetrin hydrochloride.