Bioenergetic stress potentiates antimicrobial resistance and persistence.

Antimicrobial resistance (AMR) is a global health crisis and there is an urgent need to better understand AMR mechanisms. Antibiotic treatment alters several aspects of bacterial physiology, including increased ATP utilization, carbon metabolism, and reactive oxygen species (ROS) formation. However, how the "bioenergetic stress" induced by increased ATP utilization affects treatment outcomes is unknown. Here we utilized a synthetic biology approach to study the direct effects of bioenergetic stress on antibiotic efficacy. We engineered a genetic system that constitutively hydrolyzes ATP or NADH in Escherichia coli. We found that bioenergetic stress potentiates AMR evolution via enhanced ROS production, mutagenic break repair, and transcription-coupled repair. We also find that bioenergetic stress potentiates antimicrobial persistence via potentiated stringent response activation. We propose a unifying model that antibiotic-induced antimicrobial resistance and persistence is caused by antibiotic-induced. This has important implications for preventing or curbing the spread of AMR infections.

[The Development of SpCas9 Variants with High Specificity and Efficiency Based on the HH Theory].

Streptococcus pyogenes Cas9 (SpCas9) is the most popular tool in gene editing; however, off-target mutagenesis is one of the biggest impediments in its application. In our previous study, we proposed the HH theory, which states that sgRNA/DNA hybrid (hybrid) extrusion-induced enhancement of hydrophobic interactions between the hybrid and REC3/HNH is a key factor in cleavage initiation. Based on the HH theory, we analyzed the interactions between the REC3 domain and hybrid and obtained 8 mutant sites. We designed 8 SpCas9 variants (V1-V8), used digital droplet PCR to assess SpCas9-induced DNA indels in human cells, and developed high-fidelity variants. Thus, the HH theory may be employed to further optimize SpCas9-mediated genome editing systems, and the resultant V3, V6, V7, and V8 SpCas9 variants may be valuable for applications requiring high-precision genome editing.

DNA Repair Genetics and the Risk of Radiation Pneumonitis in Patients With Lung Cancer: A Systematic Review and Meta-analysis.

AIMS: ERCC1 rs11615 and ERCC2 rs238406 single nuclear polymorphism (SNPs) are known for their association with treatment outcome, likely related to radiosensitivity of both tumor and normal tissue in patients with non-small-cell lung cancer. This study aimed to review the effect of 1) these ERCC1/2 SNPs and 2) other SNPs of DNA repair genes on radiation pneumonitis (RP) in patients with lung cancer. MATERIALS AND METHODS: SNPs of our interest included ERCC1 rs11615 and ERCC2 rs238406 and other genes of DNA repair pathways that are functional and biologically active. DNA repair SNPs reported by at least two independent studies were pooled for meta-analysis. The study endpoint was radiation pneumonitis (RP) after radiotherapy. Recessive, dominant, homozygous, heterozygous, and allelic genotype models were used where appropriate. RESULTS: A total of 16 studies (3080 patients) were identified from the systematic review and 12 studies (2090 patients) on 11 SNPs were included in the meta-analysis. The SNPs were ATM rs189037, ATM rs373759, NEIL1 rs4462560, NEIL1 rs7402844, APE1 rs1130409, XRCC3 rs861539, ERCC1 rs11615, ERCC1 rs3212986, ERCC2 rs238406, ERCC2 rs13181, and XRCC1 rs25487. ERCC1 rs11615 (236 patients) and ERCC2 rs238406 (254 patients) were not significantly associated with RP. Using the allelic model, the G allele for NEIL1 gene was significantly associated with a reduced odds of developing symptomatic (grade ≥2) RP compared to the C allele for rs7402844 (OR 0.70, 95% CI: 0.49, 0.99, P = 0.04). Similarly, the T allele for APE1 gene was significantly associated with a reduced odds of developing symptomatic (grade ≥2) RP compared to the G allele for rs1130409 (OR 0.59, 95% CI: 0.43, 0.81, P = 0.001). CONCLUSION: Genetic variation in the DNA repair pathway genes may play a significant role in the risk of developing radiation pneumonitis in patients with lung cancer. Further studies are needed on genotypic features of DNA repair pathway genes and their association with treatment sensitivity, as such knowledge may guide personalized radiation dose prescription.

Cytidine Acetylation Across the Tree of Life.

Acetylation plays a critical role in regulating eukaryotic transcription via the modification of histones. Beyond this well-documented function, a less explored biological frontier is the potential for acetylation to modify and regulate the function of RNA molecules themselves. N4-Acetylcytdine (ac4C) is a minor RNA nucleobase conserved across all three domains of life (archaea, bacteria, and eukarya), a conservation that suggests a fundamental role in biological processes. Unlike many RNA modifications that are controlled by large enzyme families, almost all organisms catalyze ac4C using a homologue of human Nat10, an essential disease-associated acetyltransferase enzyme.A critical step in defining the fundamental functions of RNA modifications has been the development of methods for their sensitive and specific detection. This Account describes recent progress enabling the use of chemical sequencing reactions to map and quantify ac4C with single-nucleotide resolution in RNA. To orient readers, we first provide historical background of the discovery of ac4C and the enzymes that catalyze its formation. Next, we describe mechanistic experiments that led to the development of first- and second-generation sequencing reactions able to determine ac4C's position in a polynucleotide by exploiting the nucleobase's selective susceptibility to reduction by hydride donors. A notable feature of this chemistry, which may serve as a prototype for nucleotide resolution RNA modification sequencing reactions more broadly, is its ability to drive a penetrant and detectable gain of signal specifically at ac4C sites. Emphasizing practical applications, we present how this optimized chemistry can be integrated into experimental workflows capable of sensitive, transcriptome-wide analysis. Such readouts can be applied to quantitatively define the ac4C landscape across the tree of life. For example, in human cell lines and yeast, this method has uncovered that ac4C is highly selective, predominantly occupying dominant sites within rRNA (rRNA) and tRNA (tRNA). By contrast, when we extend these analyses to thermophilic archaea they identify the potential for much more prevalent patterns of cytidine acetylation, leading to the discovery of a role for this modification in adaptation to environmental stress. Nucleotide resolution analyses of ac4C have also allowed for the determination of structure-activity relationships required for short nucleolar RNA (snoRNA)-catalyzed ac4C deposition and the discovery of organisms with unexpectedly divergent tRNA and rRNA acetylation signatures. Finally, we share how these studies have shaped our approach to evaluating novel ac4C sites reported in the literature and highlight unanswered questions and new directions that set the stage for future research in the field.

Genetic parameters for early piglet weight, litter traits and number of functional teats in organic pigs.

Knowledge remains limited on genetic variation and genetic correlations for traits in sows and piglets that are reared in an organic or outdoor setting. Here, we estimated genetic variance components for individual piglet weight, litter weight, litter size traits, and number of functional teats in a pig population raised under outdoor organic conditions. Data were collected from the largest organic multiplier farm in Denmark. Individual piglet weight was recorded at birth and on day 10. Number of live and dead piglets were recorded at birth, day 4, and day 11. Mean and total litter weight were calculated based on the individual weight of living piglets at birth and on day 10. The estimated heritability was highest for the number of functional teats (0.49), mean weight of a litter at birth (0.33) and on day 10 (0.25). In contrast, heritability was lowest for litter size traits (0.04-0.08) and piglet weight (0.06-0.07). Maternal heritability was much higher for individual piglet weight than direct heritability. The results showed that selection for higher mean weight results in smaller litters. Also, selection for individual birth weight of piglets results in heavier piglets at 10 days. In conclusion, this study confirmed that there is genetic variation in individual piglet weight, litter traits, and number of functional teats in organically and outdoor-reared pigs.

The Relationship between Psychological Distress and Physical Frailty in Japanese Community-Dwelling Older Adults: A Cross-Sectional Study.

BACKGROUND: Older adults' mental health and physical frailty have been a frequent research focus, but few studies have investigated the relationship between them. OBJECTIVES: To investigate the association between mental health and physical frailty in community-dwelling older Japanese people. DESIGN: Cross-sectional study from the Itoshima Frail Study. SETTING: Itoshima City, Fukuoka, Japan. PARTICIPANTS: A total of 919 community-dwelling older individuals aged 65-75 years. MEASUREMENTS: Physical frailty was measured based on five criteria proposed by the Fried scale, and the subjects were classified into three groups: robust, pre-frailty, and frailty. Psychological distress was used to assess the subjects' mental health, with the Kessler 6-Item Psychological Distress Scale (K6) score; the subjects were divided into three groups based on their K6 score: 0-1, 2-4, and ≥5. Psychological distress was defined by K6 score ≥5. Ordinal logistic regression was used to estimate the odds ratios (OR) and 95% confidence intervals (CIs) between the psychological distress and physical frailty status. RESULTS: Psychological distress was identified in 190 subjects (20.7%). Forty-six subjects (5.0%) presented with physical frailty, and 24 subjects (2.6%) had both psychological distress and physical frailty. With the increase in the K6 score, more subjects had pre-frailty and physical frailty (p<0.001). Significant positive associations were observed between psychological distress and the risks of pre-frailty (OR 2.94, 95%CI: 1.95-4.43) and frailty (OR 10.71, 95%CI: 4.68-24.51), even in a multivariable-adjusted analysis. In a subgroup analysis of components of frailty, one-point increment in K6 score was associated with higher odds of shrinking and fatigue. CONCLUSION: A severe psychological distress was associated with increased risks of physical frailty and the frailty sub-items of shrinking and fatigue in community-dwelling older Japanese adults.

[The influence of various myelosuppression degrees during neoadjuvant chemotherapy on the curative effect and prognosis of triple-negative breast cancer].

Objective: To investigate the effect of the degrees of myelosuppression on the curative effect and prognosis of triple-negative breast cancer with neoadjuvant chemotherapy. Methods: The clinical, pathological and follow-up data of 206 patients with triple negative breast cancer who received neoadjuvant chemotherapy with docetaxel combined with epirubicin combined with cyclophosphamide regimen in the Department of Breast Surgery in the Third Affiliated Hospital of Zhengzhou University from January 2013 to December 2018 were collected retrospectively. All were female, aged 28-71 (47.8±10.7) years. According to the WHO classification standard of acute and subacute toxicity of anticancer drugs, the patients were divided into 98 cases in the mild group (0-Ⅱ degree) and 108 cases in the severe group (Ⅲ-Ⅳ degree) according to the degree of bone marrow suppression after chemotherapy. The baseline clinicopathological features, pathological complete remission rate (PCR) and objective remission rate (ORR) of the two groups were compared. The survival curve was drawn by Kaplan Meier method, and the differences of disease-free survival (DFS), local recurrence free survival (LRFS), distant metastasis free survival (DMFS) and overall survival (OS) between the two groups were analyzed by log rank test. Cox regression risk model was used to analyze the related factors affecting the survival of the patients. Results: There were no significant differences in baseline clinicopathological characteristics of patients between the two groups, such as age, physical status score, menopausal status, body mass index, histological grade, clinical T stage, clinical N stage and Ki-67 index (all P>0.05). The severe group had higher PCR, longer median DFS and median DMFS than the mild group [50.9%(55/108) vs 36.7%(36/98); not reached vs 72 months; not reached vs 84 months] (all P<0.05). There was no significant difference in ORR, LRFS and OS between the two groups [89.8%(97/108) vs 81.6%(80/98); both not reached; both not reached] (all P>0.05). The degree of bone marrow suppression after neoadjuvant chemotherapy was an influential factor of DFS in TNBC patients (P=0.025). Compared with mild myelosuppression group, severe myelosuppression group had better disease-free survival prognosis (HR=0.571, 95%CI: 0.349-0.934). Conclusion: The prognosis of grade Ⅲ/Ⅳ myelosuppression is better than grade 0/Ⅰ/Ⅱ myelosuppression in patients with triple-negative breast cancer during neoadjuvant chemotherapy with TEC regimen, which is helpful for judging efficacy.

Equations of motion for weakly compressible point vortices.

Equations of motion for compressible point vortices in the plane are obtained in the limit of small Mach number, M, using a Rayleigh-Jansen expansion and the method of Matched Asymptotic Expansions. The solution in the region between vortices is matched to solutions around each vortex core. The motion of the vortices is modified over long time scales [Formula: see text] and [Formula: see text]. Examples are given for co-rotating and co-propagating vortex pairs. The former show a correction to the rotation rate and, in general, to the centre and radius of rotation, while the latter recover the known result that the steady propagation velocity is unchanged. For unsteady configurations, the vortex solution matches to a far field in which acoustic waves are radiated. This article is part of the theme issue 'Mathematical problems in physical fluid dynamics (part 2)'.

Safety, pharmacokinetics, and efficacy of BPI-15086 in patients with EGFR T790M-mutated advanced non-small-cell lung cancer: results from a phase I, single-arm, multicenter study.

BACKGROUND: Epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (TKI) resistance frequently occurs in patients with non-small-cell lung cancer (NSCLC). EGFR Thr790Met mutation (T790M+) is seen in ∼50% of patients. We assessed the safety, tolerability, and pharmacokinetics (PK) of BPI-15086, a novel, ATP-competitive, irreversible, third-generation, mutation-selective EGFR-TKI in patients with EGFR T790M-mutated NSCLC. PATIENTS AND METHODS: This two-center, phase I, dose-escalation study included patients who were 18-65 years old, with an Eastern Cooperative Oncology Group performance status of 0-2, with histologically or cytologically confirmed locally advanced or metastatic T790M+ NSCLC who were not surgical or radiotherapy candidates, and had imaging-identified disease progression after prior EGFR-TKIs. This dose-escalation study enrolled patients using a 3 + 3 study design. Patients received 25, 50, 100, 200, and 300 mg/day orally in 21-day cycles. The primary endpoints were safety, tolerability, and PK. Secondary endpoints were objective response rate (ORR) and disease control rate (DCR). The dose-expansion study was not conducted. RESULTS: We enrolled 17 patients from 29 December 2016 to 16 May 2018, in the safety and full analysis sets. All patients completed a single dosing trial, and no adverse events (AEs) causing drug discontinuation were seen. Grade 1-2 nausea, hypoalbuminemia, and decreased appetite were the most common treatment-related AEs. Grade 3 hyperglycemia was seen in one patient dosed at 300 mg/day. The ORR and DCR were 17.7% [95% confidence interval (CI) 3.8% to 43.4%] and 47.1% (95% CI 23.0% to 72.2%), respectively. CONCLUSION: BPI-15086 is a safe and tolerable third-generation EGFR-TKI with a rationale for further clinical studies.

[Impact of neoadjuvant immunotherapy on pulmonary function and perioperative outcomes in patients with resectable non-small cell lung cancer].

Objective: To explore the effect of neoadjuvant immunotherapy on pulmonary function and the efficacy in patients with resectable non-small cell lung cancer. Methods: Data of 30 patients with non-small cell lung cancer (NSCLC) who received neoadjuvant immunotherapy before surgery in the Chest Hospital of Shanghai Jiaotong University from March 2018 to September 2021 were retrospectively collect. The efficacy and safety of neoadjuvant immunotherapy in the perioperative period and changes in pulmonary function of patients before and after neoadjuvant treatment were valuated. Results: The patients were all-male with age of (61±8)years old, The major pathological response (MPR) rate of patients receiving neoadjuvant immunotherapy was 43%(13 cases), the pathologic complete response (pCR) rate was 37% (11 cases), disease control rate (DCR) was 97% (29 cases), objective response rate (ORR) was 67% (20 cases). The forced expiratory volume in one second (FEV1) after treatment was (2.59±0.63) L, and the ratio of FEV1 to the predicted value (FEV1%pred) was 85.27%±15.86%, which were significantly higher than those before treatment [(2.48±0.59)L, 81.73%±15.94%, respectively] (P=0.013, 0.022, respectively). Forced vital capacity (FVC) after treatment was (3.59±0.77) L, which was also significantly higher than before [(3.47±0.76) L,P=0.036]; while there were no statistical difference in FEV1/FVC and FVC accounted for the proportion of predicted values (FVC%pred) between before and after treatment (P=0.084, 0.344, respectively). The ratio of carbon monoxide dispersion (DLCO) to the predicted value (DLCO%pred) decreased from 83.61%±13.10% to 78.69%±13.85% after treatment (P=0.023). There was no significant difference in the incidence of postoperative complications between the DLCO%pred decreased group and the non-decreased group (3/18 vs 0/6; P=0.546). Conclusions: Neoadjuvant immunotherapy can increase the rate of MPR and PCR, significantly increase FEV1 and FEV1%pred, but also lead to a decrease in DLCO%pred; neoadjuvant immunotherapy does not increase the incidence of postoperative complications.

Two-Photon, Ratiometric, Quantitative Fluorescent Probe Reveals Fluctuation of Peroxynitrite Regulated by Arginase 1.

Peroxynitrite, a transient reactive oxygen species (ROS), is believed to play a deleterious role in physiological processes. Herein, we report a two-photon ratiometric fluorescent probe that selectively reacts with peroxynitrite yielding a >200-fold change upon reaction. The probe effectively visualized fluctuations in peroxynitrite generation by arginase 1 in vivo and in vitro. This provides evidence that arginase 1 is a critical regulator of peroxynitrite.

High-Throughput miRFluR Platform Identifies miRNA Regulating B3GLCT That Predict Peters' Plus Syndrome Phenotype, Supporting the miRNA Proxy Hypothesis.

MicroRNAs (miRNAs, miRs) finely tune protein expression and target networks of hundreds to thousands of genes that control specific biological processes. They are critical regulators of glycosylation, one of the most diverse and abundant post-translational modifications. In recent work, miRs have been shown to predict the biological functions of glycosylation enzymes, leading to the "miRNA proxy hypothesis" which states, "if a miR drives a specific biological phenotype..., the targets of that miR will drive the same biological phenotype." Testing of this powerful hypothesis is hampered by our lack of knowledge about miR targets. Target prediction suffers from low accuracy and a high false prediction rate. Herein, we develop a high-throughput experimental platform to analyze miR-target interactions, miRFluR. We utilize this system to analyze the interactions of the entire human miRome with beta-3-glucosyltransferase (B3GLCT), a glycosylation enzyme whose loss underpins the congenital disorder Peters' Plus Syndrome. Although this enzyme is predicted by multiple algorithms to be highly targeted by miRs, we identify only 27 miRs that downregulate B3GLCT, a >96% false positive rate for prediction. Functional enrichment analysis of these validated miRs predicts phenotypes associated with Peters' Plus Syndrome, although B3GLCT is not in their known target network. Thus, biological phenotypes driven by B3GLCT may be driven by the target networks of miRs that regulate this enzyme, providing additional evidence for the miRNA proxy hypothesis.

LncRNA-TMPO-AS1 promotes apoptosis of osteosarcoma cells by targeting miR-329 and regulating E2F1.

OBJECTIVE: This study aims to explore the clinical value and mechanism of lncRNA-TMPO-AS1 in osteosarcoma. PATIENTS AND METHODS: We collected 51 samples of cancer tissues and 51 samples of matched adjacent tissues from 51 patients with osteosarcoma undergoing surgery in our hospital from February 2018 to February 2019. The expression of TMPO-AS1 in tissue samples was tested to analyze its value in the diagnosis and prognosis prediction of osteosarcoma. We transfected osteosarcoma cells with stable and transient vectors containing overexpressed or inhibited genes. Then, we measured cell proliferation by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay, cell invasion by transwell assay, and cell apoptosis by flow cytometry (FCM). The relationship between TMPO-AS1 and miR-329 or between miR-329 and E2F1 was determined by the Dual-Luciferase reporter (DLR) assay. RESULTS: TMPO-AS1 was up-regulated in osteosarcoma cells. Serum TMPO-AS1 could work as a diagnostic marker for osteosarcoma, with an area under the receiver operating characteristic (ROC) curve (AUC) of more than 0.8. Osteosarcoma cells were transfected with siRNA-TMPO-AS1, pcDNA3.1-TMPO-AS1, miR-329-mimics, and miR-329-inhibitor. The results revealed that inhibited TMPO-AS1/overexpressed miR-329/inhibited E2F1 suppressed the proliferation and invasion of osteosarcoma cells and enhanced cell apoptosis. The DLR assay demonstrated that TMPO-AS1 could target miR-329 and miR-329 could target E2F1. In vitro experiments revealed that TMPO-AS1 could regulate the progression of epithelial-mesenchymal transition (EMT) in osteosarcoma through the miR-329/E2F1 axis. CONCLUSIONS: TMPO-AS1 can function as a diagnostic and prognostic marker for osteosarcoma. LncRNA-TMPO-AS1 can promote apoptosis of osteosarcoma cells by targeting miR-329 and regulating E2F1, which is a potent treatment option for osteosarcoma.

Sweet Control: MicroRNA Regulation of the Glycome.

Glycosylation is a sophisticated informational system that controls specific biological functions at the cellular and organismal level. Dysregulation of glycosylation may underlie some of the most complex and common diseases of the modern era. In the past 5 years, microRNAs have come to the forefront as a critical regulator of the glycome. Herein, we review the current literature on miRNA regulation of glycosylation and how this work may point to a new way to identify the biological importance of glycosylation enzymes.

The 100 most cited manuscripts in head and neck cancer: a bibliometric analysis.

OBJECTIVE: This study aimed to highlight the key studies that have led to the current understanding and treatment of head and neck cancer. METHOD: The Thomson Reuters Web of Science database was used to identify relevant manuscripts. The results were ranked according to the number of citations. The 100 most cited papers were analysed. RESULTS: A total of 63 538 eligible papers were returned. The median number of citations was 626. The most cited paper compared radiotherapy with and without cetuximab (3205 citations). The New England Journal of Medicine had the most citations (23 514), and the USA had the greatest number of publications (n = 66). The most common topics of publication were the treatment (n = 45) and basic science (n = 19) of head and neck cancer, followed by the role of human papillomavirus (n = 16). CONCLUSION: This analysis highlighted key articles that influenced head and neck cancer research and treatment. It serves as a guide as to what makes a 'citable' paper in this field.

Testicular tissue cryopreservation: 8 years of experience from a coordinated network of academic centers.

STUDY QUESTION: Is it feasible to disseminate testicular tissue cryopreservation with a standardized protocol through a coordinated network of centers and provide centralized processing/freezing for centers that do not have those capabilities? SUMMARY ANSWER: Centralized processing and freezing of testicular tissue from multiple sites is feasible and accelerates recruitment, providing the statistical power to make inferences that may inform fertility preservation practice. WHAT IS KNOWN ALREADY: Several centers in the USA and abroad are preserving testicular biopsies for patients who cannot preserve sperm in anticipation that cell- or tissue-based therapies can be used in the future to generate sperm and offspring. STUDY DESIGN, SIZE, DURATION: Testicular tissue samples from 189 patients were cryopreserved between January 2011 and November 2018. Medical diagnosis, previous chemotherapy exposure, tissue weight, and presence of germ cells were recorded. PARTICIPANTS/MATERIALS, SETTING, METHODS: Human testicular tissue samples were obtained from patients undergoing treatments likely to cause infertility. Twenty five percent of the patient's tissue was donated to research and 75% was stored for patient's future use. The tissue was weighed, and research tissue was fixed for histological analysis with Periodic acid-Schiff hematoxylin staining and/or immunofluorescence staining for DEAD-box helicase 4, and/or undifferentiated embryonic cell transcription factor 1. MAIN RESULTS AND THE ROLE OF CHANCE: The average age of fertility preservation patients was 7.9 (SD = 5) years and ranged from 5 months to 34 years. The average amount of tissue collected was 411.3 (SD = 837.3) mg and ranged from 14.4 mg-6880.2 mg. Malignancies (n = 118) were the most common indication for testicular tissue freezing, followed by blood disorders (n = 45) and other conditions (n = 26). Thirty nine percent (n = 74) of patients had initiated their chemotherapy prior to undergoing testicular biopsy. Of the 189 patients recruited to date, 137 have been analyzed for the presence of germ cells and germ cells were confirmed in 132. LIMITATIONS, REASONS FOR CAUTION: This is a descriptive study of testicular tissues obtained from patients who were at risk of infertility. The function of spermatogonia in those biopsies could not be tested by transplantation due limited sample size. WIDER IMPLICATIONS OF THE FINDINGS: Patients and/or guardians are willing to pursue an experimental fertility preservation procedure when no alternatives are available. Our coordinated network of centers found that many patients request fertility preservation after initiating gonadotoxic therapies. This study demonstrates that undifferentiated stem and progenitor spermatogonia may be recovered from the testicular tissues of patients who are in the early stages of their treatment and have not yet received an ablative dose of therapy. The function of those spermatogonia was not tested. STUDY FUNDING/COMPETING INTEREST(S): Support for the research was from the Eunice Kennedy Shriver National Institute for Child Health and Human Development grants HD061289 and HD092084, the Scaife Foundation, the Richard King Mellon Foundation, the Departments of Ob/Gyn & Reproductive Sciences and Urology of the University of Pittsburgh Medical Center, United States-Israel Binational Science Foundation (BSF), and the Kahn Foundation. The authors declare that they do not have competing financial interests.

Curcumin restores innate immune Alzheimer's disease risk gene expression to ameliorate Alzheimer pathogenesis.

Alzheimer's disease (AD) genetics implies a causal role for innate immune genes, TREM2 and CD33, products that oppose each other in the downstream Syk tyrosine kinase pathway, activating microglial phagocytosis of amyloid (Aß). We report effects of low (Curc-lo) and high (Curc-hi) doses of curcumin on neuroinflammation in APPsw transgenic mice. Results showed that Curc-lo decreased CD33 and increased TREM2 expression (predicted to decrease AD risk) and also increased TyroBP, which controls a neuroinflammatory gene network implicated in AD as well as phagocytosis markers CD68 and Arg1. Curc-lo coordinately restored tightly correlated relationships between these genes' expression levels, and decreased expression of genes characteristic of toxic pro-inflammatory M1 microglia (CD11b, iNOS, COX-2, IL1ß). In contrast, very high dose curcumin did not show these effects, failed to clear amyloid plaques, and dysregulated gene expression relationships. Curc-lo stimulated microglial migration to and phagocytosis of amyloid plaques both in vivo and in ex vivo assays of sections of human AD brain and of mouse brain. Curcumin also reduced levels of miR-155, a micro-RNA reported to drive a neurodegenerative microglial phenotype. In conditions without amyloid (human microglial cells in vitro, aged wild-type mice), Curc-lo similarly decreased CD33 and increased TREM2. Like curcumin, anti-Aß antibody (also reported to engage the Syk pathway, increase CD68, and decrease amyloid burden in human and mouse brain) increased TREM2 in APPsw mice and decreased amyloid in human AD sections ex vivo. We conclude that curcumin is an immunomodulatory treatment capable of emulating anti-Aß vaccine in stimulating phagocytic clearance of amyloid by reducing CD33 and increasing TREM2 and TyroBP, while restoring neuroinflammatory networks implicated in neurodegenerative diseases.

Influences of miR-320a on proliferation and apoptosis of fibroblast-like synoviocytes in rheumatoid arthritis through targeting MAPK-ERK1/2.

OBJECTIVE: To study the expression of micro ribonucleic acid (miR)-320a in synovial tissues of patients with rheumatoid arthritis (RA) and explore the influences of miR-320a on the proliferation and apoptosis of fibroblast-like synoviocytes (FLSs) in RA and its mechanism. PATIENTS AND METHODS: The expression level of miR-320a in synovial tissues of 40 healthy people and 32 RA patients was detected via reverse transcription-polymerase chain reaction (RT-PCR). The FLSs were isolated from RA patients, cultured in vitro and divided into Control group and miR-320a mimic group. The proliferation and apoptosis of FLSs in each group were observed. Finally, the expression level of mitogen-activated protein kinase (MAPK)-extracellular signal-regulated kinase (ERK) 1/2 in each group was detected via Western blotting. RESULTS: The expression level of miR-320a in synovial tissues of RA patients was significantly lower than that in healthy people (p < 0.05). After miR-320a mimic was transfected into FLSs cultured in vitro, EdU staining and flow cytometry analysis were performed. The results revealed that the proportion of EdU-positive cells significantly declined in miR-320a mimic group, the proportion of cells in G0/G1 phase was increased, while the cells in G2/M and S phases were significantly decreased (p < 0.05). Above data indicated that the cell proliferation ability was significantly inhibited. In addition, the results of flow cytometry also showed that the apoptosis rate of FLSs in miR-320a mimic group was significantly higher than that in Control group (p < 0.05). The results of Western blotting manifested that the Bcl-2 associated X protein (Bax)/Bcl-2 ratio in miR-320a mimic group was also obviously increased (p < 0.05). According to further studies, the phosphorylation level of ERK1/2 in miR-320a mimic group was remarkably inhibited (p < 0.05). CONCLUSIONS: The expression level of miR-320a significantly declined in synovial tissues of RA patients. MiR-320a attenuated proliferation and promoted apoptosis of FLSs through inhibiting the activation of the MAPK-ERK1/2 signaling pathway.