bluuebunny/crossref_metadata_2025 huggingface dataset:
Visualizing Cortical Development and Evolution: A Toolkit Update
Authors: Kumamoto Takuma , Ohtaka-Maruyama Chiaki
Published: April 2022

Visualizing the process of neural circuit formation during neurogenesis, using genetically modified animals or somatic transgenesis of exogenous plasmids, has become a key to decipher cortical development and evolution. In contrast to the establishment of transgenic animals, the designing and preparation of genes of interest into plasmids are simple and easy, dispensing with time-consuming germline modifications. These advantages have led to neuron labeling based on somatic transgenesis. In particular, mammalian expression plasmid, CRISPR-Cas9, and DNA transposon systems, have become widely used for neuronal visualization and functional analysis related to lineage labeling during cortical development. In this review, we discuss the advantages and limitations of these recently developed techniques.
Lipocalin-2 and Cerebral Stroke
Authors: Luo Chao , Zhou Shuai , Yin Shi , Jian Lipeng , Luo Pengren , Dong Jigeng , Liu Erheng
Published: April 2022

Stroke is a common and devastating disease with an escalating prevalence worldwide. The known secondary injuries after stroke include cell death, neuroinflammation, blood-brain barrier disruption, oxidative stress, iron dysregulation, and neurovascular unit dysfunction. Lipocalin-2 (LCN-2) is a neutrophil gelatinase-associated protein that influences diverse cellular processes during a stroke. The role of LCN-2 has been widely recognized in the peripheral system; however, recent findings have revealed that there are links between LCN-2 and secondary injury and diseases in the central nervous system. Novel roles of LCN-2 in neurons, microglia, astrocytes, and endothelial cells have also been demonstrated. Here, we review the evidence on the regulatory roles of LCN-2 in secondary injuries following a stroke from various perspectives and the pathological mechanisms involved in the modulation of stroke. Overall, our review suggests that LCN-2 is a promising target to promote a better understanding of the neuropathology of stroke.
Cephalopod Behavior: From Neural Plasticity to Consciousness
Authors: Ponte Giovanna , Chiandetti Cinzia , Edelman David B. , Imperadore Pamela , Pieroni Eleonora Maria , Fiorito Graziano
Published: April 2022

It is only in recent decades that subjective experience - or consciousness - has become a legitimate object of scientific inquiry. As such, it represents perhaps the greatest challenge facing neuroscience today. Subsumed within this challenge is the study of subjective experience in non-human animals: a particularly difficult endeavor that becomes even more so, as one crosses the great evolutionary divide between vertebrate and invertebrate phyla. Here, we explore the possibility of consciousness in one group of invertebrates: cephalopod molluscs. We believe such a review is timely, particularly considering cephalopods' impressive learning and memory abilities, rich behavioral repertoire, and the relative complexity of their nervous systems and sensory capabilities. Indeed, in some cephalopods, these abilities are so sophisticated that they are comparable to those of some higher vertebrates. Following the criteria and framework outlined for the identification of hallmarks of consciousness in non-mammalian species, here we propose that cephalopods - particularly the octopus - provide a unique test case among invertebrates for examining the properties and conditions that, at the very least, afford a basal faculty of consciousness. These include, among others: (i) discriminatory and anticipatory behaviors indicating a strong link between perception and memory recall; (ii) the presence of neural substrates representing functional analogs of thalamus and cortex; (iii) the neurophysiological dynamics resembling the functional signatures of conscious states in mammals. We highlight the current lack of evidence as well as potentially informative areas that warrant further investigation to support the view expressed here. Finally, we identify future research directions for the study of consciousness in these tantalizing animals.
External Validation of a Predictive Model for Acute Skin Radiation Toxicity in the REQUITE Breast Cohort
Authors: Rattay Tim , Seibold Petra , Aguado-Barrera Miguel E. , Altabas Manuel , Azria David , Barnett Gillian C. , Bultijnck Renée , Chang-Claude Jenny , Choudhury Ananya , Coles Charlotte E. , Dunning Alison M. , Elliott Rebecca M. , Farcy Jacquet Marie-Pierre , Gutiérrez-Enríquez Sara , Johnson Kerstie , Müller Anusha , Post Giselle , Rancati Tiziana , Reyes Victoria , Rosenstein Barry S. , De Ruysscher Dirk , de Santis Maria C. , Sperk Elena , Stobart Hilary , Symonds R. Paul , Taboada-Valladares Begoña , Vega Ana , Veldeman Liv , Webb Adam J. , West Catharine M. , Valdagni Riccardo , Talbot Christopher J. ,   REQUITE consortium
Published: April 2022

Background: Acute skin toxicity is a common and usually transient side-effect of breast radiotherapy although, if sufficiently severe, it can affect breast cosmesis, aftercare costs and the patient's quality-of-life. The aim of this study was to develop predictive models for acute skin toxicity using published risk factors and externally validate the models in patients recruited into the prospective multi-center REQUITE (validating pREdictive models and biomarkers of radiotherapy toxicity to reduce side-effects and improve QUalITy of lifE in cancer survivors) study.

Methods: Patient and treatment-related risk factors significantly associated with acute breast radiation toxicity on multivariate analysis were identified in the literature. These predictors were used to develop risk models for acute erythema and acute desquamation (skin loss) in three Radiogenomics Consortium cohorts of patients treated by breast-conserving surgery and whole breast external beam radiotherapy (n = 2,031). The models were externally validated in the REQUITE breast cancer cohort (n = 2,057).

Results: The final risk model for acute erythema included BMI, breast size, hypo-fractionation, boost, tamoxifen use and smoking status. This model was validated in REQUITE with moderate discrimination (AUC 0.65), calibration and agreement between predicted and observed toxicity (Brier score 0.17). The risk model for acute desquamation, excluding the predictor tamoxifen use, failed to validate in the REQUITE cohort.

Conclusions: While most published prediction research in the field has focused on model development, this study reports successful external validation of a predictive model using clinical risk factors for acute erythema following radiotherapy after breast-conserving surgery. This model retained discriminatory power but will benefit from further re-calibration. A similar model to predict acute desquamation failed to validate in the REQUITE cohort. Future improvements and more accurate predictions are expected through the addition of genetic markers and application of other modeling and machine learning techniques.
Anchored Multiplex PCR Custom Melanoma Next Generation Sequencing Panel for Analysis of Circulating Tumor DNA
Authors: Diefenbach Russell J. , Lee Jenny H. , Stewart Ashleigh , Menzies Alexander M. , Carlino Matteo S. , Saw Robyn P. M. , Stretch Jonathan R. , Long Georgina V. , Scolyer Richard A. , Rizos Helen
Published: April 2022

Detection of melanoma mutations using circulating tumor DNA (ctDNA) is a potential alternative to using genomic DNA from invasive tissue biopsies. To date, mutations in the GC-rich TERT promoter region, which is commonly mutated in melanoma, have been technically difficult to detect in ctDNA using next-generation sequencing (NGS) panels. In this study, we developed a custom melanoma NGS panel for detection of ctDNA, which encompasses the top 15 gene mutations in melanoma including the TERT promoter. We analyzed 21 stage III and IV melanoma patient samples who were treatment-naïve or on therapy. The overall detection rate of the custom panel, based on BRAF/NRAS/TERT promoter mutations, was 14/21 (67%) patient samples which included a TERT C250T mutation in one BRAF and NRAS mutation negative sample. A BRAF or NRAS mutation was detected in the ctDNA of 13/21 (62%) patients while TERT promoter mutations were detected in 10/21 (48%) patients. Co-occurrence of TERT promoter mutations with BRAF or NRAS mutations was found in 9/10 (90%) patients. The custom ctDNA panel showed a concordance of 16/21 (76%) with tissue based-detection and included 12 BRAF/NRAS mutation positive and 4 BRAF/NRAS mutation negative patients. The ctDNA mutation detection rate for stage IV was 12/16 (75%) and for stage III was 1/5 (20%). Based on BRAF, NRAS and TERT promoter mutations, the custom melanoma panel displayed a limit of detection of ~0.2% mutant allele frequency and showed significant correlation with droplet digital PCR. For one patient, a novel MAP2K1 H119Y mutation was detected in an NRAS/BRAF/TERT promoter mutation negative background. To increase the detection rate to >90% for stage IV melanoma patients, we plan to expand our custom panel to 50 genes. This study represents one of the first to successfully detect TERT promoter mutations in ctDNA from cutaneous melanoma patients using a targeted NGS panel.
Utility of Biology-Guided Radiotherapy to De Novo Metastases Diagnosed During Staging of High-Risk Biopsy-Proven Prostate Cancer
Authors: Gaudreault Mathieu , Chang David , Hardcastle Nicholas , Jackson Price , Kron Tomas , Hanna Gerard G. , Hofman Michael S. , Siva Shankar
Published: April 2022

Background

Biology-guided radiotherapy (BgRT) uses real-time functional imaging to guide radiation therapy treatment. Positron emission tomography (PET) tracers targeting prostate-specific membrane antigen (PSMA) are superior for prostate cancer detection than conventional imaging. This study aims at describing nodal and distant metastasis distribution from prostate cancer and at determining the proportion of metastatic lesions suitable for BgRT.

Methods

A single-institution patient subset from the ProPSMA trial (ID ACTRN12617000005358) was analysed. Gross tumour volumes (GTV) were delineated on the CT component of a PSMA PET/CT scan. To determine the suitability of BgRT tracking zones, the normalized SUV (nSUV) was calculated as the ratio of SUVmax inside the GTV to the SUVmean of adjacent three-dimensional shells of thickness 5 mm/10 mm/20 mm as a measure of signal to background contrast. Targets were suitable for BgRT if (1) nSUV was larger than an nSUV threshold and (2) non-tumour tissue inside adjacent shell was free of PET-avid uptake.

Results

Of this cohort of 84 patients, 24 had at least one pelvic node or metastatic site disease, 1 to 13 lesions per patient, with a total of 98 lesions (60 pelvic nodes/38 extra-pelvic nodal diseases and haematogenous metastases). Target volumes ranged from 0.08 to 9.6 cm3 while SUVmax ranged from 2.1 to 55.0. nSUV ranged from 1.9 to 15.7/2.4 to 25.7/2.5 to 34.5 for the 5 mm/10 mm/20 mm shell expansion. Furthermore, 74%/68%/34% of the lesions had nSUV ≥ 3 and were free of PSMA PET uptake inside the GTV outer shell margin expansion of 5 mm/10 mm/20 mm. Adjacent avid organs were another lesion, bladder, bowel, ureter, prostate, and liver.

Conclusions

The majority of PSMA PET/CT-defined radiotherapy targets would be suitable for BgRT by using a 10-mm tracking zone in prostate cancer. A subset of lesions had adjacent non-tumour uptake, mainly due to the proximity of ureter or bladder, and may require exclusion from emission tracking during BgRT.
Development and Validation of a Four Adenosine-to-Inosine RNA Editing Site-Relevant Prognostic Signature for Assessing Survival in Breast Cancer Patients
Authors: Wan Jian , Chen Shizhen , Zhang Anqin , Liu Yiting , Zhang Yangyang , Li Qinghua , Yu Ziqi , Wan Yuwei , Yang Lei , Wang Qi
Published: April 2022

Background

Adenosine-to-inosine RNA editing (ATIRE) is increasingly being used to characterize cancer. However, no studies have been conducted to identify an ATIRE signature for predicting cancer survival.

Methods

Breast cancer (BRCA) samples with ATIRE profiles from The Cancer Genome Atlas were divided into training (n = 452) and internal validation cohorts (n = 311), and 197 additional BRCA patients were recruited as an external validation cohort. The ATIRE signature for BRCA overall survival (OS) and disease-free survival (DFS) were identified using forest algorithm analysis and experimentally verified by direct sequencing. An ATIRE-based risk score (AIRS) was established with these selected ATIRE sites. Significantly prognostic factors were incorporated to generate a nomogram that was evaluated using Harrell’s C-index and calibration plot for all cohorts.

Results

Seven ATIRE sites were revealed to be associated with both BRCA OS and DFS, of which four sites were experimentally confirmed. Patients with high AIRS displayed a higher risk of death than those with low AIRS in the training (hazard ratio (HR) = 3.142, 95%CI = 1.932–5.111), internal validation (HR = 2.097, 95%CI = 1.123–3.914), and external validation cohorts (HR = 2.680, 95%CI = 1.000–7.194). A similar hazard effect of high AIRS on DFS was also observed. The nomogram yielded Harrell’s C-indexes of 0.816 (95%CI = 0.784–0.847), 0.742 (95%CI = 0.684–0.799), and 0.869 (95%CI = 0.835–0.902) for predicting OS and 0.767 (95%CI = 0.708–0.826), 0.684 (95%CI = 0.605–0.763), and 0.635 (95%CI = 0.566–0.705) for predicting DFS in the three cohorts.

Conclusion

AIRS nomogram could help to predict OS and DFS of patients with BRCA.
Oncolytic Activity of a Chimeric Influenza A Virus Carrying a Human CTLA4 Antibody in Hepatocellular Carcinoma
Authors: Yang Hao , Lei Guanglin , Sun Fang , Cheng Jinxia , Yan Jin , Zhang Shaogeng , Yang Penghui
Published: April 2022

Oncolytic virotherapy belongs to a kind of active immunotherapy, which could trigger a potent antitumor immune response, showing great potential in clinical application. OVs could induce immune responses through the dual mechanisms of selective tumor killing without destroying normal tissues and induction of systemic antitumor immunity. In this study, we successfully rescued a chimeric oncolytic influenza virus carrying a human CTLA4 antibody in the background of the A/PR/8/34 (PR8) virus. The chimeric virus, called rFlu-huCTLA4, contained the heavy and light chains of the human CTLA4 antibody in the PB1 and PA segments of the PR8 virus, respectively. The first-generation hemagglutination (HA) titers of the rFlu-huCTLA4 virus ranged from 27 to 28, which could be passaged stably in specific pathogen-free (SPF) chicken embryos from P1 to P5. The morphology and size distribution of the chimeric virus were consistent with those of the wt influenza virus. The rFlu-huCTLA4 virus could effectively replicate in various cells in time- and dose-dependent manners. ELISA assay revealed that the secreted huCTLA4 antibody levels in chicken embryos increased gradually over time. Furthermore, MTS and crystal violet analysis showed that the selective cytotoxicity of the virus was higher in hepatocellular carcinoma cells (HepG2 and Huh7) than in normal liver cells (MIHA). In vivo experiments displayed that intratumoral injection with rFlu-huCTLA4 reduced tumor growth and increased the survival of mice compared with the PR8 group. More importantly, in the rFlu-huCTLA4 group, we found that CD4+ and CD8 +T cells were significantly increased in tumor-bearing BALB/c mice. Taken together, these findings demonstrated that the chimeric oncolytic virus rFlu-huCTLA4 could selectively destroy hepatocellular carcinoma cells in vitro and in vivo and may provide a promising clinical strategy for targeted immunotherapy of HCC with the oncolytic flu virus.
Psychological Impact of the COVID-19 Pandemic on Pregnant Women
Authors: Buonsenso Danilo , Malorni Walter , Turriziani Colonna Arianna , Morini Sofia , Sbarbati Martina , Solipaca Alessandro , Di Mauro Antonio , Carducci Brigida , Lanzone Antonio , Moscato Umberto , Costa Simonetta , Vento Giovanni , Valentini Piero
Published: April 2022

Objective

The aim of this study is to assess the impact of the COVID-19 pandemic on mental health, type of delivery, and neonatal feeding of pregnant women with or without SARS-CoV-2 infection during gestation.

Study Design

The study was conducted online, and anonymous survey was distributed to mothers that delivered during the COVID-19 pandemic.

Results

The survey was completed by 286 women, and 64 women (22.4%) had COVID-19 during pregnancy. Women that had SARS-CoV-2 infection during pregnancy or at time of delivery had a significantly higher probability of being separated from the newborn (p < 0.0001) and a significantly lower probability of breastfeeding (p < 0.0001). The Edinburg Postnatal Depression Scale, to assess if mothers had symptoms of postnatal depression, showed that items suggestive of postnatal depression were relatively frequent in the whole cohort. However, women with SARS-CoV-2 infection during pregnancy reported higher probability of responses suggestive of postnatal depression in eight out of 10 items, with statistically significant differences in three items.

Conclusion

The COVID-19 pandemic affected the type of delivery and breastfeeding of pregnant women, particularly when they had SARS-CoV-2 infection. This, in turn, had an impact on the psychological status of the interviewed mothers, aspects that could benefit of special support.
D-dimer: The Risk Factor of Children's Severe Mycoplasma Pneumoniae Pneumonia
Authors: Qiu Juan , Ge Jin , Cao Ling
Published: April 2022

Objective

Mycoplasma Pneumoniae (MP) is an important cause of community-acquired pneumonia in children, which can cause serious consequences. There has been some research into predicting Severe Mycoplasma Pneumoniae Pneumonia (SMPP) primarily focused on pre-treatment time by macrolide, pre-hospital course, CRP and LDH et.al. while seldom reporting on concoagulation status. We designed this retrospective study to compare the difference between SMPP and Non-severe MPP (NSMPP) with an attempt to find the risk factors, with a special focus on concoagulation status.

Method

We performed a retrospective study of 786 MPP patients who were hospitalized from January 1, 2016 to December 31, 2018, age ranging from 28 days to 18 years old. All patients were divided into SMPP group and NSMPP group. A univariate analysis was conducted between both groups. The factors with statistical differences were included in logistic regression analysis to summarize the predictors of SMPP. Next, the predictive value of each risk factor was calculated from the receiver operating characteristic curve (ROC curve). Patients who had D-dimer records were divided into the elevated D-dimer group (D-dimer > 308ug/L) and the control group (D-dimer ≤ 308ug/L), and the clinical manifestations were compared.

Results

There was no significant difference in gender, age, pre-treatment time by macrolide, the white blood cell counts (WBC), Fibrinogen (FIB), Activated Partial Prothrombin Time (APTT), Prothrombin Time (PT) and Thrombin Time (TT) between SMPP and NSMPP. Compared with NSMPP, the pre-hospital course of SMPP was longer (P < 0.05), the neutrophil ratio (N%), platelet Count (PLT), C-reactive Protein (CRP), Lactate Dehydrogenase (LDH) and D-dimer were significantly higher (P < 0.01). The binary logistic regression analysis showed that the N%, PLT, CRP, LDH and D-dimer were the key predictors for SMPP, the N% > 67%, OR = 3.233, PLT > 445 × 109 /L, OR = 2.589, LDH > 354U/L, OR = 4.335 and D-dimer level > 403 ug/L, OR = 7.316. The D-dimer possessed the best predictive value. The incidence of complications such as pleural effusion, myocardial and liver damage of MPP was higher in the elevated D-dimer group than that in the control group (P < 0.05).

Conclusion

The N%, PLT, CRP, LDH and D-dimer were risk factors for SMPP. D-dimer was the best predictor among them. MPP patients with D-dimer > 308ug/L had more complications such as pleural effusion, myocardial and liver damage. More attention should be given in the treatment for this group.
Neural Activity Correlates With Behavior Effects of Anti-Seizure Drugs Efficacy Using the Zebrafish Pentylenetetrazol Seizure Model
Authors: Milder Patrick C. , Zybura Agnes S. , Cummins Theodore R. , Marrs James A.
Published: April 2022

Approximately 30% of patients with epilepsy do not achieve adequate seizure control through current anti-seizure drugs and treatment methods. Therefore, a critical need exists to efficiently screen anti-seizure drugs to enhance our ability to tailor treatment protocols and improve patient outcomes. The zebrafish pentylenetetrazol (PTZ) seizure model has become an increasingly popular screening paradigm for novel anti-seizure compounds. However, previous research using this model was variable due to differing experimental methods. Here, we present a method that was optimized to improve reliability and reproducibility in our laboratory using this PTZ model to develop a more robust screening of anti-seizure drugs comparing behavior and neural activity. Our behavior assay, spanning 90 min using 10 mM PTZ on 7 days post fertilization zebrafish, provides a broad window to observe anti-seizure drug efficacy. To compare our method with previously published data, we tested carbamazepine, lamotrigine, and topiramate, which have been tested in previous PTZ zebrafish assays. In addition, we assessed the candidate anti-seizure compound GS967, which has not been previously tested in the zebrafish seizure model. We examined the efficacy of anti-seizure drugs by acute administration concurrent with PTZ application and by pretreatment prior to exposure with PTZ. Pretreatment permitted us to examine potential neuroprotection and determine whether treatment time affects anti-seizure drugs’ responses. As independent validation of anti-seizure drugs’ effects, we evaluated whether the anti-seizure drug efficacy in the behavioral assay correlated with neural activity measurements, using electroencephalogram (EEG) and calcium signaling using GCaMP. There was no significant difference in the reduction of PTZ-induced seizure behavior activity between the pretreatment groups and acute treatment groups. Acute treatment with anti-seizure drugs in the EEG and GCaMP assays from 15 to 30 min post-anti-seizure drug exposure revealed consistent results between behavioral, EEG, and GCaMP assays for two of the three anti-seizure drugs. Lamotrigine only reduced neural activity (EEG and GCaMP assays). Carbamazepine, topiramate, and GS967 reduced activity in all three assays. The findings show that EEG and GCaMP assays largely correlate with the behavior findings, helping us connect physiological and behavior responses to anti-seizure drug and better assess anti-seizure drug efficacy.
New Advances on Pathophysiology of Diabetes Neuropathy and Pain Management: Potential Role of Melatonin and DPP-4 Inhibitors
Authors: Busa Prabhakar , Kuthati Yaswanth , Huang Niancih , Wong Chih-Shung
Published: April 2022

Pre-diabetes and diabetes are growing threats to the modern world. Diabetes mellitus (DM) is associated with comorbidities such as hypertension (83.40%), obesity (90.49%), and dyslipidemia (93.43%), creating a substantial burden on patients and society. Reductive and oxidative (Redox) stress level imbalance and inflammation play an important role in DM progression. Various therapeutics have been investigated to treat these neuronal complications. Melatonin and dipeptidyl peptidase IV inhibitors (DPP-4i) are known to possess powerful antioxidant and anti-inflammatory properties and have garnered significant attention in the recent years. In this present review article, we have reviewed the recently published reports on the therapeutic efficiency of melatonin and DPP-4i in the treatment of DM. We summarized the efficacy of melatonin and DPP-4i in DM and associated complications of diabetic neuropathy (DNP) and neuropathic pain. Furthermore, we discussed the mechanisms of action and their efficacy in the alleviation of oxidative stress in DM.
Development of a Rat Model of Intra-Amniotic Inflammation via Ultrasound-Guided Administration of a Triggering Agent in the Gestational Sac to Enable Analysis of Individual Amniotic Fluid Samples
Authors: Stranik Jaroslav , Kacerovsky Marian , Sterba Martin , Andrys Ctirad , Abad Cilia , Staud Frantisek , Micuda Stanislav , Soucek Ondrej , Jacobsson Bo , Musilova Ivana
Published: April 2022

Objectives: To develop a rat model of intra-amniotic inflammation, characterized by the concentration of interleukin-6 in the amniotic fluid, induced by an ultrasound-guided transabdominal administration of lipopolysaccharide into individual gestational sacs.

Methods: An ultrasound-guided transabdominal intra-amniotic administration of lipopolysaccharide or phosphate-buffered saline (PBS) as control was performed in rats on embryonic day 18. Only accessible gestational sacs with precise recording of their positions were injected. Twenty-four hours later, individual amniotic fluid samples were collected from the gestational sacs of laparotomized animals. The gestational sacs were divided into four subgroups: (i) with lipopolysaccharide: injected gestational sacs from rats undergoing lipopolysaccharide administration; (ii) without lipopolysaccharide: non-injected gestational sacs from rats undergoing lipopolysaccharide administration; (iii) with PBS: injected gestational sacs from rats undergoing PBS administration; and (iv) without PBS: non-injected gestational sacs from rats undergoing PBS administration. The concentration of interleukin-6 in individual amniotic fluid samples was assessed using ELISA.

Results: In the group of five animals receiving lipopolysaccharide, 24 (33%) and 48 (77%) gestational sacs were and were not injected, respectively. The amniotic fluid was obtained from 21 (88%) injected and 46 (95%) non-injected sacs. In the control group of five animals receiving phosphate-buffered saline, 28 (35%) and 52 (75%) gestational sacs were and were not injected, respectively. The amniotic fluid was obtained from 18 (64%) injected and 50 (96%) non-injected sacs. No labor occurred, and only one fetal death was observed in a gestational sac injected with lipopolysaccharide. Differences in concentrations of interleukin-6 in the amniotic fluid were found among the subgroups of the gestational sacs (with lipopolysaccharide: median 762 pg/ml; without lipopolysaccharide: median 35.6 pg/ml; with PBS: median 35.6 pg/ml; and without PBS: median 35.6 pg/ml; p < 0.0001). Concentrations of interleukin-6 in the amniotic fluid from the gestational sacs with lipopolysaccharide were significantly higher than those in the three remaining subgroups (p < 0.0001). No differences in concentrations of interleukin-6 in the amniotic fluid were identified between the three remaining subgroups.

Conclusion: The ultrasound-guided transabdominal intra-amniotic administration of lipopolysaccharide with a subsequent collection and analysis of amniotic fluid samples is feasible in rats. The intra-amniotic administration of lipopolysaccharide led to the development of intra-amniotic inflammation without leading to fetal mortality or induction of labor.
Cefiderocol for the Treatment of Multidrug-Resistant Gram-Negative Bacteria: A Systematic Review of Currently Available Evidence
Authors: Wang Chuanhai , Yang Deqing , Wang Yifan , Ni Wentao
Published: April 2022

Cefiderocol is a novel synthetic siderophore-conjugated antibiotic that hijacks the bacterial iron transport systems facilitating drug entry into cells, achieving high periplasmic concentrations. This systematic review analyzed the currently available literature on cefiderocol. It summarized in vitro susceptibility data, in vivo antimicrobial activity, pharmacokinetics/pharmacodynamics (PK/PD), clinical efficacy, safety and resistance mechanisms of cefiderocol. Cefiderocol has potent in vitro and in vivo activity against multidrug-resistant (MDR) Gram-negative bacteria, including carbapenem-resistant isolates. But New Delhi Metallo-β-lactamase (NDM)- positive isolates showed significantly higher MICs than other carbapenemase-producing Enterobacterales, with a susceptible rate of 83.4% for cefiderocol. Cefiderocol is well-tolerated, and the PK/PD target values can be achieved using a standard dose regimen or adjusted doses according to renal function. Clinical trials demonstrated that cefiderocol was non-inferiority to the comparator drugs in treating complicated urinary tract infection and nosocomial pneumonia. Case reports and series showed that cefiderocol was a promising therapeutic agent in carbapenem-resistant infections. However, resistant isolates and reduced susceptibility during treatment to cefiderocol have already been reported. In conclusion, cefiderocol is a promising powerful weapon for treating MDR recalcitrant infections.
Region-Dependent Increase of Cerebral Blood Flow During Electrically Induced Contraction of the Hindlimbs in Rats
Authors: Chaney Remi , Garnier Philippe , Quirié Aurore , Martin Alain , Prigent-Tessier Anne , Marie Christine
Published: April 2022

Elevation of cerebral blood flow (CBF) may contribute to the cerebral benefits of the regular practice of physical exercise. Surprisingly, while electrically induced contraction of a large muscular mass is a potential substitute for physical exercise to improve cognition, its effect on CBF remains to be investigated. Therefore, the present study investigated CBF in the cortical area representing the hindlimb, the hippocampus and the prefrontal cortex in the same anesthetized rats subjected to either acute (30 min) or chronic (30 min for 7 days) electrically induced bilateral hindlimb contraction. While CBF in the cortical area representing the hindlimb was assessed from both laser doppler flowmetry (LDFCBF) and changes in p-eNOSSer1177 levels (p-eNOSCBF), CBF was evaluated only from changes in p-eNOSSer1177 levels in the hippocampus and the prefrontal cortex. The contribution of increased cardiac output and increased neuronal activity to CBF changes were examined. Stimulation was associated with tachycardia and no change in arterial blood pressure. It increased LDFCBF with a time- and intensity-dependent manner as well as p-eNOSCBF in the area representing the hindlimb. By contrast, p-eNOSCBF was unchanged in the two other regions. The augmentation of LDFCBF was partially reduced by atenolol (a ß1 receptor antagonist) and not reproduced by the administration of dobutamine (a ß1 receptor agonist). Levels of c-fos as a marker of neuronal activation selectively increased in the area representing the hindlimb. In conclusion, electrically induced bilateral hindlimb contraction selectively increased CBF in the cortical area representing the stimulated muscles as a result of neuronal hyperactivity and increased cardiac output. The absence of CBF changes in cognition-related brain regions does not support flow-dependent neuroplasticity in the pro-cognitive effect of electrically induced contraction of a large muscular mass.
Influence of Step Frequency on the Dynamic Characteristics of Ventilation and Gas Exchange During Sinusoidal Walking in humans
Authors: Fujita Mako , Kamibayashi Kiyotaka , Aoki Tomoko , Horiuchi Masahiro , Fukuoka Yoshiyuki
Published: April 2022

We tested the hypothesis that restricting either step frequency (SF) or stride length (SL) causes a decrease in ventilatory response with limited breath frequency during sinusoidal walking. In this study, 13 healthy male and female volunteers (mean ± SD; age: 21.5 ± 1.8 years, height: 168 ± 7 cm, weight: 61.5 ± 8.3 kg) participated. The walking speed was sinusoidally changed between 50 and 100 m⋅min–1 with periods from 10 to 1 min. Using a customized sound system, we fixed the SF at 120 steps⋅min–1 with SL variation (0.83–0.41 m) (SFfix) or fixed the SL at 0.7 m with SF variation (143–71 steps⋅min–1) (SLfix) during the subjects’ sinusoidal walking. Both the subjects’ preferred locomotion pattern without a sound system (Free) and the unprompted spontaneous locomotor pattern for each subject (Free) served as the control condition. We measured breath-by-breath ventilation [tidal volume (VT) and breathing frequency (Bf)] and gas exchange [CO2 output (V.CO2), O2 uptake (V.O2)]. The amplitude (Amp) and the phase shift (PS) of the fundamental component of the ventilatory and gas exchange variables were calculated. The results revealed that the SFfix condition decreased the Amp of the Bf response compared with SLfix and Free conditions. Notably, the Amp of the Bf response under SFfix was reduced by less than one breath at the periods of 5 and 10 min. In contrast, the SLfix condition resulted in larger Amps of Bf and V.E responses as well as Free. We thus speculate that the steeper slope of the V.E-V.CO2 relationship observed under the SLfix might be attributable to the central feed-forward command or upward information from afferent neural activity by sinusoidal locomotive cadence. The PSs of the V.E, V.O2, and V.CO2 responses were unaffected by any locomotion patterns. Such a sinusoidal wave manipulation of locomotion variables may offer new insights into the dynamics of exercise hyperpnea.
Rotavirus-Induced Lipid Droplet Biogenesis Is Critical for Virus Replication
Authors: Criglar Jeanette M. , Estes Mary K. , Crawford Sue E.
Published: April 2022

A variety of pathogens, including viruses, bacteria and parasites, target cellular lipid droplets for their replication. Rotaviruses (RVs) infect the villous epithelium of the small intestine and are a major cause of acute gastroenteritis in infants and young children worldwide. RVs induce and require lipid droplets for the formation of viroplasms, sites of virus genome replication, and nascent particle assembly. Here we review the role of lipid droplets in RV replication. Inhibitors of fatty acid synthesis or chemicals that interfere with lipid droplet homeostasis decrease the number and size of viroplasms and the yield of infectious virus. We used a genetically engineered RV, delayed in viroplasm assembly, to show an early interaction of RV nonstructural protein NSP2 and the lipid droplet-associated protein phospho-PLIN1. The interaction between NSP2 and phospho-PLIN1 suggests that we have identified part of the mechanism of RV-induced lipid droplet formation. These studies demonstrate that RV is an excellent model to dissect the cellular process of lipid droplet formation and to determine how RV induces and usurps lipid droplet biogenesis to form viroplasm/lipid droplets for virus replication.
Weakly Supervised Deep Learning for Tooth-Marked Tongue Recognition
Authors: Zhou Jianguo , Li Shangxuan , Wang Xuesong , Yang Zizhu , Hou Xinyuan , Lai Wei , Zhao Shifeng , Deng Qingqiong , Zhou Wu
Published: April 2022

The recognition of tooth-marked tongues has important value for clinical diagnosis of traditional Chinese medicine. Tooth-marked tongue is often related to spleen deficiency, cold dampness, sputum, effusion, and blood stasis. The clinical manifestations of patients with tooth-marked tongue include loss of appetite, borborygmus, gastric distention, and loose stool. Traditional clinical tooth-marked tongue recognition is conducted subjectively based on the doctor’s visual observation, and its performance is affected by the doctor’s subjectivity, experience, and environmental lighting changes. In addition, the tooth marks typically have various shapes and colors on the tongue, which make it very challenging for doctors to identify tooth marks. The existing methods based on deep learning have made great progress for tooth-marked tongue recognition, but there are still shortcomings such as requiring a large amount of manual labeling of tooth marks, inability to detect and locate the tooth marks, and not conducive to clinical diagnosis and interpretation. In this study, we propose an end-to-end deep neural network for tooth-marked tongue recognition based on weakly supervised learning. Note that the deep neural network only requires image-level annotations of tooth-marked or non-tooth marked tongues. In this method, a deep neural network is trained to classify tooth-marked tongues with the image-level annotations. Then, a weakly supervised tooth-mark detection network (WSTDN) as an architecture variant of the pre-trained deep neural network is proposed for the tooth-marked region detection. Finally, the WSTDN is re-trained and fine-tuned using only the image-level annotations to simultaneously realize the classification of the tooth-marked tongue and the positioning of the tooth-marked region. Experimental results of clinical tongue images demonstrate the superiority of the proposed method compared with previously reported deep learning methods for tooth-marked tongue recognition. The proposed tooth-marked tongue recognition model may provide important syndrome diagnosis and efficacy evaluation methods, and contribute to the understanding of ethnopharmacological mechanisms.
Influences of Thermal Stress During Three Weeks Before Market Age on Histology and Expression of Genes Associated With Adipose Infiltration and Inflammation in Commercial Broilers, Native Chickens, and Crossbreeds
Authors: Malila Yuwares , Sanpinit Pornnicha , Thongda Wilawan , Jandamook Anuwat , Srimarut Yanee , Phasuk Yupin , Kunhareang Sajee
Published: April 2022

The objectives of this study were to examine the effects of cyclic thermal stress on histological characteristics of breast muscle and gene expression regarding adipose infiltration and inflammation in breast muscles collected from different breeds of chickens. The birds, from commercial broilers (CB, Ross 308, 3 weeks), native (NT, 100% Thai native Chee, 9 weeks), H75 (crossbred; 75% broiler and 25% NT, 5 weeks), and H50 (crossbred; 50% broiler and 50% NT, 7 weeks), were equally assigned into control or treatment groups. The control samples were reared under a constant temperature of 26 ± 1°C, while the treatment groups were exposed to 35 ± 1°C (6 h per day). After a 20-day thermal challenge, 12 male birds per treatment group were randomly collected for determination of live body weight, breast weight, numbers of growth-related myopathies, and breast meat chemical composition. Histological lesions were evaluated in the pectoralis major muscle immediately collected within 20 min postmortem based on hematoxylin and eosin staining. The results indicated that despite interaction between thermal stress and breed effects, thermal challenge significantly reduced feed intake, live body weight, and breast weight of the birds and increased moisture content in breast meat (p < 0.05). An interaction between the two main factors was found for protein content (p < 0.05) for which control CB showed less protein than the other groups. Heat stress decreased histological scores for adipose infiltration in CB (p < 0.05), but it did not significantly influence such scores in the other groups. CB received histological scores for adipose tissue at greater extent than those for the other groups. Differential absolute abundance of CD36, FABP4, LITAF, PDGFRA, PLIN1, PPARG, POSTN, SCD1, and TGFB1 in the muscle samples well-agreed with the trend of histological scores, suggesting potential involvement of dysregulated fibro-adipogenic progenitors together with imbalanced lipid storage and utilization in the breast muscle. The findings demonstrated that the cyclic thermal challenge restricted growth performance and breast mass of the birds, but such effects attenuated infiltration of adipose tissue and inflammatory cells in the CB breast muscle.
Non-Invasive Quantification of Fraction Flow Reserve Based on Steady-State Geometric Multiscale Models
Authors: Liu Jincheng , Wang Xue , Li Bao , Huang Suqin , Sun Hao , Zhang Liyuan , Sun Yutong , Liu Zhuo , Liu Jian , Wang Lihua , Zhao Xi , Wang Wenxin , Zhang Mingzi , Liu Youjun
Published: April 2022

Background: The underuse of invasive fraction flow reserve (FFR) in clinical practice has motivated research towards its non-invasive prediction. The early attempts relied on solving the incompressible three-dimensional Navier–Stokes equations in segmented coronary arteries. However, transient boundary condition has a high resource intensity in terms of computational time. Herein, a method for calculating FFR based on steady-state geometric multiscale (FFRSS) is proposed.

Methods: A total of 154 moderately stenotic vessels (40–80% diameter stenosis) from 136 patients with stable angina were included in this study to validate the clinical diagnostic performance of FFRSS. The method was based on the coronary artery model segmented from the patient’s coronary CTA image. The average pressure was used as the boundary condition for the inlet, and the microcirculation resistance calculated by the coronary flow was used as the boundary condition for the outlet to calculate the patient-specific coronary hyperemia. Then, the flow velocity and pressure distribution and the FFRss of each coronary artery branch were calculated to evaluate the degree of myocardial ischemia caused by coronary stenosis. Also, the FFRSS and FFRCT of all patients were calculated, and the clinically measured FFR was used as the “gold standard” to verify the diagnostic performance of FFRSS and to compare the correlation between FFRSS and FFRCT.

Results: According to the FFRSS calculation results of all patients, FFRSS and FFR have a good correlation (r = 0.68, p < 0.001). Similarly, the correlation of FFRSS and FFRCT demonstrated an r of 0.75 (95%CI: 0.67–0.72) (p < 0.001). On receiver-operating characteristic analysis, the optimal FFRSS cut point for FFR≤0.80 was 0.80 (AUC:0.85 [95% confidence interval: 0.79 to 0.90]; overall accuracy:88.3%). The overall sensitivity, specificity, PPV, and NPV for FFRSS ≤0.80 versus FFR ≤0.80 was 68.18% (95% CI: 52.4–81.4), 93.64% (95% CI: 87.3–97.4), 82.9%, and 91.1%, respectively.

Conclusion: FFRSS is a reliable diagnostic index for myocardial ischemia. This method was similar to the closed-loop geometric multiscale calculation of FFR accuracy but improved the calculation efficiency. It also improved the clinical applicability of the non-invasive computational FFR model, helped the clinicians diagnose myocardial ischemia, and guided percutaneous coronary intervention.
Contribution of Endocannabinoids to Intrinsic Motivation for Undirected Singing in Adult Zebra Finches
Authors: Kim Yunbok , Kojima Satoshi
Published: April 2022

Songbirds, such as zebra finches, spontaneously produce many song renditions for vocal practice even in the absence of apparent recipients throughout their lives. Such “undirected singing” is driven by intrinsic motivation, which arises within individuals for internal satisfaction without immediate external rewards. Intrinsic motivation for undirected singing in adult zebra finches was previously demonstrated to be critically regulated by dopamine through D2 receptors. Here, we further investigate the mechanisms of intrinsic motivation for undirected singing by focusing on endocannabinoids, which modulate dopamine signaling and contribute to motivation and reward in mammals. In songbirds, endocannabinoids have been shown to be involved in the production of undirected songs, but whether they are involved in the intrinsic motivation for undirected singing remains unknown. Using latencies of the first song production following temporary singing suppression as a measure of intrinsic motivation for undirected singing, we demonstrate that systemic administration of the direct cannabinoid agonist WIN55212-2 decreases intrinsic motivation for singing and that those effects are largely reversed by the cannabinoid antagonist SR141716A co-administered with WIN55212-2. Administration of SR141716A alone or that of two indirect cannabinoid agonists did not significantly affect intrinsic singing motivation. These results suggest that endocannabinoids are critically involved in regulating intrinsic motivation for undirected singing and provide new insights into the neural mechanisms of intrinsically motivated motor behaviors.
Loss and Natural Variations of Blast Fungal Avirulence Genes Breakdown Rice Resistance Genes in the Sichuan Basin of China
Authors: Hu Zi-Jin , Huang Yan-Yan , Lin Xiao-Yu , Feng Hui , Zhou Shi-Xin , Xie Ying , Liu Xin-Xian , Liu Chen , Zhao Ru-Meng , Zhao Wen-Sheng , Feng Chuan-Hong , Pu Mei , Ji Yun-Peng , Hu Xiao-Hong , Li Guo-Bang , Zhao Jing-Hao , Zhao Zhi-Xue , Wang He , Zhang Ji-Wei , Fan Jing , Li Yan , Peng Yun-Liang , He Min , Li De-Qiang , Huang Fu , Peng You-Liang , Wang Wen-Ming
Published: April 2022

Magnaporthe oryzae is the causative agent of rice blast, a devastating disease in rice worldwide. Based on the gene-for-gene paradigm, resistance (R) proteins can recognize their cognate avirulence (AVR) effectors to activate effector-triggered immunity. AVR genes have been demonstrated to evolve rapidly, leading to breakdown of the cognate resistance genes. Therefore, understanding the variation of AVR genes is essential to the deployment of resistant cultivars harboring the cognate R genes. In this study, we analyzed the nucleotide sequence polymorphisms of eight known AVR genes, namely, AVR-Pita1, AVR-Pii, AVR-Pia, AVR-Pik, AVR-Pizt, AVR-Pi9, AVR-Pib, and AVR-Pi54 in a total of 383 isolates from 13 prefectures in the Sichuan Basin. We detected the presence of AVR-Pik, AVR-Pi54, AVR-Pizt, AVR-Pi9, and AVR-Pib in the isolates of all the prefectures, but not AVR-Pita1, AVR-Pii, and AVR-Pia in at least seven prefectures, indicating loss of the three AVRs. We also detected insertions of Pot3, Mg-SINE, and indels in AVR-Pib, solo-LTR of Inago2 in AVR-Pizt, and gene duplications in AVR-Pik. Consistently, the isolates that did not harboring AVR-Pia were virulent to IRBLa-A, the monogenic line containing Pia, and the isolates with variants of AVR-Pib and AVR-Pizt were virulent to IRBLb-B and IRBLzt-t, the monogenic lines harboring Pib and Piz-t, respectively, indicating breakdown of resistance by the loss and variations of the avirulence genes. Therefore, the use of blast resistance genes should be alarmed by the loss and nature variations of avirulence genes in the blast fungal population in the Sichuan Basin.
Distribution Characteristics and Controlling Factors of Soil Total Nitrogen: Phosphorus Ratio Across the Northeast Tibetan Plateau Shrublands
Authors: Nie Xiuqing , Wang Dong , Ren Lining , Ma Kaili , Chen Yongzhe , Yang Lucun , Du Yangong , Zhou Guoying
Published: April 2022

Nitrogen (N) and phosphorus (P) stoichiometry have significant effects on nutrient cycles in terrestrial ecosystems. However, our understanding of the patterns and the driving factors of soil N:P ratios in the Tibetan Plateau shrublands remains limited. Our study aimed to quantify the distribution of soil N:P ratio and its controlling factors based on soil, plant, and climate factors from 59 sites in shrublands across the northeast Tibetan Plateau. The kriging interpolation method was used to quantify the soil N:P distribution. Spatially, the soil N:P ratio was higher in the south than in the north and lower in the west than in the east. The soil N:P ratio in the northeast Tibetan Plateau shrublands was mainly explained by edaphic factors, which also played an important role in regulating the effects of plant and climate factors on soil N:P ratios. Mean annual precipitation, instead of mean annual temperature, significantly controlled the soil N:P ratios, and its effect on the pattern of soil N:P ratios differed between alpine shrublands and desert shrublands. The N:P ratios of different organs in shrublands also played different roles in shaping the soil N:P ratios in alpine and desert shrublands. These results provide support for the hypothesis that edaphic factors were the dominant drivers of spatial variation in soil N:P ratios across the northeast Tibetan Plateau shrublands, and our study contributes to a deeper understanding of biogeochemical cycling at high altitudes.