GGC repeat expansion in NOTCH2NLC induces dysfunction in ribosome biogenesis and translation.

GGC repeat expansion in the 5' untranslated region (UTR) of NOTCH2NLC is associated with a broad spectrum of neurological disorders, especially neuronal intranuclear inclusion disease (NIID). Studies have found that GGC repeat expansion in NOTCH2NLC induces the formation of polyglycine (polyG)-containing protein, which is involved in the formation of neuronal intranuclear inclusions. However, the mechanism of neurotoxicity induced by NOTCH2NLC GGC repeats is unclear. Here, we used NIID patient-specific induced pluripotent stem cell (iPSC)-derived 3D cerebral organoids (3DCOs) and cellular models to investigate the pathophysiological mechanisms of NOTCH2NLC GGC repeat expansion. IPSC-derived 3DCOs and cellular models showed the deposition of polyG-containing intranuclear inclusions. The NOTCH2NLC GGC repeats could induce the upregulation of autophagic flux, enhance integrated stress response and activate EIF2α phosphorylation. Bulk RNA sequencing for iPSC-derived neurons and single-cell RNA sequencing (scRNA-seq) for iPSC-derived 3DCOs revealed that NOTCH2NLC GGC repeats may be associated with dysfunctions in ribosome biogenesis and translation. Moreover, NOTCH2NLC GGC repeats could induce the NPM1 nucleoplasm translocation, increase nucleolar stress, impair ribosome biogenesis and induce ribosomal RNA sequestration, suggesting dysfunction of membraneless organelles in the NIID cellular model. Dysfunctions in ribosome biogenesis and phosphorylated EIF2α and the resulting increase in the formation of G3BP1-positive stress granules may together lead to whole-cell translational inhibition, which may eventually cause cell death. Interestingly, scRNA-seq revealed that NOTCH2NLC GGC repeats may be associated with a significantly decreased proportion of immature neurons while 3DCOs were developing. Together, our results underscore the value of patient-specific iPSC-derived 3DCOs in investigating the mechanisms of polyG diseases, especially those caused by repeats in human-specific genes.

Shared Genetic Architecture between Parkinson's Disease and Brain Structural Phenotypes.

BACKGROUND: Patients with Parkinson's disease (PD) have consistently demonstrated brain structure abnormalities, indicating the presence of shared etiological and pathological processes between PD and brain structures; however, the genetic relationship remains poorly understood. OBJECTIVE: The aim of this study was to investigate the extent of shared genetic architecture between PD and brain structural phenotypes (BSPs) and to identify shared genomic loci. METHODS: We used the summary statistics from genome-wide association studies to conduct MiXeR and conditional/conjunctional false discovery rate analyses to investigate the shared genetic signatures between PD and BSPs. Subsequent expression quantitative trait loci mapping in the human brain and enrichment analyses were also performed. RESULTS: MiXeR analysis identified genetic overlap between PD and various BSPs, including total cortical surface area, average cortical thickness, and specific brain volumetric structures. Further analysis using conditional false discovery rate (FDR) identified 21 novel PD risk loci on associations with BSPs at conditional FDR < 0.01, and the conjunctional FDR analysis demonstrated that PD shared several genomic loci with certain BSPs at conjunctional FDR < 0.05. Among the shared loci, 16 credible mapped genes showed high expression in the brain tissues and were primarily associated with immune function-related biological processes. CONCLUSIONS: We confirmed the polygenic overlap with mixed directions of allelic effects between PD and BSPs and identified multiple shared genomic loci and risk genes, which are likely related to immune-related biological processes. These findings provide insight into the complex genetic architecture associated with PD. © 2023 International Parkinson and Movement Disorder Society.

Urinary exosome proteins PAK6 and EGFR as noninvasive diagnostic biomarkers of diabetic nephropathy.

OBJECTIVE: The actin cytoskeleton plays an essential role in maintaining podocyte functions. However, whether the urinary exosome proteins related to the regulation of the actin cytoskeleton are changed in diabetic nephropathy (DN) is still unknown. This study was to investigate the possibility that related proteins can be applied as diagnostic biomarkers for DN. METHODS: Urinary exosomes were obtained from 144 participants (Discovery phase: n = 72; Validation phase: n = 72) by size exclusion chromatography methods. Proteomic analysis of urinary exosome by LC-MS/MS. Western blot and ELISA were applied to validate the selected urinary exosome proteins. The clinical value of selected urinary exosome proteins was evaluated using correlation and receiver operating characteristic curve analyses. RESULTS: Fifteen urinary proteins related to the regulation of the actin cytoskeleton were identified in urinary exosomes. Three upregulated proteins were selected, including Serine/threonine-protein kinase PAK6 (PAK6), Epidermal growth factor receptor (EGFR), and SHC-transforming protein 1(SHC1). The expression level of PAK6 and EGFR was negatively correlated with estimated glomerular filtration rate and positively correlated with serum creatinine levels. For diagnosing DN in the discovery phase: the area under curve (AUC) of PAK6 was 0.903, EGFR was 0.842, and the combination of two proteins was 0.912. These better performances were also observed in the validation phase (For PAK6: AUC = 0.829; For EGFR: AUC = 0.797; For PAK6 + EGFR: AUC = 0.897). CONCLUSIONS: Urinary exosome proteins PAK6 and EGFR may be promising and noninvasive biomarkers for diagnosing DN.

Identifying causal genes for migraine by integrating the proteome and transcriptome.

BACKGROUND: While previous genome-wide association studies (GWAS) have identified multiple risk variants for migraine, there is a lack of evidence about how these variants contribute to the development of migraine. We employed an integrative pipeline to efficiently transform genetic associations to identify causal genes for migraine. METHODS: We conducted a proteome-wide association study (PWAS) by combining data from the migraine GWAS data with proteomic data from the human brain and plasma to identify proteins that may play a role in the risk of developing migraine. We also combined data from GWAS of migraine with a novel joint-tissue imputation (JTI) prediction model of 17 migraine-related human tissues to conduct transcriptome-wide association studies (TWAS) together with the fine mapping method FOCUS to identify disease-associated genes. RESULTS: We identified 13 genes in the human brain and plasma proteome that modulate migraine risk by regulating protein abundance. In addition, 62 associated genes not reported in previous migraine TWAS studies were identified by our analysis of migraine using TWAS and fine mapping. Five genes including ICA1L, TREX1, STAT6, UFL1, and B3GNT8 showed significant associations with migraine at both the proteome and transcriptome, these genes are mainly expressed in ependymal cells, neurons, and glial cells, and are potential target genes for prevention of neuronal signaling and inflammatory responses in the pathogenesis of migraine. CONCLUSIONS: Our proteomic and transcriptome findings have identified disease-associated genes that may give new insights into the pathogenesis and potential therapeutic targets for migraine.

NOTCH2NLC Intermediate-Length Repeat Expansions Are Associated with Parkinson Disease.

NOTCH2NLC GGC repeat expansions were recently identified in neuronal intranuclear inclusion disease (NIID); however, it remains unclear whether they occur in other neurodegenerative disorders. This study aimed to investigate the role of intermediate-length NOTCH2NLC GGC repeat expansions in Parkinson disease (PD). We screened for GGC repeat expansions in a cohort of 1,011 PD patients and identified 11 patients with intermediate-length repeat expansions ranging from 41 to 52 repeats, with no repeat expansions in 1,134 controls. Skin biopsy revealed phospho-alpha-synuclein deposition, confirming the PD diagnosis in 2 patients harboring intermediate-length repeat expansions instead of NIID or essential tremor. Fibroblasts from PD patients harboring intermediate-length repeat expansions revealed NOTCH2NLC upregulation and autophagic dysfunction. Our results suggest that intermediate-length repeat expansions in NOTCH2NLC are potentially associated with PD. ANN NEUROL 2021;89:182-187.

Null-free False Discovery Rate Control Using Decoy Permutations.

The traditional approaches to false discovery rate (FDR) control in multiple hypothesis testing are usually based on the null distribution of a test statistic. However, all types of null distributions, including the theoretical, permutation-based and empirical ones, have some inherent drawbacks. For example, the theoretical null might fail because of improper assumptions on the sample distribution. Here, we propose a null distribution-free approach to FDR control for multiple hypothesis testing in the case-control study. This approach, named target-decoy procedure, simply builds on the ordering of tests by some statistic or score, the null distribution of which is not required to be known. Competitive decoy tests are constructed from permutations of original samples and are used to estimate the false target discoveries. We prove that this approach controls the FDR when the score function is symmetric and the scores are independent between different tests. Simulation demonstrates that it is more stable and powerful than two popular traditional approaches, even in the existence of dependency. Evaluation is also made on two real datasets, including an arabidopsis genomics dataset and a COVID-19 proteomics dataset.

Novel Single-Enzyme-Assisted Dual Recycle Amplification Strategy for Sensitive Photoelectrochemical MicroRNA Assay.

Herein, a novel single-enzyme-assisted dual recycle amplification strategy based on T7 exonuclease (T7 Exo) and a strand-displacement reaction (SDR) was designed to fabricate a photoelectrochemical (PEC) biosensor for sensitive microRNA-141 (miRNA-141) detection with the use of laminar bismuth tungstate (Bi2WO6) as photoactive material. Compared with a traditional enzyme-assisted dual recycle amplification strategy, the presented method could effectively refrain the enzyme interference reaction, reduce environmental sensitivity, and save cost. Here, hairpin DNA1 (H1) decorated on magnetic beads (MB) hybridized with target miRNA-141 to form an H1/miRNA-141 heteroduplex. With the introduction of hairpin DNA2 (H2)-labeled SiO2 (H2-SiO2), SDR was triggered between H2-SiO2 and H1, thus miRNA-141 was displaced from the H1/miRNA-141 heteroduplex and an H1/H2-SiO2 duplex was formed, realizing the reuse of the target. In the presence of T7 Exo, the H1/H2-SiO2 duplex was digested with the release of output DNA-SiO2. To enhance the target conversion rate, H1-MB was intactly released and cycled, which could initiate more T7 Exo digestion and free abundant output DNA-SiO2. Through such a process, a tiny miRNA-141 could induce substantial output DNA-SiO2, effectively improving the target amplification efficiency and detection sensitivity of a PEC biosensor. Furthermore, Bi2WO6 was modified on an electrode to provide a superior initial PEC signal due to its excellent electronic transformation capacity. With the introduction of output DNA-SiO2, the hairpin structure of H3 on the electrode was opened, making SiO2 close to the electrode surface, which significantly decreases the PEC signal. This work first established the PEC biosensor featuring a single-enzyme-assisted dual recycle amplification process for sensitive detection of biomarkers.

Resveratrol ameliorates thymus senescence changes in D-galactose induced mice.

The thymic microenvironment plays an important role in the development of T cells. A decrease of thymic epithelial cells is the main cause of age-related thymic atrophy or degeneration. Resveratrol (RSV), a phytoalexin produced from plants, has been shown to inhibit the adverse effects of dietary obesity on the structure and function of the thymus. D-Galactose (D-gal) can induce accelerated aging in mice. In the present study, young mice (2 months old) were injected with D-gal (120 mg/kg/day) for 8 consecutive weeks to construct an accelerated aging model. Compared with normal control mice, the thymus epithelium of the D-gal treated mice had structural changes, the number of senescent cells increased, the number of CD4+ T cells decreased, and CD8+ T cells increased. After RSV administration by gavage for 6 weeks, it was found that RSV improved the surface phenotypes of D-gal treated mice, and recovered thymus function by maintaining the ratio of CD4+ to CD8+ cells. It also indicated that RSV enhanced the cell proliferation and inhibited cell senescence. Increased autoimmune regulator (Aire) expression was present in the RSV treated mice. The lymphotoxin-beta receptor (LTßR) expression also increased. These findings suggested that RSV intake could restore the alterations caused by D-gal treatment in the thymus via stimulation of Aire expression.

Supersensitive Photoelectrochemical Aptasensor Based on Br,N-Codoped TiO2 Sensitized by Quantum Dots.

Here, we fabricated a novel photoelectrochemical (PEC) aptasensor based on Br,N-codoped TiO2/CdS quantum dots (QDs) sensitization structure with excellent energy level arrangement for supersensitive detection of carcinoembryonic antigen (CEA). The prepared Br,N-codoped TiO2 could reduce the energy bandwidth of TiO2 from 3.2 to 2.88 eV, which could dramatically reduce the basic signal and obviously broaden the absorption of light (400-700 nm). In addition, the energy bandwidth of Br,N-codoped TiO2 (2.88 eV) matched well with that of CdS QDs (2.4 eV), making CdS QDs an ideal signal enhancer for amplifying the photocurrent signal of Br,N-codoped TiO2. More importantly, the constructed Br,N-codoped TiO2/CdS QDs sensitization structure with narrow energy level gradient enabled the effective promotion of electron-transfer capability and dramatic improvement of photoelectric conversion efficiency. Simultaneously, a small amount of the CEA was transformed into substantial single-chain DNA (T-DNA) via exonuclease III (Exo-III)-assisted cycle strategy. Under optimum conditions, the designed PEC aptasensor demonstrated a wide detection range from 1 fg/mL to 1 ng/mL and a low detection limit as 0.46 fg/mL for CEA assay. This strategy prepared a new photoactive material to markedly improve photoelectric conversion efficiency and initiated a new way to realize the highly sensitive PEC biomolecules detection.

[Ru(dcbpy)2 dppz]2+ /Fullerene Cosensitized PTB7-Th for Ultrasensitive Photoelectrochemical MicroRNA Assay.

A new cosensitization photoelectrochemical (PEC) strategy was established by using a donor-acceptor-type photoactive material, poly{4,8-bis[5-(2-ethylhexyl)thiophen-2-yl]benzo[1,2-b:4,5-b']dithiophene-2,6-diyl-alt-3-fluoro-2-[(2-ethylhexyl)carbonyl]thieno[3,4-b]thiophene-4,6-diyl} (PTB7-Th), as a signal indicator, which was cosensitized with bis(4,4'dicarboxyl-2,2'-bipyridyl)(4,5,9,14-tetraazabenzo[b]triphenylene)ruthenium(II) ([Ru(dcbpy)2 dppz]2+ ) embedded in the grooves of the DNA duplex and fullerene (nano-C60 ) immobilized on the surface of DNA nanoflowers for microRNA assay. [Ru(dcbpy)2 dppz]2+ and nano-C60 could effectively enhance the photoelectric conversion efficiency (PCE) of PTB7-Th as a result of well-matched energy levels among nano-C60 , [Ru(dcbpy)2 dppz]2+ and PTB7-Th, leading to a clearly enhanced photocurrent signal. Meanwhile, a target recycling magnification technique based on duplex-specific nuclease was applied in this work to obtain higher detection sensitivity. The proposed biosensor demonstrated excellent analytical properties within a linear detection range of 2.5 fm to 2.5 nm and a limit of detection down to 0.83 fm. Impressively, this cosensitization PEC strategy offers an effective and convenient avenue to significantly improve the PCE of a photoactive material, resulting in a remarkably improved photocurrent signal for ultrasensitive and highly accurate detection of various targets.

The relationship between psychosocial risk factors, burnout and quality of life among primary healthcare workers in rural Guangdong province: a cross-sectional study.

BACKGROUND: Healthcare workers are often exposed to stressful working conditions at work which affect their quality of life. The study investigated the relationship between psychosocial risk factors, stress, burnout, and quality of life among primary healthcare workers in general medical practice in Qingyuan and Chaozhou cities in Guangdong province. METHOD: The cross-sectional study was conducted in 108 primary health facilities including 36 community health centers (CHCs) across two developing cities in Guangdong province. A total of 873 healthcare workers completed the questionnaires. Quality of life was evaluated using The World Health Organization Quality of Life Questionnaire (WHOQOL-BREF) and psychological risk factors were evaluated by the Copenhagen Psychosocial Questionnaire (COPSOQ). General quality of life and the quality of life domains were transformed into a score range from minimum 0 to 100 maximum. Higher scores indicated better quality of life and vice versa. Significant associations were verified using multiple regression analysis. RESULTS: Poor quality of life was observed in 74.6% of healthcare workers surveyed. General poor quality of life was significantly higher among workers who reported higher burnout (Beta = - 0.331, p < 0.001). In addition, workers with high levels of burnout, unmarried workers and female workers had a higher possibility of physical health. A greater risk of poor psychological health was observed among workers with high burnout, poor sense of community and those with lower educational levels. Workers who lacked social support, those with fewer possibilities for development had increased probability of poor quality of life in the social domain. Poor quality of life in the environmental domain was observed among workers who were dissatisfied with their jobs and workers with low salaries. CONCLUSIONS: Primary healthcare workers in developing cities in China have a highly demanding and strained working environment and poor quality of life. Reducing job stress and improving work conditions may ultimately improve the well-being of primary healthcare workers.

Cosensitization Strategy with Cascade Energy Level Arrangement for Ultrasensitive Photoelectrochemical Protein Detection.

Here, a photoelectrochemical (PEC) biosensor was established by a cosensitization strategy with cascade energy level arrangement for ultrasensitive detection of prostate-specific antigen (PSA). The proposed cosensitization strategy was based on the well-matched energy level arrangement of four kinds of organic photoactive materials, in which poly{4,8-bis[5-(2-ethylhexyl)thiophen-2-yl]benzo[1,2- b:4,5- b']dithiophene-2,6-diyl- alt-3-fluoro-2-[(2-ethylhexyl)-carbonyl]thieno[3,4- b]thiophene-4,6-diyl} (PTB7-Th) was used as the photoactive material and perylenetetracarboxyl diimide (PDI), fullerene (nano-C60), and polyaniline (PANI) were employed as the sensitizers. The resulting PTB7-Th/PDI/nano-C60/PANI cascade cosensitization structure with narrow energy level gradient (<0.54 eV) could effectively improve electron transfer capability, obviously raise light energy utilization and significantly enhance photoelectric conversion efficiency, leading to dramatically enhanced photocurrent response. Using PSA as a target model, the proposed PEC biosensor exhibited high sensitivity and excellent stability with a wide detection range from 1 fg/mL to 0.1 ng/mL and a detection limit of 0.43 fg/mL. Moreover, the proposed PEC biosensor provides a cascade cosensitization strategy that could significantly improve PEC performances and open up a promising platform to establish high selectivity, stability, and ultrasensitive analytical techniques.

A Highly Sensitive Photoelectrochemical Assay with Donor-Acceptor-Type Material as Photoactive Material and Polyaniline as Signal Enhancer.

In this work, a highly sensitive photoelectrochemical (PEC) assay was constructed based on a donor-acceptor (D-A)-type material, poly{4,8-bis[5-(2-ethylhexyl)thiophen-2-yl]benzo[1,2-b:4,5-b']dithiophene-2,6-diyl- alt-3-fluoro-2-[(2-ethylhexyl)-carbonyl]thieno[3,4- b]thiophene-4,6-diyl} (PTB7-Th), as the photoactive material and polyaniline (PANI) in situ deposited on the surface of PTB7-Th as the signal enhancer. Initially, PTB7-Th, which contains an electron-rich unit as donor and an electron-deficient unit as acceptor with an easy separation of electron-hole pairs and intermolecular electron transfer, provided an excellent photocurrent response. Subsequently, an input target thrombin (TB) was converted to an output single-stranded DNA by a protein converting strategy. The obtained single-stranded DNA thus triggered a rolling circle amplification (RCA) to form a tandem multihairpin DNA nanostructure, which could function as a skeleton for immobilizing manganese porphyrin (MnTMPyP). In the presence of H2O2 and aniline, a PANI layer could be in situ deposited onto the tandem multihairpin DNA nanostructure with use of MnTMPyP as catalyst, leading to a significantly enhanced photocurrent for the detection of TB. The proposed PEC assay presented a wide detection range of 100 fM to 10 nM with a limit of detection (LOD) of 34.6 fM. Furthermore, the proposed strategy provides a PEC analysis method based on PTB7-Th that can significantly improve the photoelectric conversion efficiency and opens an intriguing avenue to establish low background, ultrasensitive, and highly stable analytical techniques.

Combined effects of a high-fat diet and chronic valproic acid treatment on hepatic steatosis and hepatotoxicity in rats.

AIM: To investigate the potential interactive effects of a high-fat diet (HFD) and valproic acid (VPA) on hepatic steatosis and hepatotoxicity in rats. METHODS: Male SD rats were orally administered VPA (100 or 500 mg·kg⁻¹·d⁻¹) combined with HFD or a standard diet for 8 weeks. Blood and liver samples were analyzed to determine lipid levels and hepatic function biomarkers using commercial kit assays. Low-molecular-weight compounds in serum, urine and bile samples were analyzed using a metabonomic approach based on GC/TOF-MS. RESULTS: HFD alone induced extensive hepatocyte steatosis and edema in rats, while VPA alone did not cause significant liver lesions. VPA significantly aggravated HFD-induced accumulation of liver lipids, and caused additional spotty or piecemeal necrosis, accompanied by moderate infiltration of inflammatory cells in the liver. Metabonomic analysis of serum, urine and bile samples revealed that HFD significantly increased the levels of amino acids, free fatty acids (FFAs) and 3-hydroxy-butanoic acid, whereas VPA markedly decreased the levels of amino acids, FFAs and the intermediate products of the tricarboxylic acid cycle (TCA) compared with the control group. HFD aggravated VPA-induced inhibition on lipid and amino acid metabolism. CONCLUSION: HFD magnifies VPA-induced impairment of mitochondrial ß-oxidation of FFAs and TCA, thereby increases hepatic steatosis and hepatotoxicity. The results suggest the patients receiving VPA treatment should be advised to avoid eating HFD.

A comparison between the pectoralis major myocutaneous flap and the free anterolateral thigh perforator flap for reconstruction in head and neck cancer patients: assessment of the quality of life.

Our study investigated the quality of life (QoL) of Chinese patients after immediate reconstruction surgery on individuals with head and neck cancer. In addition, we compared the differences between pectoralis major myocutaneous flap (PMMF) and anterolateral thigh free flap (ALTFF). The University of Washington Quality of Life questionnaire, version 4, was used to assess the QoL. Assessments were performed at least 24 months postoperatively. A total of 110 patients' records were obtained. Among them, 86 patients completed a QoL questionnaire (78.2%). No significant differences could be found in age, primary site, T stage, N stage, and postoperative radiotherapy between PMMF and ALTFF groups. However, there were significant differences between both groups in sex, operation time, and complication. A matched analysis was performed to compare the differences in QoL between patients with head and neck cancers reconstructed with PMMF or ALTFF. Patients reconstructed with ALTFF had better shoulder but worse speech functions. There was a significant effect on the QoL of head and neck cancer patients who had undergone either PMMF or ALTFF reconstruction. The result of this study provide useful information for physicians and patients during their discussion of treatment modalities for head and neck cancers.

Metagenome next-generation sequencing plays a key role in the diagnosis and selection of effective antibiotics on the treatment of Nocardia pneumonia: a case report.

Nocardia disease is an opportunistic infection, the occurrence is rare and mostly occurs in patients with immune deficiency. Even if the patient is immunocompetent, it can still be life-threatening. This case report describes a previously healthy 78-year-old male farmer with lung lesions discovered on a computerized tomography scan. Combined with the patient's history of fever and the results of elevated laboratory markers associated with inflammation, the patient was diagnosed with a lung infection. After escalating empirical broad-spectrum antibiotics, antiviral and antifungal therapy, the patient continued to deteriorate to septic shock. In the meanwhile, the patient's sputum was cultured repeatedly, and no obvious positive pathogenic bacteria were found. Considering the patient was elderly and that these lesions were solid with burr signs, as well as the progression after antimicrobial therapy cancer was considered in the differential diagnosis. Artificial intelligence (YITU, Hangzhou Yitu Medical Technology Limited Company) was also applied, and it also calculated that these lesions were cancerous. The patient received a puncture biopsy of the largest lung lesion. During the puncture pus was withdrawn from largest lung lesion. Culture and metagenome next-generation sequencing (mNGS) detection performed on pus indicated Nocardia otitidiscaviarum. The test report of the mNGS is also attached with a susceptibility report of commonly used clinical antibiotics to this Nocardia spp. Using this result, the patient's disease was quickly controlled after selecting the targeted drug compound sulfamethoxazole and intravenous meropenem for treatment. In view of the high misdiagnosis rate and poor sensitivity of culture for Nocardia spp., this case emphasized mNGS playing a key role in the diagnosis and selection of effective antibiotics for the treatment of Nocardia spp. lung infections.

Peptidoglycan biosynthesis-associated enzymatic kinetic characteristics and ß-lactam antibiotic inhibitory effects of different Streptococcus pneumoniae penicillin-binding proteins.

Penicillin-binding proteins (PBPs) include transpeptidases, carboxypeptidases, and endopeptidases for biosynthesis of peptidoglycans in the cell wall to maintain bacterial morphology and survival in the environment. Streptococcus pneumoniae expresses six PBPs, but their enzymatic kinetic characteristics and inhibitory effects on different ß-lactam antibiotics remain poorly understood. In this study, all the six recombinant PBPs of S. pneumoniae displayed transpeptidase activity with different substrate affinities (Km = 1.56-9.11 mM) in a concentration-dependent manner, and rPBP3 showed a greater catalytic efficiency (Kcat = 2.38 s-1) than the other rPBPs (Kcat = 3.20-7.49 × 10-2 s-1). However, only rPBP3 was identified as a carboxypeptidase (Km = 8.57 mM and Kcat = 2.57 s-1). None of the rPBPs exhibited endopeptidase activity. Penicillin and cefotaxime inhibited the transpeptidase and carboxypeptidase activity of all the rPBPs but imipenem did not inhibited the enzymatic activities of rPBP3. Except for the lack of binding of imipenem to rPBP3, penicillin, cefotaxime, and imipenem bound to all the other rPBPs (KD = 3.71-9.35 × 10-4 M). Sublethal concentrations of penicillin, cefotaxime, and imipenem induced a decrease of pneumococcal pbps-mRNA levels (p < 0.05). These results indicated that all six PBPs of S. pneumoniae are transpeptidases, while only PBP3 is a carboxypeptidase. Imipenem has no inhibitory effect on pneumococcal PBP3. The pneumococcal genes for encoding endopeptidases remain to be determined.

The potential protective role of peripheral immunophenotypes in Alzheimer's disease: a Mendelian randomization study.

Introduction: Alzheimer's disease (AD) is the most widespread neurodegenerative disease in the world. Previous studies have shown that peripheral immune dysregulation plays a paramount role in AD, but whether there is a protective causal relationship between peripheral immunophenotypes and AD risk remains ambiguous. Methods: Two-sample Mendelian randomization (MR) was performed using large genome-wide association study (GWAS) genetic data to assess causal effects between peripheral immunophenotypes and AD risk. Utilizing the genetic associations of 731 immune cell traits as exposures. We adopted the inverse variance weighted method as the primary approach. The Weighted median and MR-Egger regression methods were employed as supplements. Various sensitivity analyses were performed to assess the robustness of the outcomes. Results: Based on the IVW method, we identified 14 immune cell traits that significantly reduced the risk of AD, of which six demonstrated statistical significance in both IVW and Weighted median methods. Among the seven immune traits, four were related to regulatory T (Treg) cells : (1) CD25++ CD45RA- CD4 not regulatory T cell % T cell (odds ratio (OR) [95% confidence interval (CI)] = 0.96 [0.95, 0.98], adjusted P = 1.17E-02), (2) CD25++ CD45RA- CD4 not regulatory T cell % CD4+ T cell (OR [95% CI] = 0.97 [0.96, 0.99], adjusted P = 3.77E-02), (3) Secreting CD4 regulatory T cell % CD4 regulatory T cell (OR [95% CI] = 0.98 [0.97, 0.99], adjusted P = 7.10E-03), (4) Activated & secreting CD4 regulatory T cell % CD4 regulatory T cell(OR [95% CI] = 0.98 [0.97, 0.99], adjusted P = 7.10E-03). In addition, HLA DR++ monocyte % monocyte (OR [95% CI] = 0.93 [0.89, 0.98], adjusted P = 4.87E-02) was associated with monocytes, and HLA DR on myeloid Dendritic Cell (OR [95% CI] = 0.93 [0.89, 0.97], adjusted P = 1.17E-02) was related to dendritic cells (DCs). Conclusion: These findings enhance the comprehension of the protective role of peripheral immunity in AD and provide further support for Treg and monocyte as potential targets for immunotherapy in AD.

Joint analysis of proteome, transcriptome, and multi-trait analysis to identify novel Parkinson's disease risk genes.

Genome-wide association studies (GWAS) have identified multiple risk variants for Parkinson's disease (PD). Nevertheless, how the risk variants confer the risk of PD remains largely unknown. We conducted a proteome-wide association study (PWAS) and summary-data-based mendelian randomization (SMR) analysis by integrating PD GWAS with proteome and protein quantitative trait loci (pQTL) data from human brain, plasma and CSF. We also performed a large transcriptome-wide association study (TWAS) and Fine-mapping of causal gene sets (FOCUS), leveraging joint-tissue imputation (JTI) prediction models of 22 tissues to identify and prioritize putatively causal genes. We further conducted PWAS, SMR, TWAS, and FOCUS using a multi-trait analysis of GWAS (MTAG) to identify additional PD risk genes to boost statistical power. In this large-scale study, we identified 16 genes whose genetically regulated protein abundance levels were associated with Parkinson's disease risk. We undertook a large-scale analysis of PD and correlated traits, through TWAS and FOCUS studies, and discovered 26 casual genes related to PD that had not been reported in previous TWAS. 5 genes (CD38, GPNMB, RAB29, TMEM175, TTC19) showed significant associations with PD at both the proteome-wide and transcriptome-wide levels. Our study provides new insights into the etiology and underlying genetic architecture of PD.