bluuebunny/crossref_metadata_2025 huggingface dataset:
Exploration of the Quorum-Quenching Mechanism in Pseudomonas nitroreducens W-7 and Its Potential to Attenuate the Virulence of Dickeya zeae EC1
Authors: Zhang Wenping , Fan Xinghui , Li Jiayi , Ye Tian , Mishra Sandhya , Zhang Lianhui , Chen Shaohua
Published: August 2021

Quorum quenching (QQ) is a novel, promising strategy that opens up a new perspective for controlling quorum-sensing (QS)-mediated bacterial pathogens. QQ is performed by interfering with population-sensing systems, such as by the inhibition of signal synthesis, catalysis of degrading enzymes, and modification of signals. In many Gram-negative pathogenic bacteria, a class of chemically conserved signaling molecules named N-acyl homoserine lactones (AHLs) have been widely studied. AHLs are involved in the modulation of virulence factors in various bacterial pathogens including Dickeya zeae. Dickeya zeae is the causal agent of plant-rot disease of bananas, rice, maize, potatoes, etc., causing enormous economic losses of crops. In this study, a highly efficient AHL-degrading bacterial strain W-7 was isolated from activated-sludge samples and identified as Pseudomonas nitroreducens. Strain W-7 revealed a superior ability to degrade N-(3-oxododecanoyl)-l-homoserine lactone (OdDHL) and completely degraded 0.2 mmol/L of OdDHL within 48 h. Gas chromatography-mass spectrometry (GC-MS) identified N-cyclohexyl-propanamide as the main intermediate metabolite during AHL biodegradation. A metabolic pathway for AHL in strain W-7 was proposed based on the chemical structure of AHL and intermediate products. In addition to the degradation of OdDHL, this strain was also found to be capable of degrading a wide range of AHLs including N-(3-oxohexanoyl)-l-homoserine lactone (OHHL), N-(3-oxooctanoyl)-l-homoserine lactone (OOHL), and N-hexanoyl-l-homoserine lactone (HHL). Moreover, the application of strain W-7 as a biocontrol agent could substantially attenuate the soft rot caused by D. zeae EC1 to suppress tissue maceration in various host plants. Similarly, the application of crude enzymes of strain W-7 significantly reduced the disease incidence and severity in host plants. These original findings unveil the biochemical aspects of a highly efficient AHL-degrading bacterial isolate and provide useful agents that exhibit great potential for the control of infectious diseases caused by AHL-dependent bacterial pathogens.
Legionella feeleii: Ubiquitous Pathogen in the Environment and Causative Agent of Pneumonia
Authors: Vaccaro Lucianna , Gomes Thiago Santos , Izquierdo Fernando , Magnet Angela , Llorens Berzosa Sergio , Ollero Dolores , Salso Santiago , Alhambra Almudena , Gómez Carmen , López Cano María , Pelaz Carmen , Bellido Samaniego Beatriz , del Aguila Carmen , Fenoy Soledad , Hurtado-Marcos Carolina
Published: August 2021

L. feeleii is one of the most frequent Legionella species isolated from natural pools of the central region of Spain. This study aimed to evaluate its ecology and to identify this Legionella species as a respiratory pathogen. A PCR assay for detecting the L. feeleii mip gene was developed to identify it in clinical and environmental samples. Culture and PCR were performed in environmental samples from four drinking water treatment plants (DWTPs). Free L. feeleii was only detected in raw water samples (3.4%), while L. feeleii as an Acanthamoeba endosymbiont was found in 30.7% of raw water, 11.5% of decanter biofilm, and 32% of finished water samples. Therefore, Acanthamoeba spp. plays an essential role in the multiplication, persistence, and spread of Legionella species in the environment. The first case of Legionnaires’ disease caused by L. feeleii in Spain is described in this study. The case was diagnosed in an older woman through PCR and sequencing from urine and sputum samples. A respiratory infection could be linked with health care procedures, and the patient presented several risk factors (age, insulin-dependent diabetes, and heart disease). The detection of non-L. pneumophila, such as L. feeleii, is a factor that must be considered when establishing or reviewing measures for the control and prevention of legionellosis.
The Role of the Membrane-Associated Domain of the Export Apparatus Protein, EscV (SctV), in the Activity of the Type III Secretion System
Authors: Mitrović Boško , Lezerovich Shir , Sal-Man Neta
Published: August 2021

Diarrheal diseases remain a major public health concern worldwide. Many of the causative bacterial pathogens that cause these diseases have a specialized protein complex, the type III secretion system (T3SS), which delivers effector proteins directly into host cells. These effectors manipulate host cell processes for the benefit of the infecting bacteria. The T3SS structure resembles a syringe anchored within the bacterial membrane, projecting toward the host cell membrane. The entry port of the T3SS substrates, called the export apparatus, is formed by five integral membrane proteins. Among the export apparatus proteins, EscV is the largest, and as it forms a nonamer, it constitutes the largest portion of the export apparatus complex. While there are considerable data on the soluble cytoplasmic domain of EscV, our knowledge of its membrane-associated section and its transmembrane domains (TMDs) is still very limited. In this study, using an isolated genetic reporter system, we found that TMD5 and TMD6 of EscV mediate strong self-oligomerization. Substituting these TMDs within the full-length protein with a random hydrophobic sequence resulted in a complete loss of function of the T3SS, further suggesting that the EscV TMD5 and TMD6 sequences have a functional role in addition to their structural role as membrane anchors. As we observed only mild reduction in the ability of the TMD-exchanged variants to integrate into the full or intermediate T3SS complexes, we concluded that EscV TMD5 and TMD6 are not crucial for the global assembly or stability of the T3SS complex but are rather involved in promoting the necessary TMD–TMD interactions within the complex and the overall TMD orientation to allow channel opening for the entry of T3SS substrates.
Multi-Racial Normative Data for Lobar and Subcortical Brain Volumes in Old Age: Korean and Caucasian Norms May Be Incompatible With Each Other†
Authors: Choi Yu Yong , Lee Jang Jae , Choi Kyu Yeong , Choi Uk-Su , Seo Eun Hyun , Choo IL Han , Kim Hoowon , Song Min-Kyung , Choi Seong-Min , Cho Soo Hyun , Choe Youngshik , Kim Byeong C. , Lee Kun Ho
Published: August 2021

Brain aging is becoming an increasingly important topic, and the norms of brain structures are essential for diagnosing neurodegenerative diseases. However, previous studies of the aging brain have mostly focused on Caucasians, not East Asians. The aim of this paper was to examine ethnic differences in the aging process of brain structures or to determine to what extent ethnicity affects the normative values of lobar and subcortical volumes in clinically normal elderly and the diagnosis in multi-racial patients with Alzheimer's disease (AD). Lobar and subcortical volumes were measured using FreeSurfer from MRI data of 1,686 normal Koreans (age range 59–89) and 851 Caucasian, non-Hispanic subjects in the ADNI and OASIS datasets. The regression models were designed to predict brain volumes, including ethnicity, age, sex, intracranial volume (ICV), magnetic field strength (MFS), and MRI scanner manufacturers as independent variables. Ethnicity had a significant effect for all lobar (|β| > 0.20, p < 0.001) and subcortical regions (|β| > 0.08, p < 0.001) except left pallidus and bilateral ventricles. To demonstrate the validity of the z-score for AD diagnosis, 420 patients and 420 normal controls were selected evenly from the Korean and Caucasian datasets. The four validation groups divided by race and diagnosis were matched on age and sex using a propensity score matching. We analyzed whether and to what extent the ethnicity adjustment improved the diagnostic power of the logistic regression model that was built using the only z-scores of six regions: bilateral temporal cortices, hippocampi, and amygdalae. The performance of the classifier after ethnicity adjustment was significantly improved compared with the classifier before ethnicity adjustment (ΔAUC = 0.10, D = 7.80, p < 0.001; AUC comparison test using bootstrap). Korean AD dementia patients may not be classified by Caucasian norms of brain volumes because the brain regions vulnerable to AD dementia are bigger in normal Korean elderly peoples. Therefore, ethnicity is an essential factor in establishing normative data for regional volumes in brain aging and applying it to the diagnosis of neurodegenerative diseases.
Serial Recall Order and Semantic Features of Category Fluency Words to Study Semantic Memory in Normal Ageing
Authors: De Marco Matteo , Blackburn Daniel J. , Venneri Annalena
Published: August 2021

Background: Category Fluency Test (CFT) is a common measure of semantic memory (SM). Test performance, however, is also influenced by other cognitive functions. We here propose a scoring procedure that quantifies the correlation between the serial recall order (SRO) of words retrieved during the CFT and a number of linguistic features, to obtain purer SM measures. To put this methodology to the test, we addressed a proof-of-concept hypothesis whereby, in alignment with the literature, older adults would show better SM.

Methods: Ninety participants (45 aged 18–21 years; 45 aged 70–81 years) with normal neurological and cognitive functioning completed a 1-min CFT. SRO was scored as an ordinal variable incrementing by one unit for each valid entry. Each word was also scored for 16 additional linguistic features. Participant-specific normalised correlation coefficients were calculated between SRO and each feature and were analysed with group comparisons and graph theory.

Results: Younger adults showed more negative correlations between SRO and “valence” (a feature of words pleasantness). This was driven by the first five words generated. When analysed with graph theory, SRO had significantly higher degree and lower betweenness centrality among older adults.

Conclusion: In older adults, SM relies significantly less on pleasantness of entries typically retrieved without semantic control. Moreover, graph-theory metrics indicated better optimised links between SRO and linguistic features in this group. These findings are aligned with the principle whereby SM processes tend to solidify with ageing. Although additional work is needed in support of an SRO-based item-level scoring procedure of CFT performance, these initial findings suggest that this methodology could be of help in characterising SM in a purer form.
Habituation After Deep Brain Stimulation in Tremor Syndromes: Prevalence, Risk Factors and Long-Term Outcomes
Authors: Peters James , Tisch Stephen
Published: August 2021

Deep brain stimulation (DBS) of the thalamus is an effective treatment for medically refractory essential, dystonic and Parkinson's tremor. It may also provide benefit in less common tremor syndromes including, post-traumatic, cerebellar, Holmes, neuropathic and orthostatic tremor. The long-term benefit of DBS in essential and dystonic tremor (ET/DT) often wanes over time, a phenomena referred to as stimulation “tolerance” or “habituation”. While habituation is generally accepted to exist, it remains controversial. Attempts to quantify habituation have revealed conflicting reports. Placebo effects, loss of micro-lesional effect, disease related progression, suboptimal stimulation and stimulation related side-effects may all contribute to the loss of sustained long-term therapeutic effect. Habituation often presents as substantial loss of initial DBS benefit occurring as early as a few months after initial stimulation; a complex and feared issue when faced in the setting of optimal electrode placement. Simply increasing stimulation current tends only to propagate tremor severity and induce stimulation related side effects. The report by Paschen and colleagues of worsening tremor scores in the “On” vs. “Off” stimulation state over time, even after accounting for “rebound” tremor, supports the concept of habituation. However, these findings have not been consistent across all studies. Chronic high intensity stimulation has been hypothesized to induce detrimental plastic effects on tremor networks, with some lines of evidence that DT and ET may be more susceptible than Parkinson's tremor to habituation. However, Tsuboi and colleague's recent longitudinal follow-up in dystonic and “pure” essential tremor suggests otherwise. Alternatively, post-mortem findings support a biological adaption to stimulation. The prevalence and etiology of habituation is still not fully understood and management remains difficult. A recent study reported that alternating thalamic stimulation parameters at weekly intervals provided improved stability of tremor control consistent with reduced habituation. In this article the available evidence for habituation after DBS for tremor syndromes is reviewed; including its prevalence, time-course, possible mechanisms; along with expected long-term outcomes for tremor and factors that may assist in predicting, preventing and managing habituation.
Adherence to Low Carbohydrate Diet in Relation to Chronic Obstructive Pulmonary Disease
Authors: Malmir Hanieh , Onvani Shokouh , Ardestani Mohammad Emami , Feizi Awat , Azadbakht Leila , Esmaillzadeh Ahmad
Published: August 2021

Purpose: Data on the link between adherence to low-carbohydrate diet (LCD) and odds of chronic obstructive pulmonary disease (COPD) are scarce. The current study aimed to investigate the relation between adherence to LCD and COPD in Iranian adults.

Methods: In this hospital-based case-control study, we enrolled 84 newly-diagnosed COPD patients and 252 age and sex matched healthy controls in Alzahra University Hospital, Isfahan, Iran. COPD was defined based on findings of spirometry test (forced expiratory volume in 1 s (FEV1)/forced vital capacity (FVC) < 70% or FEV1 < 80%). Dietary intakes of study participants were assessed using the validated Block-format 168-item FFQ. Data on potential confounders were also collected through the use of a pre-tested questionnaire.

Results: Mean age of cases and controls were 57.7 and 55.07 years, respectively. Adherence to LCD was inversely associated with odds of COPD (0.35; 95% CI: 0.16-0.75). This inverse association did not alter after controlling for age, sex, and energy intake (0.42; 95% CI: 0.19-0.93). Adjustments for other potential confounders, including dietary intakes, smoking, and educational status, did not affect these findings; such that those in the highest quintile of LCD score were 64% less likely to have COPD than those in the lowest quintile (OR: 0.36; 95% CI: 0.13-0.99).

Conclusion: We found an inverse association between adherence to LCD and odds of COPD. The association remained statistically significant even after taking other potential confounders, including socioeconomic characteristic and dietary intakes into account.
Performances of Whole Tumor Texture Analysis Based on MRI: Predicting Preoperative T Stage of Rectal Carcinomas
Authors: You Jia , Yin Jiandong
Published: August 2021

Objective

To determine whether there is a correlation between texture features extracted from high-resolution T2-weighted imaging (HR-T2WI) or apparent diffusion coefficient (ADC) maps and the preoperative T stage (stages T1–2 versus T3–4) in rectal carcinomas.

Materials and Methods

One hundred and fifty four patients with rectal carcinomas who underwent preoperative HR-T2WI and diffusion-weighted imaging were enrolled. Patients were divided into training (n = 89) and validation (n = 65) cohorts. 3D Slicer was used to segment the entire volume of interest for whole tumors based on HR-T2WI and ADC maps. The least absolute shrinkage and selection operator (LASSO) was performed to select feature. The significantly difference was tested by the independent sample t-test and Mann-Whitney U test. The support vector machine (SVM) model was used to develop classification models. The correlation between features and T stage was assessed by Spearman’s correlation analysis. Multivariate logistic regression analysis was performed to identify independent predictors of tumor invasion. The performance of classifiers was evaluated by the receiver operating characteristic (ROC) curves.

Results

The wavelet HHH NGTDM strength (RS = -0.364, P < 0.001) from HR-T2WI was an independent predictor of stage T3–4 tumors. The shape maximum 2D diameter column (RS = 0.431, P < 0.001), log σ = 5.0 mm 3D first-order maximum (RS = 0.276, P = 0.009), and log σ = 5.0 mm 3D first-order interquartile range (RS = -0.229, P = 0.032) from ADC maps were independent predictors. In training cohorts, the classification models from HR-T2WI, ADC maps and the combination of two achieved the area under the ROC curves (AUCs) of 0.877, 0.902 and 0.941, with the accuracy of 79.78%, 89.86% and 89.89%, respectively. In validation cohorts, the three models achieved AUCs of 0.845, 0.881 and 0.910, with the accuracy of 78.46%, 83.08% and 87.69%, respectively.

Conclusions

Texture analysis based on ADC maps shows more potential than HR-T2WI in identifying preoperative T stage in rectal carcinomas. The combined application of HR-T2WI and ADC maps may help to improve the accuracy of preoperative diagnosis of rectal cancer invasion.
A Novel Prognostic Marker for Primary CNS Lymphoma: Lactate Dehydrogenase-to-Lymphocyte Ratio Improves Stratification of Patients Within the Low and Intermediate MSKCC Risk Groups
Authors: Gao Yuting , Wei Li , Kim Seok Jin , Wang Liang , He Yingzhi , Zheng Yanfang , Bertero Luca , Pellerino Alessia , Cassoni Paola , Tamagnone Luca , Theresa Prochazka Katharina , Deutsch Alexander , Zhan Huien , Lai Jing , Wang Yao , You Hua
Published: August 2021

Background

Primary central nervous system lymphoma (PCNSL) is a highly aggressive and rare extranodal non-Hodgkin lymphoma (NHL). The MSKCC and the IELSG scores represent the most widely used prognostic models, but many changes have occurred in therapeutic protocols since their development. Moreover, many PCNSL patients cannot be classified using the IELSG score. We thus aimed to create a novel, effective and feasible prognostic model for PCNSL.

Methods

We included 248 PCNSL patients diagnosed with PCNSL. Our primary endpoint was the overall survival (OS) and we used the receiver operating characteristic (ROC) analysis to determine the optimal prognostic cut-off value for LLR (lactate dehydrogenase-to-lymphocyte ratio), neutrophil-to-lymphocyte ratio (NLR) and derived neutrophil-to-lymphocyte ratio (dNLR). Variable associated with OS were evaluated by univariate and multivariate analyses. 124 out of 248 patients were randomly selected as the internal validation cohort.

Results

By univariate analysis, an age >60 years, Eastern Cooperative Oncology Group performance status (ECOG PS) >1, treatment with radiotherapy alone, high-risk groups of Memorial Sloan Kettering Cancer Center (MSKCC) score, NLR >4.74, dNLR >3.29, and LLR >166.8 were significantly associated with a worse OS. By multivariate analysis, the MSKCC score and LLR were confirmed as independent prognostic parameters for poorer OS. OS, however, was not significantly different between low- and intermediate-risk groups according to the MSKCC score, while LLR proved to be prognostically relevant and was thus used to develop a novel, effective three-tier PCNSL scoring system. Of 124 patients, 84 patients with survival data and LLR data were successfully validated by newly established PCNSL LLR scoring system.

Conclusions

In the present study, we demonstrate that a high LLR represents an independent unfavorable prognostic parameter in PCNSL patients which can be integrated into an effective prognostic model.
Acetate Promotes a Differential Energy Metabolic Response in Human HCT 116 and COLO 205 Colon Cancer Cells Impacting Cancer Cell Growth and Invasiveness
Authors: Rodríguez-Enríquez Sara , Robledo-Cadena Diana Xochiquetzal , Gallardo-Pérez Juan Carlos , Pacheco-Velázquez Silvia Cecilia , Vázquez Citlali , Saavedra Emma , Vargas-Navarro Jorge Luis , Blanco-Carpintero Betsy Alejandra , Marín-Hernández Álvaro , Jasso-Chávez Ricardo , Encalada Rusely , Ruiz-Godoy Luz , Aguilar-Ponce José Luis , Moreno-Sánchez Rafael
Published: August 2021

Under dysbiosis, a gut metabolic disorder, short-chain carboxylic acids (SCCAs) are secreted to the lumen, affecting colorectal cancer (CRC) development. Butyrate and propionate act as CRC growth inhibitors, but they might also serve as carbon source. In turn, the roles of acetate as metabolic fuel and protein acetylation promoter have not been clearly elucidated. To assess whether acetate favors CRC growth through active mitochondrial catabolism, a systematic study evaluating acetate thiokinase (AcK), energy metabolism, cell proliferation, and invasiveness was performed in two CRC cell lines incubated with physiological SCCAs concentrations. In COLO 205, acetate (+glucose) increased the cell density (50%), mitochondrial protein content (3–10 times), 2-OGDH acetylation, and oxidative phosphorylation (OxPhos) flux (36%), whereas glycolysis remained unchanged vs. glucose-cultured cells; the acetate-induced OxPhos activation correlated with a high AcK activity, content, and acetylation (1.5–6-fold). In contrast, acetate showed no effect on HCT116 cell growth, OxPhos, AcK activity, protein content, and acetylation. However, a substantial increment in the HIF-1α content, HIF-1α-glycolytic protein targets (1–2.3 times), and glycolytic flux (64%) was observed. Butyrate and propionate decreased the growth of both CRC cells by impairing OxPhos flux through mitophagy and mitochondrial fragmentation activation. It is described, for the first time, the role of acetate as metabolic fuel for ATP supply in CRC COLO 205 cells to sustain proliferation, aside from its well-known role as protein epigenetic regulator. The level of AcK determined in COLO 205 cells was similar to that found in human CRC biopsies, showing its potential role as metabolic marker.
Urban–Rural Differences in Clinical Characteristics of Prostate Cancer at Initial Diagnosis: A Single-Center Observational Study in Anhui Province, China
Authors: Song Qi Long , Qian Yinfeng , Min Xuhong , Wang Xiao , Wu Jing , Li Xiaohu , Yu Yongqiang
Published: August 2021

Background

People residing in rural areas have higher prostate cancer (PCa) mortality to incidence ratio (M/I) and worse prognosis than those in urban areas of China. Clinical characteristics at initial diagnosis are significantly associated with biochemical recurrence, overall survival, and PCa disease-free survival.

Objective

This study aimed at investigating the clinical characteristics at initial diagnosis of urban and rural PCa patients and to establish a logistic regression model for identifying independent predictors for high-grade PCa.

Materials and Methods

Clinical characteristics for PCa patients were collected from the largest prostate biopsy center in Anhui province, China, from December 2015 to March 2019. First, urban–rural disparities in clinical characteristics were evaluated at initial diagnosis. Second, based on pathological findings, we classified all participants into the benign+ low/intermediate-grade PCa or high-grade PCa groups. Univariate and multivariate logistic regression analyses were performed to identify independent factors for predicting high-grade PCa, while a nomogram for predicting high-grade PCa was generated based on all independent factors. The model was evaluated using area under receiver-operating characteristic (ROC) curve as well as calibration curve analyses and compared to a model without the place of residence factor of individuals.

Results

Statistically significant differences were observed between urban and rural PCa patients with regard to tPSA, PSA density (PSAD), and Gleason score (GS) (p < 0.05). Logistic regression analysis revealed that tPSA [OR = 1.060, 95% confidence interval (CI): 1.024, 1.098], PSAD (OR = 14.678, 95%CI: 4.137, 52.071), place of residence of individuals (OR = 5.900, 95%CI: 1.068, 32.601), and prostate imaging reporting and data system version 2 (PI-RADS v2) (OR = 4.360, 95%CI: 1.953, 9.733) were independent predictive factors for high-grade PCa. The area under the curve (AUC) of the nomogram was greater than that of the model without the place of residence of individuals. The calibration curve of the nomogram indicated that the prediction curve was basically fitted to the standard curve, suggesting that the prediction model had a better calibration ability.

Conclusions

Compared to urban PCa patients, rural PCa patients presented elevated tPSA, PSAD levels, and higher pathological grades. The place of residence of the individuals was an independent predictor for high-grade PCa in Anhui Province, China. Therefore, appropriate strategies, such as narrowing urban-rural gaps in access to health care and increasing awareness on the importance of early detection should be implemented to reduce PCa mortality rates.
A Parallel Comparison Method of Early Gastric Cancer: The Light Transmission-Assisted Pathological Examination of Specimens of Endoscopic Submucosal Dissection
Authors: Chen Guangyong , Xu Rui , Yue Bing , Jia Mei , Li Peng , Ji Ming , Zhang Shutian
Published: August 2021

Objective

It is always challenging to diagnose and characterize early gastric cancer surrounded by non-cancerous mucosa, including the malignant diagnosis and extent and depth of the lesions. Therefore, we developed a light transmission-assisted pathological examination to diagnose and characterize early gastric cancer. Here, we performed a parallel comparison between the light transmission-assisted pathological examination under endoscopy and the histological examination for the diagnosis of early gastric cancer.

Methods

First, the endoscopic submucosal dissection (ESD) specimen was first placed on the surface of the light-emitting diode lamp to observe the mucosal surface structure and blood vessels. Second, the sliced and embedded tissue strips were cut into 3-µm sections for hematoxylin and eosin staining. Third, the histopathology of each section was projected onto a macroscopic image. Finally, the macroscopic and microscopic changes in the ESD specimens observed under endoscopy were compared. Seventy cases of early gastric adenocarcinoma were diagnosed and characterized using this new method.

Results

Using the conventional pathological method, the demarcation line of the lesions was seen in 40 of 70 (57.1%) cases. Furthermore, no surface structure or microvascular changes were observed in any of the cases. Based on the light transmission-assisted pathological examination, 58 of 70 (82.9%) cases presented clear edges of neoplastic and non-neoplastic epithelia, with a classifiable surface structure (88.6%) and microvascular type (78.8%).

Conclusions

This pilot method provided a practical bridge between endoscopic and pathological examinations. Compared to the histological examination, the light transmission-assisted pathological examination was an easier and more precise way to match the in vivo endoscopic observation and in vitro pathological examination.
A Novel Prognostic Model Based on Autophagy-Related Long Non-Coding RNAs for Clear Cell Renal Cell Carcinoma
Authors: Li Xinyuan , Yu Haitao , Wei Zongjie , Gou Xin , Liang Simin , Liu Fu
Published: August 2021

Background

Renal cell carcinoma (RCC) is one of the most common malignant tumors of the urinary system, of which the clear cell renal cell carcinoma (ccRCC) accounts for the most subtypes. The increasing discoveries of abundant autophagy-related long non-coding RNAs (ARLNRs) lead to a resurgent interest in evaluating their potential on prognosis prediction. Based on a large number of ccRCC gene samples from TCGA and clinics, ARLNRs analysis will provide a novel perspective into this field.

Methods

We calculated the autophagy scores of each sample according to the expression levels of autophagy-related genes (ARGs) and screened the survival-related ARLNRs (sARLNRs) of ccRCC patients by Cox regression analysis. The high-risk group and the low-risk group were distinguished by the median score of the autophagy-related risk score (ARRS) model. The functional annotations were detected by gene set enrichment analysis (GSEA) and principal component analysis (PCA). The expression levels of two kinds of sARLNRs in the renal tumor and adjacent normal tissues and cell lines were verified.

Results

There were 146 ARLNRs selected by Pearson analysis. A total of 30 sARLNRs were remarkably correlated with the clinical outcomes of ccRCC patients. Eleven sARLNRs (AC002553.1, AC092611.2, AL360181.2, AP002807.1, AC098484.1, AL513218.1, AC008735.2, MHENCR, AC020907.4, AC011462.4, and AC008870.2) with the highest prognosis value were recruited to establish the ARRS in which the overall survival (OS) in the high-risk group was shorter than that in the low-risk group. ARRS could be treated as an independent prognostic factor and has significant correlations with OS. The distributions of autophagy genes were different between the high-risk group and the low-risk group. In addition, we also found that the expression levels of AC098484.1 in ccRCC cell lines and tumor tissues were lower than those in HK-2 and adjacent normal tissues, but AL513218.1 showed the inverse level. Furthermore, the AC098484.1 expressed decreasingly with the more advanced T-stages, but AL513218.1 gradually increased.

Conclusion

Our study identified and verified some sARLNRs with clinical significances and revealed their potential values on predicting prognoses of ccRCC patients, which may provide a novel perspective for autophagy-related research and clinical decisions.
Role of Connexin 43 in an Inflammatory Model for TMJ Hyperalgesia
Authors: Ahmed Fabeeha , Rahman Md. , Thompson Randall , Bereiter David A.
Published: August 2021

Temporomandibular joint disorders (TMD) consist of a heterogeneous group of conditions that present with pain in the temporomandibular joint (TMJ) region and muscles of mastication. This project assessed the role of connexin 43 (Cx43), a gap junction protein, in the trigeminal ganglion (TG) in an animal model for persistent inflammatory TMJ hyperalgesia. Experiments were performed in male and female rats to determine if sex differences influence the expression and/or function of Cx43 in persistent TMJ hyperalgesia. Intra-TMJ injection of Complete Freund's Adjuvant (CFA) caused a significant increase in Cx43 expression in the TG at 4 days and 10 days post-injection in ovariectomized (OvX) female rats and OvX females treated with estradiol (OvXE), while TG samples in males revealed only marginal increases. Intra-TG injection of interference RNA for Cx43 (siRNA Cx43) 3 days prior to recording, markedly reduced TMJ-evoked masseter muscle electromyographic (MMemg) activity in all CFA-inflamed rats, while activity in sham animals was not affected. Western blot analysis revealed that at 3 days after intra-TG injection of siRNA Cx43 protein levels for Cx43 were significantly reduced in TG samples of all CFA-inflamed rats. Intra-TG injection of the mimetic peptide GAP19, which inhibits Cx43 hemichannel formation, greatly reduced TMJ-evoked MMemg activity in all CFA-inflamed groups, while activity in sham groups was not affected. These results revealed that TMJ inflammation caused a persistent increase in Cx43 protein in the TG in a sex-dependent manner. However, intra-TG blockade of Cx43 by siRNA or by GAP19 significantly reduced TMJ-evoked MMemg activity in both males and females following TMJ inflammation. These results indicated that Cx43 was necessary for enhanced jaw muscle activity after TMJ inflammation in males and females, a result that could not be predicted on the basis of TG expression of Cx43 alone.
Examining the Acute Effects of Classroom-Based Physical Activity Breaks on Executive Functioning in 11- to 14-Year-Old Children: Single and Additive Moderation Effects of Physical Fitness
Authors: Graham Jeffrey D. , Bremer Emily , Fenesi Barbara , Cairney John
Published: August 2021

Objective: Research supports the efficacy of acute, classroom-based, physical activity breaks on executive functioning in children. However, research pertaining to the effect of physical fitness on the acute physical activity—executive functioning relationship remains limited. The primary purpose of this study was to investigate the acute effects of classroom-based, teacher-delivered, physical activity breaks on executive functioning in 11–14-year-old children. We also investigated the potential moderating effects of both aerobic and musculoskeletal fitness on the acute physical activity—executive functioning relationship.

Method: Participants (N = 116) completed pre- and post-test assessments of executive functioning (i.e., inhibition, switching, and updating) separated by a classroom-based physical activity break or sedentary classroom work. We manipulated the dose (i.e., length) and type of physical activity breaks. With regards to dose, participants in the experimental conditions engaged in 5-, 10-, or 20-min of physical activity whereas controls completed sedentary classroom math work at their desk. With regards to type, one experimental condition completed traditional physical activity breaks whereas the other experimental condition completed academic physical activity breaks (i.e., performed mental math and physical activity). Participants' mood, motivation, and self-efficacy were also assessed following the experimental manipulations.

Results: Overall, executive function scores improved across each assessment following the physical activity breaks when compared to sedentary classroom work regardless of dose and type. Participants also reported more positive mood states, higher motivation to complete the executive function tests, and higher self-efficacy to perform the executive functions tests following the physical activity breaks. Single moderation analyses showed that low-moderate levels of aerobic fitness moderated the acute physical activity—executive functioning relationship. Additive moderation analysis showed, collectively, that both aerobic and musculoskeletal fitness moderated the acute physical activity—executive functioning relationship.

Conclusion: Findings from the present study provide evidence for the acute effects of short (i.e., 5–20 min) classroom-based physical activity breaks on executive functioning and psychological states in children. Results also suggest levels of both aerobic and musculoskeletal fitness moderate these effects, however future research is needed to further elucidate this complex relationship.
The Association Between Variability in Electrolytes and the In-Hospital Mortality in Critically Ill Children in Pediatric Intensive Care Units
Authors: Lin Jilei , Zhang Yin , Chen Meng , Dai Jihong , Song Anchao , Chen Jianchuan , Tao Xingping
Published: August 2021

Objective: This study aimed to explore the association between the variability in electrolytes and the in-hospital mortality in critically ill children admitted into intensive care units (ICUs).

Design: This is a retrospective case–control study.

Setting and Participants: Total of 11,245 children have been admitted to ICUs of Children's Hospital of Zhejiang University from 2010 to 2018.

Methods: The coefficient of variation (CV), standard deviation (SD), and variability independent of the mean (VIM) were calculated as variability indices. High variability was defined as having values in the highest quartile for each parameter. Age, sex, diagnoses of disease, and surgical treatment were adjusted in the multivariable-adjusted logistic regression model.

Results: A total of 11,245 children were included, and 660 patients died in the hospital. The median (P25, P75) potassium, sodium, and chloride of all patients were 3.8 (3.58, 4.09), 136.83 (135.11, 138.60), and 108.67 (105.71, 111.17), respectively. U-shaped relationships between the mean, lowest, and highest levels of potassium, sodium, and chloride and the in-hospital mortality were observed. The lowest mortality was noted when serum potassium, sodium, and chloride were between ~3.5 and 5.0, 135 and 145, and 105 and 115 mmol/l, respectively. The areas under the curve (AUCs) of three indices of variability in electrolytes were larger than those of the mean and lowest levels of electrolytes in predicting the in-hospital mortality. In the multivariable-adjusted model, the odds ratios and 95% confidence interval (CI) of the in-hospital mortality were 3.14 (2.44–4.04) for one parameter, 5.85 (4.54–7.53) for two parameters, and 10.32 (7.81–13.64) for three parameters compared with subjects having no parameters of high variability measured as the CV. The results were consistent when the variability was determined using the SD and VIM (all P for trend <0.001). Consistent results were noted in various subgroup analyses.

Conclusions: This study showed that individuals with higher variability of each parameter were related with higher risk of in-hospital mortality. There was a linear association between the number of high variability parameters and the in-hospital mortality. The variability of electrolytes might be a good predictor for in-hospital mortality of children in ICUs.
Whole Exome Sequencing Confirms Molecular Diagnostics of Three Pakhtun Families With Autosomal Recessive Epidermolysis Bullosa
Authors: Fozia Fozia , Nazli Rubina , Bibi Nousheen , Khan Sher Alam , Muhammad Noor , Shakeeb Nafila , Khan Saadullah , Jelani Musharraf , Wasif Naveed
Published: August 2021

Epidermolysis bullosa (EB) is a genetic skin disorder that shows heterogeneous clinical fragility. The patients develop skin blisters congenitally or in the early years of life at the dermo-epithelial junctions, including erosions, hyperkeratosis over the palms and soles. The other associated features are hypotrichosis on the scalp, absent or dystrophic nails, and dental anomalies. Molecular diagnosis through whole-exome sequencing (WES) has become one of the successful tool in clinical setups. In this study, three Pakhtun families from the Khyber Pakhtunkhwa province of Pakistan were ascertained. WES analysis of a proband in each family revealed two novel variants (COL17A1: NM_000494.4: c.4041T>G: p.Y1347* and PLEC: NM_201380.3: c.1283_1285delGCT: p.L426del) and one previously known COL17A1: NM_000494.4:c.3067C>T: p.Q1023*) variant in homozygous forms. Sanger sequencing of the identified variants confirmed that the heterozygous genotypes of the obligate carriers. The identified variants have not only increased the mutation spectrum of the COL17A1 and PLEC but also confirms their vital role in the morphogenesis of skin and its associated appendages. WES can be used as a first-line diagnostic tool in genetic testing and counselling families from Khyber Pakhtunkhwa, Pakistan.
Antitumor Effect of Fluoxetine on Chronic Stress-Promoted Lung Cancer Growth via Suppressing Kynurenine Pathway and Enhancing Cellular Immunity
Authors: Yang Zhen , Li Zhuman , Guo Zhijun , Ren Yu , Zhou Ting , Xiao Zhijun , Duan Jingjing , Han Chuangchuang , Cheng Yuanchi , Xu Feng
Published: August 2021

Background: Chronic stress promotes cancer growth. Antidepressant fluoxetine (FLX) is usually prescribed for cancer patients with comorbid depression. FLX displays inhibition on cancer cell proliferation, however, the in vivo activity has not been investigated.

Methods: We explored the antitumor effect of FLX in subcutaneous transplanted lung cancer cells in a tumor-bearing mouse model. Fifty-six C57BL/6 mice were randomly divided into group A (blank control), group B (tumor-bearing control), group C (tumor-bearing + FLX), group D (CUMS control), group E (CUMS + FLX), group F (tumor-bearing + CUMS), and group G (tumor-bearing + CUMS + FLX). 5-HT, tryptophane (Trp), kynurenine, IFN-γ, TNF-α, IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-17A levels were measured by ELISA. T helper (Th), cytotoxic T (Tc) and regulatory T cells (Tregs) subtype were measured by flow cytometry. The antitumor effects of FLX were evaluated by tumor weight. The expression of kynurenine pathway related genes TDO, IDO1, IDO2, and apoptosis-related genes caspase1, 3, 4, 5, 7, 12 in tumor tissues were measured by western blotting and qRT-PCR. A549 cells were exposed with FLX (15 μmol/L) and its effect on cell proliferation, migration, and clonal formation were detected. Kynurenine pathway and apoptosis related gene expression were also measured.

Results:In vivo, chronic stress promoted tumor growth in C57BL/6 mice. FLX administration not only significantly reversed chronic unpredictable mild stress (CUMS)-induced reduction of 5-HT and Trp, increment of kynurenine, but increased CD4+ Th and CD8+ Tc cells, and reduced CD25+ FOXP3+ Tregs. FLX promoted Th to differentiate into Th1 cells and increased IL-2 and IFN-γ, meanwhile inhibited Th differentiate into Th2 and Th17 cells and decreased the concentrations of IL-4, IL-6, IL-10, and IL-17A. Chronic stress obviously up-regulated IDO1 and IDO2 expression, down-regulated caspase 4, 7, and 12 expression, meanwhile FLX administration reversed this regulation. However, there was no significant change in TDO, caspase 1, 3, 5. Similarly, in vitro, FLX administration significantly inhibited the proliferation, migration, and clonal formation of A549 cells and induced cell apoptosis. FLX administration down-regulated the expression of IDO1, IDO2, and up-regulated caspase 4, 5, and 7.

Conclusion: Fluoxetine administration could inhibit tumor growth. The inhibition might be via suppressing kynurenine pathway and enhancing cellular immunity.
A Polyphenol-Rich Extract of Olive Mill Wastewater Enhances Cancer Chemotherapy Effects, While Mitigating Cardiac Toxicity
Authors: Albini Adriana , Festa Marco M. G. , Ring Nadja , Baci Denisa , Rehman Michael , Finzi Giovanna , Sessa Fausto , Zacchigna Serena , Bruno Antonino , Noonan Douglas M.
Published: August 2021

Cardiovascular toxicity remains one of the most adverse side effects in cancer patients receiving chemotherapy. Extra-virgin olive oil (EVOO) is rich in cancer preventive polyphenols endowed with anti-inflammatory, anti-oxidant activities which could exert protective effects on heart cells. One very interesting derivative of EVOO preparation is represented by purified extracts from olive mill waste waters (OMWW) rich in polyphenols. Here, we have investigated the anti-cancer activity of a OMWW preparation, named A009, when combined with chemotherapeutics, as well as its potential cardioprotective activities. Mice bearing prostate cancer (PCa) xenografts were treated with cisplatin, alone or in combination with A009. In an in vivo model, we found synergisms of A009 and cisplatin in reduction of prostate cancer tumor weight. Hearts of mice were analyzed, and the mitochondria were studied by transmission electron microscopy. The hearts of mice co-treated with A009 extracts along with cisplatin had reduced mitochondria damage compared to the those treated with chemotherapy alone, indicating a cardioprotective role. To confirm the in vivo results, tumor cell lines and rat cardiomyocytes were treated with cisplatin in vitro, with and without A009. Another frequently used chemotherapeutic agent 5-fluorouracil (5-FU), was also tested in this assay, observing a similar effect. In vitro, the combination of A009 with cisplatin or 5-FU was effective in decreasing prostate and colon cancer cell growth, while it did not further reduce growth of rat cardiomyocytes also treated with cisplatin or 5-FU. A009 cardioprotective effects towards side effects caused by 5-FU chemotherapy were further investigated, using cardiomyocytes freshly isolated from mice pups. A009 mitigated toxicity of 5-FU on primary cultures of mouse cardiomyocytes. Our study demonstrates that the polyphenol rich purified A009 extracts enhance the effect of chemotherapy in vitro and in vivo, but mitigates chemotherpy adverse effects on heart and on isolated cardiomyocytes. Olive mill waste water extracts could therefore represent a potential candidate for cardiovascular prevention in patients undergoing cancer chemotherapy.
ZnT8 Deficiency Protects From APAP-Induced Acute Liver Injury by Reducing Oxidative Stress Through Upregulating Hepatic Zinc and Metallothioneins
Authors: Su Wen , Feng Mingji , Liu Yuan , Cao Rong , Liu Yiao , Tang Junyao , Pan Ke , Lan Rongfeng , Mao Zhuo
Published: August 2021

Zinc transporter 8 (ZnT8) is an important zinc transporter highly expressed in pancreatic islets. Deficiency of ZnT8 leads to a marked decrease in islet zinc, which is thought to prevent liver diseases associated with oxidative stress. Herein, we aimed to investigate whether loss of islet zinc affects the antioxidant capacity of the liver and acute drug-induced liver injury. To address this question, we treated ZnT8 knockout (KO) or wild-type control mice with 300 mg/ kg acetaminophen (APAP) or phosphate-buffered saline (PBS). Unexpectedly, we found that loss of ZnT8 in mice ameliorated APAP-induced injury and was accompanied by inhibition of c-Jun N-terminal kinase (JNK) activation, reduced hepatocyte death, and decreased serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). An increase in hepatic glutathione (GSH) was observed, corresponding to a decrease in malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) levels. APAP-induced inflammation and glycogen depletion were alleviated. In contrast, no significant changes were observed in cytochrome P450 family 2 subfamily E member 1 (CYP2E1), the main enzyme responsible for drug metabolism. Elevated levels of hepatic zinc and metallothionein (MT) were also observed, which may contribute to the hepatoprotective effect in ZnT8 KO mice. Taken together, these results suggest that ZnT8 deficiency protects the liver from APAP toxicity by attenuating oxidative stress and promoting hepatocyte proliferation. This study provides new insights into the functions of ZnT8 and zinc as key mediators linking pancreatic and hepatic functions.
Utilization of Transcriptome, Small RNA, and Degradome Sequencing to Provide Insights Into Drought Stress and Rewatering Treatment in Medicago ruthenica
Authors: Shi Rui , Jiao Wei , Yun Lan , Zhang Zhiqiang , Zhang Xiujuan , Wang Quanzhen , Li Ying , Mi Fugui
Published: August 2021

Drought is a major limiting factor in foraging grass yield and quality. Medicago ruthenica (M. ruthenica) is a high-quality forage legume with drought resistance, cold tolerance, and strong adaptability. In this study, we integrated transcriptome, small RNA, and degradome sequencing in identifying drought response genes, microRNAs (miRNAs), and key miRNA-target pairs in M. ruthenica under drought and rewatering treatment conditions. A total of 3,905 genes and 50 miRNAs (45 conserved and 5 novel miRNAs) were significantly differentially expressed in three test conditions (CK: control, DS: plants under drought stress, and RW: plants rewatering after drought stress). The degradome sequencing (AllenScore < 4) analysis revealed that 104 miRNAs (11 novel and 93 conserved miRNAs) were identified with 263 target transcripts, forming 296 miRNA-target pairs in three libraries. There were 38 differentially expressed targets from 16 miRNAs in DS vs. CK, 31 from 11 miRNAs in DS vs. RW, and 6 from 3 miRNAs in RW vs. CK; 21, 18, and 3 miRNA-target gene pairs showed reverse expression patterns in DS vs. CK, DS vs. RW, and RW vs. CK comparison groups, respectively. These findings provide valuable information for further functional characterization of genes and miRNAs in response to abiotic stress, in general, and drought stress in M. ruthenica, and potentially contribute to drought resistance breeding of forage in the future.