bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular diagnosis of bovine genital campylobacteriosis using high-resolution melting analysis
Authors: Silva Marta Filipa , Kienesberger Sabine , Pereira Gonçalo , Mateus Luísa , Lopes-da-Costa Luís , Silva Elisabete
Published: September 2022

Bovine Genital Campylobacteriosis (BGC) is a worldwide spread venereal disease of cattle caused by Campylobacter fetus subsp. venerealis (Cfv). Although several real-time PCR assays were developed for Cfv identification, most target mobile genetic elements, which may lead to false-positive diagnosis. In this study, a real-time PCR assay coupled with High-Resolution Melting analysis (HRM) was developed for the identification of Campylobacter fetus subspecies and application in BGC diagnosis. Two HRM assays targeting different single nucleotide polymorphisms were validated using 51 C. fetus strains, including 36 Cfv and 15 C. fetus subsp. fetus (Cff). The specificity was assessed in 50 preputial samples previously tested as negative for C. fetus and in 24 strains from other Campylobacter species. The analytical sensitivity was determined with ten-fold dilutions of Cfv genome copies and in preputial samples spiked with Cfv cells. Both HRM assays accurately identified the 51 C. fetus strains, showing 100% concordance with the previous identification. C. fetus subspecies identification by HRM showed concordant results with the glycine test in 98.0% of the isolates. No amplification was obtained in C. fetus negative preputial samples as well as in strains from other Campylobacter species. The assays were able to detect 102 genome copies of Cfv, while for preputial washing samples the limit of detection was 103 CFU/mL. These novel HRM assays represent a highly specific and sensitive tool for the identification of C. fetus subspecies and show potential for direct use in bull preputial samples for BGC diagnosis.
Signal and regulatory mechanisms involved in spore development of Phytophthora and Peronophythora
Authors: Situ Junjian , Xi Pinggen , Lin Long , Huang Weixiong , Song Yu , Jiang Zide , Kong Guanghui
Published: September 2022

Oomycetes cause hundreds of destructive plant diseases, threatening agricultural production and food security. These fungus-like eukaryotes show multiple sporulation pattern including the production of sporangium, zoospore, chlamydospore and oospore, which are critical for their survival, dispersal and infection on hosts. Recently, genomic and genetic technologies have greatly promoted the study of molecular mechanism of sporulation in the genus Phytophthora and Peronophythora. In this paper, we characterize the types of asexual and sexual spores and review latest progress of these two genera. We summarize the genes encoding G protein, mitogen-activated protein kinase (MAPK) cascade, transcription factors, RNA-binding protein, autophagy-related proteins and so on, which function in the processes of sporangium production and cleavage, zoospore behaviors and oospore formation. Meanwhile, various molecular, chemical and electrical stimuli in zoospore behaviors are also discussed. Finally, with the molecular mechanism of sporulation in Phytophthora and Peronophythora is gradually being revealed, we propose some thoughts for the further research and provide the alternative strategy for plant protection against phytopathogenic oomycetes.
Complete genome sequences of two Escherichia coli clinical isolates from Egypt carrying mcr-1 on IncP and IncX4 plasmids
Authors: Soliman Ahmed M. , Ramadan Hazem , Yu Liansheng , Hisatsune Junzo , Sugai Motoyuki , Elnahriry Shimaa S. , Nariya Hirofumi , El-Domany Ramadan A. , Shimamoto Toshi , Jackson Charlene R. , Shimamoto Tadashi
Published: September 2022

Colistin is a last-resort antibiotic used in the treatment of multidrug resistant Gram-negative bacteria. However, the activity and efficacy of colistin has been compromised by the worldwide spread of the mobile colistin resistance genes (mcr-1 to mcr-10). In this study, two clinical Escherichia coli strains, named EcCAI51, and EcCAI73, harbored mcr-1, showed multidrug-resistant phenotypes (with colistin MIC = 4 μg/ml), and belonged to phylogroup D: multilocus sequence type 1011 (ST1011) and phylogroup A: ST744, respectively. Findings revealed the existence of mcr-1 gene on two conjugable plasmids, pAMS-51-MCR1 (∼122 kb IncP) and pAMS-73-MCR1 (∼33 kb IncX4), in EcCAI51, and EcCAI73, respectively. The mcr-1-pap2 element was detected in the two plasmids. Additionally, the composite transposon (ISApl1-IS5D-pap2-mcr-1-ISApl1) was identified only in pAMS-51-MCR1 suggesting the potential for horizontal gene transfer. The two strains carried from 16 to 18 different multiple acquired antimicrobial resistance genes (ARGs). Additionally, two different multireplicon virulence plasmids (∼117 kb pAMS-51-Vr and ∼226 kb pAMS-73-Vr) carrying the sit operon, the Salmochelin siderophore iroBCDE operon and other several virulence genes were identified from the two strains. Hierarchical clustering of core genome MLST (HierCC) revealed clustering of EcCAI73, and EcCAI51 with global E. coli lineages at HC levels of 50 (HC50) to 100 (HC100) core genome allelic differences. To the best of our knowledge, this study presented the first complete genomic sequences of mcr-1-carrying IncP and IncX4 plasmids from human clinical E. coli isolates in Egypt. In addition, the study illustrated the mcr-1 broad dissemination in diverse plasmids and dissimilar E. coli clones.
Serum creatinine to cystatin C ratio and cognitive function among middle-aged and older adults in China
Authors: Zhu Yueli , Tan Zhongju , Li Shumin , Zhu Feng , Qin Chengfan , Zhang Qin , Yang Yunmei
Published: September 2022

Background

The sarcopenia index (SI, serum creatinine/serum cystatin C × 100) is recently suggested to be a reliable marker for sarcopenia. It has been reported that sarcopenia is associated with poorer cognition. The purpose of this study was to evaluate the correlation between SI and cognitive function among middle-aged and older adults from the China Health and Retirement Longitudinal Study (CHARLS).

Materials and methods

A total of 6,442 participants ≥45 years of age were enrolled in this study from CHARLS between 2011 and 2012. Cognitive function was assessed by interview-based measurements, including orientation and attention, episodic memory, visuo-construction, and the total cognitive function. SI was calculated by serum creatinine (mg/dL)/cystatin C (mg/L) × 100. One-way analysis of variance (ANOVA) was used to compare the differences among groups divided according to SI quartiles by gender. Both linear and logistic regression models were applied to investigate the relationship between SI and cognitive function.

Results

After adjustment for potential confounders, we found SI was significantly and positively correlated with total cognitive function scores both in males and females [β = 0.014, 95% confidence interval (CI) 0.007 to 0.021, P < 0.001; β = 0.011, 95 CI% 0.003 to 0.018, P = 0.004; respectively]. Similarly, when the total cognitive function score was treated as a categorical variable according to quartiles in males and females, higher SI was related to better total cognitive function scores in both males and females [odds ratio (OR) = 1.147, 95% CI 1.028 to 1.279, P = 0.014; OR = 1.219, 95% CI 1.106 to 1.344, P < 0.001; respectively] following adjustment for confounders.

Conclusions

Lower sarcopenia index was correlated with a higher prevalence of cognitive impairment among middle-aged and older adults in China.
The cortical spectrum: A robust structural continuum in primate cerebral cortex revealed by histological staining and magnetic resonance imaging
Authors: John Yohan J. , Zikopoulos Basilis , García-Cabezas Miguel Ángel , Barbas Helen
Published: September 2022

High-level characterizations of the primate cerebral cortex sit between two extremes: on one end the cortical mantle is seen as a mosaic of structurally and functionally unique areas, and on the other it is seen as a uniform six-layered structure in which functional differences are defined solely by extrinsic connections. Neither of these extremes captures the crucial neuroanatomical finding: that the cortex exhibits systematic gradations in architectonic structure. These gradations have been shown to predict cortico-cortical connectivity, which in turn suggests powerful ways to ground connectomics in anatomical structure, and by extension cortical function. A challenge to widespread use of this concept is the labor-intensive and invasive nature of histological staining, which is the primary means of recognizing anatomical gradations. Here we show that a novel computational analysis technique can provide a coarse-grained picture of cortical variation. For each of 78 cortical areas spanning the entire cortical mantle of the rhesus macaque, we created a high dimensional set of anatomical features derived from captured images of cortical tissue stained for myelin and SMI-32. The method involved semi-automated de-noising of images, and enabled comparison of brain areas without hand-labeling of features such as layer boundaries. We applied multidimensional scaling (MDS) to the dataset to visualize similarity among cortical areas. This analysis shows a systematic variation between weakly laminated (limbic) cortices and sharply laminated (eulaminate) cortices. We call this smooth continuum the “cortical spectrum”. We also show that this spectrum is visible within subsystems of the cortex: the occipital, parietal, temporal, motor, prefrontal, and insular cortices. We compared the MDS-derived spectrum with a spectrum produced using T1- and T2-weighted magnetic resonance imaging (MRI) data derived from macaque, and found close agreement of the two coarse-graining methods. This suggests that T1w/T2w data, routinely obtained in human MRI studies, can serve as an effective proxy for data derived from high-resolution histological methods. More generally, this approach shows that the cortical spectrum is robust to the specific method used to compare cortical areas, and is therefore a powerful tool to understand the principles of organization of the primate cortex.
Microglial pyroptosis: Therapeutic target in secondary brain injury following intracerebral hemorrhage
Authors: Gu Lingui , Sun Mingjiang , Li Ruihao , Tao Yihao , Luo Xu , Zhang Xingyu , Yuan Ye , Xie Zongyi
Published: September 2022

Intracerebral hemorrhage (ICH) is a major cerebrovascular illness that causes substantial neurological sequelae and dysfunction caused by secondary brain injury (SBI), and there are no effective therapies to mitigate the disability. Microglia, the brain-resident macrophage, participates in the primary inflammatory response, and activation of microglia to an M1-like phenotype largely takes place in the acute phase following ICH. A growing body of research suggests that the pathophysiology of SBI after ICH is mediated by an inflammatory response mediated by microglial-pyroptotic inflammasomes, while inhibiting the activation of microglial pyroptosis could suppress the inflammatory cascade reaction, thus attenuating the brain injury after ICH. Pyroptosis is characterized by rapid plasma membrane disruption, followed by the release of cellular contents and pro-inflammatory mediators. In this review, we outline the molecular mechanism of microglial pyroptosis and summarize the up-to-date evidence of its involvement in the pathological process of ICH, and highlight microglial pyroptosis-targeted strategies that have the potential to cure intracerebral hemorrhage. This review contributes to a better understanding of the function of microglial pyroptosis in ICH and assesses it as a possible therapeutic target.
Increased movement-related signals in both basal ganglia and cerebellar output pathways in two children with dystonia
Authors: Hernandez-Martin Estefania , Arguelles Enrique , Liker Mark , Robison Aaron , Sanger Terence D.
Published: September 2022

The contribution of different brain regions to movement abnormalities in children with dystonia is unknown. Three awake subjects undergoing depth electrode implantation for assessments of potential deep brain recording targets performed a rhythmic figure-8 drawing task. Two subjects had dystonia, one was undergoing testing for treatment of Tourette Syndrome and had neither dystonia nor abnormal movements during testing. Movement-related signals were evaluated by determining the magnitude of task-related frequency components. Brain signals were recorded in globus pallidus internus (GPi), the ventral oralis anterior/posterior (VoaVop) and the ventral intermediate (Vim) nuclei of the thalamus. In comparison to the subject without dystonia, both children with dystonia showed increased task-related activity in GPi and Vim. This finding is consistent with a role of both basal ganglia and cerebellar outputs in the pathogenesis of dystonia. Our results further suggest that frequency analysis of brain recordings during cyclic movements may be a useful tool for analysis of the presence of movement-related signals in various brain regions.
A predictive hemodynamic model based on risk factors for ruptured mirror aneurysms
Authors: Hu Sheng-qi , Chen Ru-dong , Xu Wei-dong , Li Hua , Yu Jia-sheng
Published: September 2022

Objectives

To identify hemodynamic risk factors for intracranial aneurysm rupture and establish a predictive model to aid evaluation.

Methods

We analyzed the hemodynamic parameters of 91 pairs of ruptured mirror aneurysms. A conditional univariate analysis was used for the continuous variables. A conditional multivariate logistic regression analysis was performed to identify the independent risk factors. Differences where p < 0.05 were statistically significant. A predictive model was established based on independent risk factors. Odds ratios (ORs) were used to score points. The validation cohort consisted of 189 aneurysms. Receiver operating characteristic curves were generated to determine the cutoff values and area under the curves (AUCs) of the predictive model and independent risk factors.

Results

The conditional multivariate logistic analysis showed that the low shear area (LSA) (OR = 70.322, p = 0.044, CI = 1.112–4,445.256), mean combined hemodynamic parameter (CHP) (>0.087) (OR = 3.171, p = 0.034, CI = 1.089–9.236), and wall shear stress gradient (WSSG) ratio (>893.180) (OR = 5.740, p = 0.003, CI = 1.950–16.898) were independent risk factors. A prediction model was established: 23*LSA + 1*CHP mean (>0.087: yes = 1, no = 0) + 2 * WSSG ratio (>893.180: yes = 1, no = 0). The AUC values of the predictive model, LSA, mean CHP (>0.087), and WSSG ratio (>893.180) were 0.748, 0.700, 0.654, and 0.703, respectively. The predictive model and LSA cutoff values were 1.283 and 0.016, respectively. In the validation cohort, the predictive model, LSA, CHP (>0.087), and WSSG ratio (>893.180) were 0.736, 0.702, 0.689, and 0.706, respectively.

Conclusions

LSA, CHP (>0.087), and WSSG ratio (>893.180) were independent risk factors for aneurysm rupture. Our predictive model could aid practical evaluation.
A novel deep learning approach for diagnosing Alzheimer's disease based on eye-tracking data
Authors: Sun Jinglin , Liu Yu , Wu Hao , Jing Peiguang , Ji Yong
Published: September 2022

Eye-tracking technology has become a powerful tool for biomedical-related applications due to its simplicity of operation and low requirements on patient language skills. This study aims to use the machine-learning models and deep-learning networks to identify key features of eye movements in Alzheimer's Disease (AD) under specific visual tasks, thereby facilitating computer-aided diagnosis of AD. Firstly, a three-dimensional (3D) visuospatial memory task is designed to provide participants with visual stimuli while their eye-movement data are recorded and used to build an eye-tracking dataset. Then, we propose a novel deep-learning-based model for identifying patients with Alzheimer's Disease (PwAD) and healthy controls (HCs) based on the collected eye-movement data. The proposed model utilizes a nested autoencoder network to extract the eye-movement features from the generated fixation heatmaps and a weight adaptive network layer for the feature fusion, which can preserve as much useful information as possible for the final binary classification. To fully verify the performance of the proposed model, we also design two types of models based on traditional machine-learning and typical deep-learning for comparison. Furthermore, we have also done ablation experiments to verify the effectiveness of each module of the proposed network. Finally, these models are evaluated by four-fold cross-validation on the built eye-tracking dataset. The proposed model shows 85% average accuracy in AD recognition, outperforming machine-learning methods and other typical deep-learning networks.
Evaluation of in vitro and in vivo Glycemic Index of common staples made from varieties of White Yam (Dioscorea rotundata)
Authors: Eyinla Toluwalope Emmanuel , Sanusi Rasaki Ajani , Maziya-Dixon Busie
Published: September 2022

Consumption of high Glycemic Index (GI) foods is a risk factor for increasing prevalence of diabetes mellitus (DM). The extent of variation in starch digestibility and GI of Yam varieties and products is not yet fully understood. This study was therefore designed to evaluate in vitro and in vivo Glycemic Index properties of commonly consumed products prepared from varieties of White Yam. Four products (boiled, fried, pounded yam, and Amala) were prepared from 5 common varieties of Yam and evaluated for Digestible Starch (DS) and Resistant Starch (RS). Based on results, two products-pounded yam and Amala-were processed from three of the most popular varieties. Analysis of Rapidly Digestible Starch (RDS), Slowly Digestible Starch (SDS), and estimated in vitro GI (eGI) were then carried out in this stage. Glycemic Index (GI) of these products consumed by apparently healthy young adults were also determined. Variety Amula had highest DS in Amala (19.1/100 g) and pounded Yam (20.4/100 g) while variety Alumaco had highest RS in all the products (2.9–3.3/100 g). When compared with RS in its raw tuber, RS of Alumaco generally increased after processing. Variety Alumaco had lowest RDS in Amala (0.6/100 g) and pounded yam (0.3/100 g) while eGI was lowest in Alumaco made into Amala (53) and pounded yam (48). Assessment of GI resulted in high GI for all products across each variety. Irrespective of variety, processing Yam into Amala released RDS fraction faster when compared with pounded yam. Generally, even though the products are considered as having high GI, Amala raised eGI and GI faster than Pounded yam. Variety-Alumaco particularly showed favorable properties applicable to dietary management of diabetes. Exploring more processing methods and genetic diversity is recommended.
Personalized nutrition: A review of genotype-based nutritional supplementation
Authors: Wang Feijie , Zheng Jianheng , Cheng Junrui , Zou Hong , Li Mingfeng , Deng Bin , Luo Rong , Wang Feng , Huang Dingqiang , Li Gang , Zhang Rao , Ding Xin , Li Yuan , Du Jun , Yang Yuexin , Kan Juntao
Published: September 2022

Nutritional disorders have become a major public health issue, requiring increased targeted approaches. Personalized nutrition adapted to individual needs has garnered dramatic attention as an effective way to improve nutritional balance and maintain health. With the rapidly evolving fields of genomics and nutrigenetics, accumulation of genetic variants has been indicated to alter the effects of nutritional supplementation, suggesting its indispensable role in the genotype-based personalized nutrition. Additionally, the metabolism of nutrients, such as lipids, especially omega-3 polyunsaturated fatty acids, glucose, vitamin A, folic acid, vitamin D, iron, and calcium could be effectively improved with related genetic variants. This review focuses on existing literatures linking critical genetic variants to the nutrient and the ways in which these variants influence the outcomes of certain nutritional supplementations. Although further studies are required in this direction, such evidence provides valuable insights for the guidance of appropriate interventions using genetic information, thus paving the way for the smooth transition of conventional generic approach to genotype-based personalized nutrition.
Real-world experience with anti–programmed cell death protein 1 immunotherapy in patients with esophageal cancer: A retrospective single-center study
Authors: Wang Xinpeng , Cai Lvjuan , Wu Mengjing , Li Guo , Zhu Yunyun , Lin Xinyue , Yan Xue , Mo Peng , Luo Huachun , Fu Zhichao
Published: September 2022

The “real-world” data of programmed cell death protein 1 (PD-1) inhibitors in esophageal cancer (EPC) are still an unmet medical need, including the clinical efficacy and safety. Seventy-seven EPC data were studied retrospectively; the progression-free survival (PFS), risk factors (clinical stages larger than stage II, metastatic sites larger than 2, treatment lines larger than the first line, previous surgical treatment, combined positive score [CPS] expression, etc.), and the safety were analyzed. The median PFS for all patients was 7.2 months, clinical stage > stage II; the number of treatment lines > first line was significantly correlated with prognosis (all P < 0.05). Subgroup analysis showed that the median PFS of patients with clinical stage ≤ II was better; the results were the same for the patients with ≤2 metastatic sites, first-line PD-1 inhibitors, and not previously received radical surgery (all P < 0.05). Meanwhile, the incidence of adverse events (AEs) of varying degrees was 25.97% (20/77) in 20 patients and 6.49% (5/77) of grade 3/4 AEs. The highest AE was myelosuppression (15.58%), followed by liver function injury (7.79%). In addition, ≥2 lines of treatment and >2 metastatic sites predicted poor outcomes for patients with EPC who had failed first-line therapy or progressed with the combined immunotherapy and chemotherapy treatment strategy (all P < 0.05).
Flow-controlled ventilation maintains gas exchange and lung aeration in a pediatric model of healthy and injured lungs: A randomized cross-over experimental study
Authors: Schranc Álmos , Balogh Ádám L. , Diaper John , Südy Roberta , Peták Ferenc , Habre Walid , Albu Gergely
Published: September 2022

Flow-controlled ventilation (FCV) is characterized by a constant flow to generate active inspiration and expiration. While the benefit of FCV on gas exchange has been demonstrated in preclinical and clinical studies with adults, the value of this modality for a pediatric population remains unknown. Thus, we aimed at observing the effects of FCV as compared to pressure-regulated volume control (PRVC) ventilation on lung mechanics, gas exchange and lung aeration before and after surfactant depletion in a pediatric model. Ten anesthetized piglets (10.4 ± 0.2 kg) were randomly assigned to start 1-h ventilation with FCV or PRVC before switching the ventilation modes for another hour. This sequence was repeated after inducing lung injury by bronchoalveolar lavage and injurious ventilation. The primary outcome was respiratory tissue elastance. Secondary outcomes included oxygenation index (PaO2/FiO2), PaCO2, intrapulmonary shunt (Qs/Qt), airway resistance, respiratory tissue damping, end-expiratory lung volume, lung clearance index and lung aeration by chest electrical impedance tomography. Measurements were performed at the end of each protocol stage. Ventilation modality had no effect on any respiratory mechanical parameter. Adequate gas exchange was provided by FCV, similar to PRVC, with sufficient CO2 elimination both in healthy and surfactant-depleted lungs (39.46 ± 7.2 mmHg and 46.2 ± 11.4 mmHg for FCV; 36.0 ± 4.1 and 39.5 ± 4.9 mmHg, for PRVC, respectively). Somewhat lower PaO2/FiO2 and higher Qs/Qt were observed in healthy and surfactant depleted lungs during FCV compared to PRVC (p < 0.05, for all). Compared to PRVC, lung aeration was significantly elevated, particularly in the ventral dependent zones during FCV (p < 0.05), but this difference was not evidenced in injured lungs. Somewhat lower oxygenation and higher shunt ratio was observed during FCV, nevertheless lung aeration improved and adequate gas exchange was ensured. Therefore, in the absence of major differences in respiratory mechanics and lung volumes, FCV may be considered as an alternative in ventilation therapy of pediatric patients with healthy and injured lungs.
Newborns' clinical conditions are correlated with the neonatal assessment manual scorE (NAME)
Authors: Manzotti Andrea , Cerritelli Francesco , Lombardi Erica , La Rocca Simona , Biasi Pamela , Chiera Marco , Galli Matteo , Lista Gianluca
Published: September 2022

Objective

To investigate the relationship between the Neonatal Assessment Manual scorE (NAME) and newborns' clinical condition on a large number of infants. The NAME model was developed as an instrument to assess the infant's general conditions, especially in NICUs, by evaluating how the infant's body responds to an external stressor such as static touch. Previous studies, employing experienced assessors, showed good validity indices as well as high inter-rater reliability.

Study design

Newborns were recruited at the “Vittore Buzzi” Pediatric Hospital NICU ward in Milan and their clinical conditions were collected through a standardized form—the complexity index. Two manual practitioners assessed all eligible newborns using the NAME scores. Data was analyzed using Kendall's τ correlation and odds ratio (OR) to assess the relationship between the NAME scores and the complexity index.

Results

Two hundred two newborns (46% female; 34.1 w ± 4.3; birth weight of 2,093.4 gr ± 879.8) entered the study. The Kendall's correlation between the clinical conditions (complexity index) and the NAME score was −0.206 [95% CI: (−0.292, −0.116), p-value < 0.001], corresponding to an OR of 0.838 [95% CI: (0.757, 0.924), p-value < 0.001]. Further exploratory analyses showed significant correlation between gestational age, birth weight and NAME scores.

Conclusion

The present paper adds evidence to the NAME model validity by demonstrating its applicability in the clinical neonatological context.
Development of a novel Bruton’s tyrosine kinase inhibitor that exerts anti-cancer activities potentiates response of chemotherapeutic agents in multiple myeloma stem cell-like cells
Authors: Elbezanti Weam Othman , Al-Odat Omar S. , Chitren Robert , Singh Jaikee Kumar , Srivastava Sandeep Kumar , Gowda Krishne , Amin Shantu , Robertson Gavin P. , Nemmara Venkatesh V. , Jonnalagadda Subash C. , Budak-Alpdogan Tulin , Pandey Manoj K.
Published: September 2022

Despite recent improvements in multiple myeloma (MM) treatment, MM remains an incurable disease and most patients experience a relapse. The major reason for myeloma recurrence is the persistent stem cell-like population. It has been demonstrated that overexpression of Bruton’s tyrosine kinase (BTK) in MM stem cell-like cells is correlated with drug resistance and poor prognosis. We have developed a novel small BTK inhibitor, KS151, which is unique compared to other BTK inhibitors. Unlike ibrutinib, and the other BTK inhibitors such as acalabrutinib, orelabrutinib, and zanubrutinib that covalently bind to the C481 residue in the BTK kinase domain, KS151 can inhibit BTK activities without binding to C481. This feature of KS151 is important because C481 becomes mutated in many patients and causes drug resistance. We demonstrated that KS151 inhibits in vitro BTK kinase activities and is more potent than ibrutinib. Furthermore, by performing a semi-quantitative, sandwich-based array for 71-tyrosine kinase phosphorylation, we found that KS151 specifically inhibits BTK. Our western blotting data showed that KS151 inhibits BTK signaling pathways and is effective against bortezomib-resistant cells as well as MM stem cell-like cells. Moreover, KS151 potentiates the apoptotic response of bortezomib, lenalidomide, and panobinostat in both MM and stem cell-like cells. Interestingly, KS151 inhibits stemness markers and is efficient in inhibiting Nanog and Gli1 stemness markers even when MM cells were co-cultured with bone marrow stromal cells (BMSCs). Overall, our results show that we have developed a novel BTK inhibitor effective against the stem cell-like population, and potentiates the response of chemotherapeutic agents.
UGT2B7 gene polymorphism and linkage disequilibrium in pediatric epileptic patients and their influence on sodium valproate monotherapy: A cohort study
Authors: Adiga Sachidananda , PB Nandit , Adiga Usha , Shenoy Vijaya
Published: September 2022

Background: Uridine 5′-diphospho glucuronosyl transferase (UGT) is the main enzyme responsible for the glucuronide conjugation, the principal metabolic pathway of sodium valproate. The objective of the study was to explore if there was an association between the UGT2B7 genetic polymorphism and clinical efficacy and safety in paediatric epileptic patients on sodium valproate monotherapy.

Methods and materials: The cohort study included 100 pediatric epileptic patients aged 2–18 years who had been on sodium valproate monotherapy for at least 1 month. PCR-RFLP was carried out to assess the genetic polymorphism patterns of UGT2B7 (C161T, A268G, G211T). Based on the extent of seizure control throughout the 1-year follow-up, clinical outcome was assessed in terms of responders and non-responders. Hepatic, renal, and other lab parameters were assayed to determine safety. The SNPstat web software was used to calculate linkage disequilibrium.

Results: Out of 100 patients, CC (38%), CT (43%), TT (19%) pattern was observed in UGT2B7 (C161T) gene, AA (15%), AG (39%), GG (46%) in (A268G) gene and GG (80%), GT (18%), TT (02%) in (G211T) gene. There was no statistical difference in clinical outcome with distinct UGT2B7 genetic polymorphism patterns, according to the findings. With low D′ and R2 values, linkage disequilibrium between alleles was statistically insignificant. However, the associations of C161T and G211T with treatment response were significant (p = 0.014) in determining treatment response.

Conclusion: Our findings show that the genetic variation of UGT2B7 had no bearing on the clinical outcome of epilepsy. Gene interactions, on the other hand, had an impact on treatment response.
Mechanism underlying linezolid-induced peripheral neuropathy in multidrug-resistant tuberculosis
Authors: Yuan Yuan , Li Jinmeng , Chen Yanhong , Cai Qingshan , Xu Yingying , Lin Luting , Lang Yazhen , Guo Suhang , Zhang Ruoying , Cai Xinjun
Published: September 2022

Multidrug-resistant tuberculosis (MDR-TB) remains a main global health concern as there is no comprehensive therapeutic intervention yet and numerous adverse effects follow the therapeutic process. In recent years, linezolid has been frequently used for treating MDR-TB. However, peripheral neuropathy associated with linezolid has reduced patient compliance. The current study explored the mechanism underlying linezolid-induced peripheral neuropathy in MDR-TB. Autophagy plays a neuroprotective role against peripheral nerve injury. We hypothesized that autophagy might also play a neuroprotective role against linezolid-induced peripheral neuropathy. In this study, we collected 12 questionnaires from MDR-TB patients in our hospital, and 10 of them developed linezolid-induced pain. The pain is mainly concentrated in the feet and accompanied by numbness. Subsequently, we used Sprague-Dawley (SD) rats and Schwann cells (SCs) to explore the mechanism. We found that linezolid causes a sparse arrangement of sciatic nerve tissue with associated loss of neurons, myelin sheaths, and down-regulation of LC3B expression. These results were also confirmed by in vitro experiments, showing that linezolid inhibited the proliferation of SCs. And the expression of P-AKT and P62 was elevated, and the expression of LC3B declined compared with the control group. Moreover, chloroquine (CQ), an autophagy inhibitor, also exhibited experimental results similar to linezolid. In summary, we conclude that linezolid-induced peripheral neuropathy is associated with the inhibition of autophagy flux.
Fucoxanthin prevents breast cancer metastasis by interrupting circulating tumor cells adhesion and transendothelial migration
Authors: Wang Weiyu , Fu Chengbin , Lin Mengting , Lu Yusheng , Lian Shu , Xie Xiaodong , Zhou Guiyu , Li Wulin , Zhang Yiping , Jia Lee , Zhong Chunlian , Huang Mingqing
Published: September 2022

Metastasis is the leading cause of cancer-related death and a critical challenge in improving cancer treatment today. Circulating tumor cells (CTCs) adhesion to and across the vascular endothelium are critical steps in the establishment of micrometastatic foci away from the primary tumor. Therefore, we believe that interrupting CTCs adhesion to endothelium and transendothelial migration may efficiently prevent cancer metastasis. Fucoxanthin (Fx) is an algal carotenoid widely distributed in brown algae, macroalgae, and diatoms. Previous studies have found that Fx has various pharmacological activities, including antidiabetic, antioxidant, anti-inflammatory, anti-obesity, antimalarial, anticancer, and so on. However, it remains unclear whether Fx has a preventive effect on cancer metastasis. Here, we found that Fx interrupts breast cancer cells MCF-7 adhesion to endothelium and transendothelial migration, thus inhibiting CTCs-based pulmonary metastasis in vivo. The hetero-adhesion assay showed that Fx significantly inhibited the expression of inflammatory factor-induced cell adhesion molecules (CAMs) and the resulting adhesion between MCF-7 cells and endothelial cells. The wound-healing and transwell assays showed that Fx significantly inhibited the motility, invasion, and transendothelial migration abilities of MCF-7 cells. However, the same concentration of Fx did not significantly alter the cell viability, cell cycle, apoptosis, and ROS of breast cancer cells, thus excluding the possibility that Fx inhibits MCF-7 cell adhesion and transendothelial migration through cytotoxicity. Mechanistically, Fx inhibits the expression of CAMs on endothelial cells by inhibiting the NF-кB signaling pathway by down-regulating the phosphorylation level of IKK-α/β, IкB-α, and NF-кB p65. Fx inhibits transendothelial migration of MCF-7 cells by inhibiting Epithelial-to-mesenchymal transition (EMT), PI3K/AKT, and FAK/Paxillin signaling pathways. Moreover, we demonstrated that Fx significantly inhibits the formation of lung micrometastatic foci in immunocompetent syngeneic mouse breast cancer metastasis models. We also showed that Fx enhances antitumor immune responses by substantially increasing the subsets of cytotoxic T lymphocytes in the peripheral immune system. This new finding provides a basis for the application of Fx in cancer metastatic chemoprevention and suggests that interruption of the CTCs adhesion to endothelium and transendothelial migration may serve as a new avenue for cancer metastatic chemoprevention.
Mailuoshutong pill for varicocele-associated male infertility—Phytochemical characterisation and multitarget mechanism
Authors: Lv Dongfang , Ji Yun , Zhang Qian , Shi Zhuozhuo , Chen Tengfei , Zhang Chao , Wang Xiangyun , Ren Taotao , Gao Zhaowang , Zhong Chongfu
Published: September 2022

Background: Varicocele (VC) is a relatively common and treatable cause of male infertility. Mailuoshutong pill (MLST), a traditional Chinese patent medicine, is widely used for treating varicose vein disease, but the underlying mechanism of MLST on varicocele-associated male infertility is unclear.

Objective: To reveal the phytochemical characterisation and multitarget mechanism of MLST on varicocele-associated male infertility.

Methods: The components in MLST were determined using UHPLC-MS/MS. Through network analysis, we constructed the “Drug-Components-Targets-Disease” network and predicted the potential biological functions and signaling pathways of MLST. Finally, the therapeutic effects and potential mechanisms of MLST were discovered by pharmacological experiments.

Results: By network analysis, the “Drug-Components-Targets-Disease” network was constructed, 62 components such as apigenin, limonin, kaempferol, and obacunoic acid may be the main active components of MLST for varicocele-associated male infertility, 28 targets such as VEGFA, PIK3CA, AKT1, and MTOR are considered as hub targets, signaling pathways such as HIF-1, Estrogen, PI3K/Akt, and mTOR may be key pathways for MLST against varicocele-associated male infertility. Through pharmacological experiments, we found that MLST ameliorated VC-induced testicular atrophy. Further histomorphology showed that MLST reduced VC-induced damage to testicular spermatogonia and seminiferous tubule, while MLST reduced ROS and MDA levels and increased antioxidant enzymes (GSH, GSH-Px, SOD, and CAT) levels. TUNEL staining and immunofluorescence showed that MLST reduced VC-induced apoptosis in testicular tissue, decreased BAX, and increased BCL2. Western blot results showed that MLST decreased the phosphorylation of PI3K, AKT, and mTOR proteins, and decreased the expression of HIF1α.

Conclusion: The phytochemical characterisation and multitarget mechanism of MLST on varicocele-associated male infertility were discovered using network analysis and pharmacological experiments. We verified that MLST can inhibit the activation of the PI3K/Akt/mTOR signaling pathway, reduce the expression of HIF1α, and further attenuate VC-induced oxidative stress and apoptosis in the testis. These findings provide evidence for the therapeutic role of MLST in varicocele-associated male infertility.
Evaluation of biological mechanisms of artemisinin on bovine mammary epithelial cells by integration of network pharmacology and TMT-based quantitative proteomics
Authors: Tong Jinjin , Sun Yang , Wang Ziyue , Cui Defeng , Jiang Linshu
Published: September 2022

The sesquiterpene lactone, artemisinin, is a primary component of the medicinal plant Artemisia annua L., which has anti-inflammatory, antibacterial and antioxidant activities. However, the potential effects of artemisinin on the mammary gland of dairy cows and the underlying molecular mechanisms remain unclear. Here, we utilized systematic network pharmacology and proteomics to elucidate the mechanism by which artemisinin affects milk production and the proliferation of bovine mammary epithelial cells (BMECs). Nineteen bioactive compounds and 56 key targets were identified through database mining. To delineate the mechanism of artemisia’s activity, a protein-protein interaction network and integrated visual display were generated from bioinformatics assays to explore the relationships and interactions among the bioactive molecules and their targets. The gene ontology (GO) terms and kyoto encyclopedia of genes and genomes annotation suggested that the apoptotic process, cell division, p53 pathway, prolactin and PI3K-Akt pathways played vital roles in mammary gland development. Using proteomics analysis, we identified 122 up-regulated and 96 down-regulated differentially significant expressed proteins (DSEPs). The differentially significant expressed proteins had multiple biological functions associated with cell division, apoptosis, differentiation, and migration. Gene ontology enrichment analysis suggested that differentially significant expressed proteins may promote cell proliferation and regulate apoptosis in bovine mammary epithelial cells. Kyoto encyclopedia of genes and genomes pathway analysis indicated that several biological pathways, such as those involved in antigen processing and presentation, cell adhesion molecules and ribosomes, played significant roles in the effects of artemisinin on bovine mammary epithelial cells. These findings contribute to a comprehensive understanding of the mechanism by which artemisinin affects bovine mammary epithelial cells to improve mammary gland turnover by inducing cell proliferation and mammary gland development.
Alismatis Rhizoma methanolic extract—Effects on metabolic syndrome and mechanisms of triterpenoids using a metabolomic and lipidomic approach
Authors: Jia Li , Zhang Min , Wang Pengli , Wang Liming , Lei Peng , Du Ruijiao , Han Lifeng , Zhang Peng , Wang Yuefei , Jiang Miaomiao
Published: September 2022

Alismatis rhizoma is a traditional Chinese medicine. Studies have demonstrated that Alismatis rhizoma also has therapeutic effects on metabolic syndrome. However, the pharmacodynamic material basis and mechanism are still unclear. First, UHPLC/Q-Orbitrap MS was used to detect the chemical components of the Alismatis rhizoma extract, and 31 triterpenoids and 2 sesquiterpenes were preliminarily identified. Then, to investigate the mechanism of the Alismatis rhizoma extract on metabolic syndrome, a mouse model of metabolic syndrome induced by high-fructose drinks was established. The results of serum biochemical analysis showed that the levels of TG, TC, LDL-C, and UA after the Alismatis rhizoma extract treatment were markedly decreased. 1H-NMR was used to conduct non-targeted metabolomics studies. A total of 20 differential metabolites were associated with high-fructose–induced metabolic syndrome, which were mainly correlated with 11 metabolic pathways. Moreover, UHPLC/Q-Orbitrap MS lipidomics analysis found that a total of 53 differential lipids were screened out. The results showed that Alismatis rhizoma extract mainly reduces the synthesis of glycerophospholipid and ceramide and improves the secretion of bile acid. This study shows that the Alismatis rhizoma extract can treat metabolic syndrome mainly by inhibiting energy metabolism, amino acid metabolism, and regulating bile acid to reduce phospholipid content.
Methodological considerations in studying digestive system physiology in octopus: limitations, lacunae and lessons learnt
Authors: Andrews Paul L. R. , Ponte Giovanna , Rosas Carlos
Published: September 2022

Current understanding of cephalopod digestive tract physiology is based on relatively “old” literature and a “mosaic of data” from multiple species. To provide a background to the discussion of methodologies for investigating physiology we first review the anatomy of the cephalopod digestive tract with a focus on Octopus vulgaris, highlighting structure-function relationships and species differences with potential functional consequences (e.g., absence of a crop in cuttlefish and squid; presence of a caecal sac in squid). We caution about extrapolation of data on the digestive system physiology from one cephalopod species to another because of the anatomical differences. The contribution of anatomical and histological techniques (e.g., digestive enzyme histochemistry and neurotransmitter immunohistochemistry) to understanding physiological processes is discussed. For each major digestive tract function we briefly review current knowledge, and then discuss techniques and their limitations for the following parameters: 1) Measuring motility in vitro (e.g., spatiotemporal mapping, tension and pressure), in vivo (labelled food, high resolution ultrasound) and aspects of pharmacology; 2) Measuring food ingestion and the time course of digestion with an emphasis on understanding enzyme function in each gut region with respect to time; 3) Assessing transepithelial transport of nutrients; 4) Measuring the energetic cost of food processing, impact of environmental temperature and metabolic rate (flow-through/intermittent respirometry); 4) Investigating neural (brain, gastric ganglion, enteric) and endocrine control processes with an emphasis on application of molecular techniques to identify receptors and their ligands. A number of major knowledge lacunae are identified where available techniques need to be applied to cephalopods, these include: 1) What is the physiological function of the caecal leaflets and intestinal typhlosoles in octopus? 2) What role does the transepithelial transport in the caecum and intestine play in ion, water and nutrient transport? 3) What information is signalled from the digestive tract to the brain regarding the food ingested and the progress of digestion? It is hoped that by combining discussion of the physiology of the cephalopod digestive system with an overview of techniques and identification of key knowledge gaps that this will encourage a more systematic approach to research in this area.
The microbiota changes of the brown dog tick, Rhipicephalus sanguineus under starvation stress
Authors: An Liping , Bhowmick Biswajit , Liang Dejuan , Suo Penghui , Liao Chenghong , Zhao Jianguo , Han Qian
Published: September 2022

Rhipicephalus sanguineus, the brown dog tick, is the most widespread tick in the world and a predominant vector of multiple pathogens affecting wild and domestic animals. There is an increasing interest in understanding the role of tick microbiome in pathogen acquisition and transmission as well as in environment–vector interfaces. Several studies suggested that the tick microbial communities are under the influence of several factors including the tick species, dietary bloodmeal, and physiological stress. Compared with insects, very little of the microbial community is known to contribute to the nutrition of the host. Therefore, it is of significance to elucidate the regulation of the microbial community of Rh. Sanguineus under starvation stress. Starvation stress was induced in wild-type adults (1 month, 2 months, 4 months, 6 months) and the microbial composition and diversity were analyzed before and after blood feeding. After the evaluation, it was found that the microbial community composition of Rh. sanguineus changed significantly with starvation stress. The dominant symbiotic bacteria Coxiella spp. of Rh. sanguineus gradually decreased with the prolongation of starvation stress. We also demonstrated that the starvation tolerance of Rh. sanguineus was as long as 6 months. Next, Coxiella-like endosymbionts were quantitatively analyzed by fluorescence quantitative PCR. We found a pronounced tissue tropism in the Malpighian tubule and female gonad, and less in the midgut and salivary gland organs. Finally, the blood-fed nymphs were injected with ofloxacin within 24 h. The nymphs were allowed to develop into adults. It was found that the adult blood-sucking rate, adult weight after blood meal, fecundity (egg hatching rate), and feeding period of the newly hatched larvae were all affected to varying degrees, indicating that the removal of most symbiotic bacteria had an irreversible effect on it.
Gut microbiome modulation mediated by probiotics: Positive impact on growth and health status of Labeo rohita
Authors: Ghori Ifra , Tubassam Misbah , Ahmad Tanveer , Zuberi Amina , Imran Muhammad
Published: September 2022

The current study was targeted to determine the effect of probiotics on the growth, physiology, and gut microbiology of Labeo rohita fingerlings. One hundred and twenty fishes were divided into four dietary groups, each in triplicate for a feeding trial of 90 days. These treatments included T0 (control, basal diet) used as the reference, and three probiotic-supplemented diets represented as Tbc (Bacillus cereus), Tgc (Geotrichum candidum), and Tmc (B. cereus and G. candidum). The probiotics were supplemented at a level of 1 × 109 CFU/g feed. Fishes nurtured on probiotic-added diet showed significantly high physiological improvement (p < 0.05) in terms of growth, feed utilization capacity, hematological profile, and digestive enzymes as compared to control. The fish were subjected to a challenge test after a 90-day feeding trial. The Tmc exhibited maximum fish growth when challenged by Staphylococcus aureus and showed fish survival when compared to control, in which fish mortality was examined. Fish gut microbial composition was modulated by probiotic treatments, especially in Tgc and Tmc as compared to control. The absence of opportunistic pathogens such as Staphylococcus saprophyticus and Sporobolomyces lactosus and detection of lower levels of Trichosporon and Cryptococcus in treated groups indicate the gut modulation driven by applied probiotics. The G. candidum QAUGC01 was retrieved in yeast metagenomics data, which might be due to the production of polyamines by them that facilitated adherence and consequent persistence. In conclusion, it can be suggested that the probiotic-supplemented diet could enhance fish growth and feed efficiency through community modulation and digestive enzymes, which could be a milestone in local aquaculture.