bluuebunny/crossref_metadata_2025 huggingface dataset:
Determinantes da Estrutura de Capital das Empresas Listadas na Bolsa de Valores do Brasil: uma Análise Empírica do Período de 2007 ao 2016
Authors: Rabelo Henrique Marcelo , Braz Silva Sandro , Saporito Antonio
Published: July 2021

Esta pesquisa refere-se aos determinantes da estrutura de capital, utilizando a técnica de regressão múltipla por meio de dados em painel, de empresas de capital aberto nas bolsas de valores brasileiras, a fim de conhecer o comportamento dos determinantes da estrutura de capital em relação à Teoria do Trade-Off (TOT) e a Teoria da Ordem do Pecking (POT). Foram utilizadas dezesseis variáveis contábeis, das quais cinco são dependentes (relacionadas ao endividamento) e onze são variáveis independentes (explicativas dos determinantes da estrutura de capital). Observa-se que, com a utilização do Painel de Dados, os determinantes que parecem ter influência mais acentuada nos níveis de endividamento das empresas são: liquidez corrente, tangibilidade, retorno aos acionistas, retorno sobre ativos, crescimento das vendas, crescimento dos ativos, market-to-book e risco comercial medido pela volatilidade dos lucros.
Treatment of Osteoarthritis with Autologous, micro fragmented Adipose Tissue: A study protocol for a randomized controlled trial
Authors: Mikkelsen Rasmus Kramer , Blønd Lars , Hölmich Lisbeth Rosenkrantz , Mølgaard Cecilie , Troelsen Anders , Hölmich Per , Barfod Kristoffer Weisskirchner
Published: July 2021

Background: Osteoarthritis is a destructive joint disease that leads to degeneration of cartilage and other morphological changes in the joint. No medical treatment currently exists that can reverse these morphological changes. Intra-articular injection with autologous, micro fragmented adipose tissue has been suggested to relieve symptoms.Methods/Design: The study is a blinded randomized controlled trial with patients allocated in a 1:1 ratio to 2 parallel groups. Patients suffering from pain and functional impairment due to osteoarthritis Kellgren-Lawrence grade 2 – 3 in the tibiofemoral joint are eligible for inclusion. The intervention group is treated with an intra-articular injection with autologous, micro fragmented adipose tissue prepared using the Lipogems® system. The control group receives an intra-articular injection with isotonic saline. In total 120 patients are to be included. The primary outcome is The Knee injury and Osteoarthritis Outcome Score (KOOS4) evaluated at 6 months. Secondary outcomes are KOOS at 3, 12 and 24 months, the Tegner activity score, treatment failure and work status of the patient. The analysis will be conducted both as intention-to-treat and per-protocol analysis. Discussion: This trial is the first to investigate the efficacy of autologous, micro fragmented adipose tissue in a randomized controlled trial. The study uses the patient reported outcome measure Knee Injury and Osteoarthritis Outcome Score (KOOS4) after 6 months as the primary outcome, as it is believed to be a valid measure to assess the patient’s opinion about their knee and associated problems when suffering from osteoarthritis.
Physical Protection in Aggregates and Organo-Mineral Associations Contribute to Carbon Stabilization at the Transition Zone of Seasonally Saturated Wetlands
Authors: Kottkamp Anna , Jones C Nathan , Palmer Margaret A. , Tully Katherine L.
Published: July 2021

Wetlands store significant soil organic carbon (SOC) globally due to anoxic conditions that suppress SOC loss, yet this SOC is sensitive to climate and land use change. Seasonally saturated wetlands experience fluctuating hydrologic conditions that may also promote mechanisms known to control SOC stabilization in upland soils; these wetlands are therefore likely to be important for SOC storage at the landscape-scale. We investigated the role of physicochemical mechanisms of SOC stabilization in five seasonally saturated wetlands to test the hypothesis that these mechanisms are present, particularly in the transition between wetland and upland where soil saturation is most variable. At each wetland, we monitored water level and collected soil samples at five points along a transect from frequently saturated basin edge to rarely saturated upland. We quantified physical protection of SOC in aggregates and organo-mineral associations in mineral horizons to 0.5 m depth. As expected, SOC decreased from basin edge to upland. In the basin edge and transition zone, the majority of SOC was physically protected in macroaggregates. By contrast, overall organo-mineral associations were low, with the highest Fe concentrations (5 mg Fe g -1 soil) in the transition zone. While both stabilization mechanisms were present in the transition zone, physical protection is more likely to influence SOC stabilization during dry periods in seasonally saturated wetlands. As future climate scenarios predict changes in wetland wet and dry cycles, understanding the mechanisms by which SOC is stabilized in wetland soils is critical for predicting the vulnerability of SOC to future change.
GM-NAINO: Global Mutation based Novel Artificial Immune Network Optimization for Enhancing Data Security in Public Cloud Storage System
Authors: Kaliyamoorthy Priyadharshini , Ramalingam Aroul Canessane
Published: July 2021

The authors have requested that this preprint be removed from Research Square.
Impact of Lockdown During COVID-19 Outbreak on Ophthalmological Emergency in a Reference Center in France
Authors: MAALEJ Rim , HAGE Rabih , Salviat Flore , VIGNAL Catherine
Published: July 2021

PurposeIn March 2020, the sudden rise in number of SARS-CoV2 infections in France let the government to impose a strict lockdown during which all non-urgent medical consultations were postponed. From March 17 to May 10, 2020, private medical practices were closed and telemedicine was encouraged. The consequences on ophthalmic care were dramatic with over 90% of scheduled consultations canceled. The aim of this study was to report the consultation during the 2-month strict lockdown in Paris and to analyze its impact on the visual outcomes of patients consulting in the ophthalmology emergency department (OED) Methods :Data of patients who presented to the OED of the A. de Rothschild Foundation Hospital (RFH), a tertiary ophthalmology center in Paris, France, during the lockdown period and its immediate aftermath were analyzed. The results were compared to the same time periods in the years of 2018 and 2019. Four time periods were defined and numbered chronologically: March 17 to May 10, 2018 (Period 1) ; March 17 to May 10, 2019 (Period 2); March 17 to May 10, 2020 (Period 3, the lockdown period); May 11 to June 9, 2020 (Period 4, the post-lockdown period)Results :The number of consultation was reduced by more than 50% during the lockdown period (n=2909 patients) and by 30% during the post-lockdown period (n=2622) when compared to periods 1 (n=7125) and 2 (n=8058) Even though LP4 saw an increase in the number of consulting patients, there was no raise in the rate of severe diseases (12.8% during LP3 vs 11.1% during LP4) and the proportion of patients who were padmitted was statistically similar (4.3% VS 3.6%). Neuro-ophthalmic -diseases were the most common during LP3 and LP4. Neovascular glaucoma were twice as common during post LP4 (p=0.08). We highlighted a significant increase in patients with a graft reject consulting in our OED during the post LP4 (p<0.001). Those results were likely related to a delay in the follow-up consultations due to the lockdown measures.ConclusionThe reduction in the number of consultations in our OED during the lockdown period did not focus mainly on minor emergencies and also affected severe ophthalmic diseases with no significant delay in consultation. More longitudinal and longer study is needed to confirm this and to analyze retrospectively the COVID-19 outbreak and lockdown effects.
The Effects of Theory-Based Intervention on Self-Care Ability and Complications in Female Patients with Cancer with Peripherally Inserted Central Catheters (PICCs)
Authors: Huang Rong , Wu Na , Wang Ye , Jiang Hui , Zhang Li , Wang Hao
Published: July 2021

Purpose To examine the effects of Orem’s self-care theory-based continuous nursing intervention on self-care ability and PICC related complications and adverse events rates of female patients with cancer in the intermission of chemotherapy.Methods A quasi-experimental study was adopted. Totally 130 female patients with cancer were recruited from October 2017 to March 2019 at a tertiary maternal hospital in Shanghai. They were randomly divided into the experimental group (n=65) and the control group (n=65). The experimental group accepted nursing intervention based on Orem’s self-care theory, which had five sessions, while the control group accepted routine intervention. The exercise of self-care agency (ESCA) and the rates of PICC related complications and adverse events were compared by Chi square test and two‐sample t test between two groups through IBM SPSS 20.0 statistics software.Results The results showed that before the intervention, the baseline of two groups did not differ significantly. After the intervention, four dimensions and total scores of ESCA in the experimental group were significantly improved compared with those in the control group (p < 0.05), and the rates of PICC related complications and adverse events were significantly decreased in the experimental group compared with those in the control group (p < 0.05).Conclusions Orem’s self-care theory-based continuous nursing intervention was effective in improving self-care ability and decreasing the rates of PICC related complications and adverse events. Consequently, using the intervention program for patients with PICC during the intermission of chemotherapy is recommended.
The Impact of A Procedural Nursing Intervention to Prevent Deep Vein Thrombosis in Patients after Laparoscopic Distal Pancreatectomy and Splenectomy: A Retrospective Cohort Study
Authors: Xu Shan , Xiao Li , Wang Min , Zeng Tieying
Published: July 2021

Purpose The purpose of this study was to investigate the effect of procedural nursing intervention to prevent deep vein thrombosis (DVT) in patients after laparoscopic distal pancreatectomy and splenectomy (LDPS).Methods We conducted a retrospective analysis of data from 80 patients with tumours of the pancreatic corpus or tail who underwent surgery at a high volume single centre. T-test and c2 test were performed for data analysis. Study variables included incidence of DVT after LDPS, coagulation function indexes, and patient satisfaction. Results For patients after LDPS, individuals receiving the procedural nursing intervention had a lower incidence of DVT compared to individuals receiving routine nursing care. There was no significant difference in Fibrinogen (FBI), D-Dimer(D-D), Activated Partial Thromboplastin Time (APTT), Prothrombin Time (PT), Thrombin Time (TT) between the two groups (P>0.05). Platelet Count (PLT) among individuals receiving the procedural nursing intervention were significantly lower than those receiving routine nursing care. The procedural nursing interventions were associated with higher patient satisfaction compared to routine care. Conclusions A procedural nursing intervention contributes to a lower incidence of DVT, lower PLT, higher satisfaction.
REMBRANDT: A High-Throughput Barcoded Sequencing Approach for COVID-19 Screening.
Authors: Palmieri Dario , Javorina Amanda , Siddiquii Jalal , Gardner Anne , Fries Anthony , Chapleau Richard , Starr Clarise , Fishel Richard , Miles wayne
Published: July 2021

The Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2), also known as 2019 novel coronavirus (2019-nCoV), is a highly infectious RNA virus. A percentage of patients develop coronavirus disease 2019 (COVID-19) after infection, whose symptoms include fever, cough, shortness of breath and fatigue. Acute and life-threatening respiratory symptoms are experienced by 10-20% of symptomatic patients, particularly those with underlying medical conditions. One of the main challenges in the containment of COVID-19 is the identification and isolation of asymptomatic/pre-symptomatic individuals. A number of molecular assays are currently used to detect SARS-CoV-2. Many of them can accurately test hundreds or even thousands of patients every day. However, there are presently no testing platforms that enable more than 10,000 tests per day. Here, we describe the foundation for the REcombinase Mediated BaRcoding and AmplificatioN Diagnostic Tool (REMBRANDT), a high-throughput Next Generation Sequencing-based approach for the simultaneous screening of over 100,000 samples per day. The REMBRANDT protocol includes direct two-barcoded amplification of SARS-CoV-2 and control amplicons using an isothermal reaction, and the downstream library preparation for Illumina sequencing and bioinformatics analysis. This protocol represents a potentially powerful approach for community screening of COVID 19 that may be modified for application to any infectious or non-infectious genome.
Integrative Analysis and Identification of an Excellent lncRNA Signature to Predict Prognosis in Patients with COAD
Authors: Chen ZhiHua , Lin YiLin , Lin SuYong , Gao Ji , Chen Shao-Qin
Published: July 2021

Backgroud: Tumour recurrence and metastasis lead to poor prognosis incolon cancer(COAD). Therefore We aimed to identify a lncRNA signature through an integrative analysis of copy number variation, mutation and transcriptome data to predict prognosis and explore its internal mechanism.Methods: The lncRNA expression profile were collected fromThe Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO). TCGA data was randomly divided 3:1 intotraining andtesting cohort. In the training, weperformed integrated analyses of three candidate lncRNA sets that correlated with prognosis, copy number variations and mutations to establish a signature through Cox regression analysis. The robustness was determined in the testing and GEO.Results: An 11-lncRNA signature that was significantly associated with prognosiswas constructed in the training (P<0.0001, HR=2.014) , And this signature was validated in the testing(P=0.0019, HR=3.374) and GSE17536(P=0.0076, HR=1.864). The signature is significantly related to MSI status and clinical prognostic factors. The prognostic-relatedrisk scores were significantly excellent than the other five models have been reported. Furthermore, GSEA suggested that the signature was involved in COAD development and metastasis-related pathways.Conclusions: We identifiedansignature has strong robustness and can stably predict the prognosis of COAD in different platformsand may be implicated in COAD pathogenesis and metastasis and applied clinically as a prognostic marker.
In Vivo Quantitative Assessment of Therapeutic Response to Bortezomib Therapy in Disseminated Animal Models of Multiple Myeloma With [18F]FDG and [64Cu]LLP2A PET
Authors: Ghai Anchal , Fettig Nikki , Fontana Francesca , DiPersio John , Rettig Mike , Neal Julie O , Achilefu Samuel , Shoghi Kooresh , Shokeen Monica
Published: July 2021

BackgroundMultiple myeloma (MM) is a disease of cancerous plasma cells. Current treatments have improved the survival rate; however, most MM patients relapse. Imaging based timely determination of therapeutic response is critical for improving outcomes in MM patients. Very late antigen-4 (VLA4) is over expressed in MM cells. Here, we evaluated [18F]FDG and VLA4 targeted [64Cu]LLP2A for quantitative PET imaging in MM models of variable VLA4 expression, and following bortezomib therapy.MethodsIn vitro and ex vivo VLA4 expression was evaluated by flow cytometry. Human MM cells, MM.1S-CG and U266-CG (CG: luciferase and green fluorescent protein), were injected intravenously in NOD-SCID gamma mice. Tumor progression was monitored by bioluminescence imaging (BLI). Treatment group received bortezomib (1mg/kg, twice/week) intraperitoneally. All cohorts (treated, untreated and no-tumor) were longitudinally imaged with [64Cu]LLP2A (2-3 MBq; Molar Activity: 44.14±1.40 MBq/nmol) and [18F]FDG (7.4-8.0 MBq) PET respectively.ResultsFlow cytometry confirmed high expression of CD49d in U266 cells (>99%) and moderate expression in MM.1S cells (~52%). BLI showed decrease in total body flux in treated mice. In MM.1S-CG untreated versus treated mice, [64Cu]LLP2A localized with a significantly higher SUVmean in spine (0.58 versus 0.31) and femur (0.72 versus 0.39) at week 4 post tumor inoculation. In U266-CG treated versus untreated mice, there was a 4-time percent [64Cu]LLP2A increase in spine at week 3. Compared to [64Cu]LLP2A, [18F]FDG PET detected treatment related changes at later time points.Conclusion[64Cu]LLP2A is a promising tracer for in vivo assessment of therapeutic response in disseminated models of MM.
Skp2 Inhibitor AAA-237 Exerts Anti-Tumor Effect on NSCLC by Inducing Mitochondria-Dependent Apoptosis and Senescence
Authors: Liu Jinyi , Zheng Xiangjin , Li Wan , Ren Liwen , Li Sha , Yang Yihui , Ge Binbin , Du Guanhua , Shi Jianyou , Wang Jinhua
Published: July 2021

Background: Lung cancer is by far the leading cause of cancer death worldwide, and 85% of patients are non-small cell lung cancer (NSCLC). NSCLC is still very difficult to treat. Skp2 functions as an oncogene and participates in the processes of NSCLC. Here, we disclose a novel Skp2 inhibitor, which is firstly synthesized by our group based on our previous computer virtual screening and biomolecular interaction analysis (Biacore) technology screening. Methods: To unveil antitumor activity and underlying molecular mechanism of AAA-237, cell proliferation, migration and invasion assays, apoptosis, cell cycle, senescence, and xenograft mouse models were used.Results: AAA-237 inhibited the proliferation of NSCLC cells. AAA-237 regulated the process of epithelial–mesenchymal transition (EMT) by Skp2/Twist axis. Furthermore, AAA-237 induced arresting of cell cycle at G0/G1 phase by regulating the Skp2-Cip/Kip and PI3K/Akt-FOXO1 signaling pathways. More importantly, we found that treatment of high concentration AAA-237 could induce apoptosis of NSCLC cells via the mitochondrial pathway whereas treatment of low concentration AAA-237 for long-term could induce senescence of NSCLC cells by DNA damage. Senolytic drug quercetin could selectively eliminate senescent cells induced by AAA-237. In A549 xenograft nude mice model, AAA-237 dose-dependently inhibited the growth of tumor by inducing apoptosis and senescence.Conclusions: Taken together, we propose that AAA-237 is a promising therapeutic agent of NSCLC, especially when combined with senolytic drug.
Co-Delivery of Doxorubicin and Conferone by Novel pH-Responsive β-Cyclodextrin Grafted Micelles Triggers Apoptosis of Metastatic Human Breast Cancer Cells
Authors: Rahmani Akram , Rahimi Fariborz , Iranshahi Mehrdad , Kahroba Houman , Zarebkohan Amir , Talebi Mehdi , Salehi Roya , Mousavi Hassan Zavvar
Published: July 2021

Adjuvant-aided combination chemotherapy is one of the most effective ways of cancer treatment by overcoming the multidrug resistance (MDR) and reducing the side-effects of anticancer drugs. In this study Conferone (Conf) was used as adjuvant in combination with Doxorubicin (Dox) for induction of apoptosis to MDA-MB-231 cells. Herein, the novel biodegradable amphiphilic β-cyclodextrin grafted poly maleate-co-PLGA was synthesized by thiol-ene addition and ring-opening process. Micelles obtained from novel copolymer showed exceptional properties such as small size of around 34.5 nm, CMC of 0.1 μg/mL, and cell internalization of around 100 % at 30 min. These novel engineered micelles were used for combination delivery of doxorubicin-conferone with high encapsulation efficiency of near 100 % for both drugs. Our results show that combination delivery of Dox and Conf to MDA-MB-231 cells had synergistic effects (CI<1). According to cell cycle and Annexin-V apoptosis analysis, Dox-Conf loaded micelle significantly induce tumor cell apoptosis (more than 98 % of cells population showed apoptosis at IC50 = 0.259 μg/mL). RT-PCR and western-blot tests show that Dox-Conf loaded βCD-g-PMA-co-PLGA micelle induced apoptosis via intrinsic pathway. Therefore, the unique design of multi-functional pH-sensitive micelles open a new perspective for the development of nanomedicines for combination chemo-adjuvant therapy against malignant cancer.
Interlinking of Peri-Urban Lakes Based on Water Quality and Yield Using GIS for Efficient Water Management in Chennai, India
Authors: Partheeban Pachaivannan , Anuradha Baskaran , Arunbabu Elangovan , Somasundaram Krishnan , Hariharasudhan Chandarasekaran
Published: July 2021

This paper aims at providing a solution for augmenting the runoff water from major lakes of Chennai and Thiruvallur district of Tamilnadu. Hence an attempt was made to save surplus water from Puzhal, Ayanambakkam, Koratur, Ambattur, and Retteri lakes have been proposed. To interlink Puzhal lake with Koratur lake alignment is considered as it passes through the road network, excluding built-up areas. Water quality analysis was also done with a minimum of three samples that were collected from each lake. Samples were tested for pH, Total dissolved solids, Total suspended Solids, and Chemical oxygen demand. The linking of the lakes is proposed based on the water quality assessment of each lake. This study suggests an efficient/optimal route or canal for interlinking lakes in a GIS environment. The elevation differences, slope and dimensions of the canal, and the possible best location to link the lakes/tanks were obtained through the spatial data in the GIS. This helps in an effective and optimized management of the water supply for Chennai city.
Pulsed Electromagnetic Fields Protect Against Brain Injury After Intracerebral Hemorrhage: Involvement Of Anti-Inflammatory Processes And Hematoma Clearance Via CD36
Authors: Yang Yuefan , Wang Pan , Liu Anlai , Wu Xiuquan , Yan Zedong , Dai Shuhui , Wei Jialiang , Li Xin , Luo Peng , Luo Erping
Published: July 2021

Background: Intracerebral hemorrhage causes high mortality and morbidity, but its therapy methods are limited. In the present study, pulsed electromagnetic fields (PEMFs) were demonstrated to have beneficial effects on an intracerebral hemorrhage (ICH) model. This study explored the effects and underlying mechanism of PEMFs in a mouse model of ICH and cultured BV2 cells.Methods: PEMFs were applied 24 hours after collagenase-induced ICH and 4 hours per day for seven consecutive days. The levels of proinflammatory factors were assessed by ELISA kits and western blotting. Hematoma volume was measured by histological analysis. The effects of PEMFs on phagocytosis of the erythrocytes was observed in cultured BV2 cells.Results: Seven days after ICH, the hematoma volume was significantly reduced in PEMF-treated animals compared to nontreated mice. We found that PEMFs decreased the hematoma volume and the levels of proinflammatory factors after ICH. To evaluate the mechanisms underlying the effects of PEMFs on ICH, we analyzed the transcriptome profile three days after ICH, and we found that PEMFs reversed the changes in inflammation-related pathways and alleviated neurological deficits. Furthermore, PEMFs enhanced the erythrophagocytosis of microglia via CD36. Thus, the PEMF-mediated promotion of neurological functions may at least partly involve anti-inflammatory processes and hematoma clearance.Conclusions: These results suggest that PEMF treatment is a possible adjuvant therapy for ICH patients.
Use of An In Silico Knowledge Discovery Approach To Translate Mechanistic Studies of Silver Nanoparticles-Induced Toxicity From In Vitro To In Vivo
Authors: Mao Bin-Hsu , Luo Yi-Kai , W Bour-Jr , Cheng Fong-Yu , Lee Yu-Hsuan , Yan Shian-Jang , Wang Ying-Jan
Published: July 2021

Background: Silver nanoparticles (AgNPs) are considered a double-edged sword that demonstrates beneficial and harmful effects depending on their dimensions and surface coating types. However, mechanistic reports regarding size- and coating-dependent effects of AgNPs remain inadequate in vitro and in vivo. In keeping with the 3Rs principles, we adopted an in silico decision tree-based knowledge-discovery-in-databases process to offer a multilayered view of AgNPs toxicity and translate our research from cell-based phenomenological observations to further in vitro and in vivo mechanistic explorations.Results: Cell viability assessment results was used to create a tree model for predicting toxicity evoked by four AgNP types: SCS, LCS, SAS, and LAS. This model ranked toxicity-relevant parameters in the order: dose > cell type > AgNP type ≥ exposure time. As suggested by the model, we chose a less responsive cell line to conduct further investigations. LCS among others was more capable of comprising viability of this cell line and could trigger higher levels of apoptosis and autophagy at a subcytotoxic dose. Even though at a cytotoxic dose, it was yet unable to evoke necrosis. Longer exposure to a noncytotoxic dose of LCS induced G2/M cell cycle arrest and senescence rather than eliciting apoptosis and autophagy. After a single intraperitoneal injection, SCS was found to be more toxic to mice than SAS, both of which could be deposited in various target organs (e.g., spleen, liver, and kidneys). Morphological observation, together with serum biochemical and histological analyses, indicated that AgNPs could produce pancreatic toxicity, apart from leading to hepatic inflammation. Conclusions: Our integrated in vitro, in silico, and in vivo study demonstrated that AgNPs could exert toxicity in dose-, cell/organ type- and particle type-dependent manners. More importantly, a single injection of lethal-dose AgNPs (i.e., SCS and SAS) could incur severe damage to pancreas and raise blood glucose levels at the early phase of exposure.
Flow Cytometry Method for Absolute Counting and Single-Cell Phenotyping of Mycobacteria
Authors: Barr David A. , Omollo Charles , Mason Mandy , Koch Anastasia , Wilkinson Robert J. , Lalloo David G. , Meintjes Graeme , Mizrahi Valerie , Warner Digby F. , Davies Gerry
Published: July 2021

Detection and accurate quantitation of viable Mycobacterium tuberculosis is fundamental to understanding mycobacterial pathogenicity, tuberculosis (TB) disease progression and outcomes; TB transmission; drug action, efficacy and drug resistance. Despite this importance, methods for determining numbers of viable bacilli are limited in accuracy and precision owing to inherent characteristics of mycobacterial cell biology – including the tendency to clump, and “differential” culturability – and technical challenges consequent on handling an infectious pathogen under biosafe conditions. We developed an absolute counting method for mycobacteria in liquid cultures using a bench-top flow cytometer, and the low-cost fluorescent dyes Calcein-AM (CA) and SYBR-gold (SG). During exponential growth CA + cell counts are highly correlated with CFU counts and can be used as a real-time alternative to simplify the accurate standardisation of inocula for experiments. In contrast to CFU counting, this method can detect and enumerate cell aggregates in samples, which we show are a potential source of variance and bias when using established methods. We show that CFUs comprise a sub-population of intact, metabolically active mycobacterial cells in liquid cultures, with CFU-proportion varying by growth conditions. A pharmacodynamic application of the flow cytometry method, exploring kinetics of fluorescent probe defined subpopulations compared to CFU is demonstrated. Flow cytometry derived Mycobacterium bovis BCG time-kill curves differ for rifampicin and kanamycin versus isoniazid and ethambutol, as do the relative dynamics of discrete morphologically-distinct subpopulations of bacilli revealed by this high-throughput single-cell technique.
Quantitation and Characterization of Cell-Free Mitochondrial DNA in Plasma by Deep Sequencing
Authors: Daly Grant T. , Pastukh Viktor M. , Tan Yong B. , Francis C. Michael , Aggen C. Zack , Groark S. Chris , Edwards Carson , Hamo Mohammad , Simmons Jon D. , Kutcher Matthew E. , Hartsell Emily M. , Dinwiddie Darrell L. , Turpin Zachary M. , Bass Hank W. , Gillespie Mark N. , Langley Raymond J.
Published: July 2021

The presence in plasma of mitochondrial DNA (mtDNA) fragments, a proinflammatory damage-associated molecular pattern (DAMP), is positively associated with outcomes of multiple human disorders. Because mtDNA comprises only a small percentage of the total DNA in plasma, qPCR is typically employed as an analytic strategy. However, this method provides little insight into sequence origins or other characteristics of circulating mtDNA fragments. Here we found that target bait-capture applied to plasma mtDNA derived from severely injured patients enriched recovery by ≈ 1400-fold, thus affording depth sufficient for NextGen sequence analysis. Our method excluded nuclear mitochondrial insertions (NUMTs) using a stringent alignment and filtering strategy which calls and quantifies both NUMT in the reference genome and polymorphic NUMTs. After enrichment, we sequenced 2,000–60,000x mean coverage over the mtDNA genome. Normalization of mtDNA abundance to NUMT coverage reduced batch variability. Two massively-transfused trauma patients and two non-transfused patients displayed time-dependent increases in mtDNA DAMP coverage and decreases in the mean fragment length, which were more pronounced in massively transfused patients. Finally, our approach enabled detection of low-frequency heteroplasmic variants. Collectively, these findings suggest that our target bait-capture, deep sequencing and attendant analytic protocols could provide unprecedented characterization of cell-free plasma mtDNA and NUMTs.
Faecal Microbiota Transplantation from Patients with Depression Into Rats Modulates Mood-Related Behaviour
Authors: Sørensen Suzette , Knudsen Julie , Michaelsen Thomas , Bundgaard-Nielsen Caspar , Nielsen René Ernst , Hjerrild Simon , Leutscher Peter , Wegener Gregers
Published: July 2021

Differences in gut microbiota composition have been observed in patients with major depressive disorder (MDD) compared to healthy individuals. Here, we investigated if faecal microbiota transplantation (FMT) from MDD patients into rats, could induce a depressive-like phenotype. We performed FMT from patients with MDD (FMT-MDD) and healthy individuals (FMT-Healthy) into male Flinders Sensitive Line (FSL) and Flinders Resistant Line (FRL) rats and assessed depressive-like behaviour. No behavioural differences were observed in the FSL rats. In FRL rats, the FMT-Healthy group displayed significantly lower depressive-like behaviour than the FMT-MDD group. However, there was no difference in behaviour between FMT-MDD FRL rats and negative controls, indicating that FMT-Healthy FRL rats received beneficial bacteria. We additionally found different taxa between the FMT-MDD and the FMT-Healthy FRL rats, which could be traced to the donors. Four taxa, three belonging to Ruminococcaceae and the Lachnospira genus, were significantly elevated in relative abundance in FMT-MDD rats, while Coprococcus was depleted. In this study, the FMT-MDD group was different from the FMT-Healthy group based on behaviour and intestinal taxa.
Decision-Making Analysis Based on Hesitant Fuzzy N-Soft ELECTRE-I Approach
Authors: Adeel Arooj , Akram Muhammad , Cagman Naim
Published: July 2021

For the formal expression of uncertain data, hesitant fuzzy set theory has established itself as a distinguished model because it has a broad use in multi-attribute decision-making problems. With the incorporation of features from N -soft sets, a useful framework referred to as hesitant fuzzy N -soft sets has acquired an even greater appeal. This model integrates and associates the hesitant environment with information regarding the existence of grades or star ratings. In this research article, we introduce a multi-attribute decision-making technique known as hesitant fuzzy N -soft ELECTRE-I, which computes the decision-maker assessments in an adjustable and formative manner. The proposed method also improves the robustness and accuracy of the decisions relying on grades or star ratings. Thus it lays a bedrock for subsequent analyses and applications. We justify the relevance and convenience of the proposed technique by testing it in actually existing scenarios. Finally, we give a comparison of this novel methodology with the HFNS-TOPSIS method.
Gender Influence on Asymptomatic Malaria Diagnosis
Authors: Meirinho Sofia , Gomes Matilde , Campos Ana , Baptista Duarte , Baptista Alberta , Sequeira Gil , Ferreira Pedro Eduardo
Published: July 2021

The characterisation and identification of malaria reservoirs are major challenges for the efficacy of control and elimination programs. Malaria asymptomatic infections are highly prevalent in different malaria settings worldwide and constitute a great obstacle for disease control and elimination. The monitoring of asymptomatic malaria infection requires active case detection in many cases challenging the limit of diagnostic technology detection. We evaluated diagnostic methods and analyzed the gender dimension implication for asymptomatic malaria diagnosis on a high-endemic malaria region during the dry-season.
Continuum of Maternity Care Among Rural Women in Ethiopia: Does Place and Frequency of Antenatal Care Visit Matter?
Authors: Dadi Tegene Legese , Medhin Girmay , Kasaye Habtamu Kebebe , Kassie Getnet Mitike , Jebena Mulusew Gerbaba , Gobezie Wasihun Adualem , Alemayehu Yibeltal Kifle , Teklu Alula Meresa
Published: July 2021

Introduction: The ministry of health (MOH) of Ethiopia recommends 4 or more focused antenatal care (ANC) visits at health centre or at a higher level of health facility. In Ethiopia, few studies investigated time dimension of maternal health continuum of care but lack data regarding place dimension and its effect on continuum of care. The aim of this study is to estimate effect of place of ANC-1 visit and adherence to MOH’s recommendations of MOH for ANC visits on continuum of care rural in Ethiopia. Method: We used data collected from 1431 eligible women included in the National Health Extension Program (HEP) assessment survey that covered 6364 households from 62 woredas in nine regions. The main outcome variable is continuum of care (CoC), which is the uptake of all recommended ANC visits, institutional delivery and postnatal care services. Following descriptive analysis of the collected data, Propensity Score Matching was used to estimate the effect of place of ANC-1 visit on completion of CoC, and Zero inflated Poisson regression was used to model the effect of adherence to MOH recommendation of ANC visits on intensity of maternal health continuum of care. Result: Only 13.9% of eligible women completed the continuum of care and place of first antenatal care (ANC) visit was not significantly associated with the completion of continuum of care (β = 0.04, 95%CI= -0.02, 0.09). Adherence of ANC visit to the MOH recommendation (at least 4 ANC visits at higher HFs than HPs) increased the likelihood of higher intensity of continuum of care (aIRR = 1.29, 95%CI: 1.26, 1.33). Moreover, the intensity of continuum of care was positively associated with being in agrarian areas (aIRR = 1.17, 95%CI: 1.06, 1.29), exposed to HEP (IRR = 1.22, 95%CI: 1.16, 1.28), being informed about danger signs (aIRR = 1.14, 95%CI: 1.11, 1.18) and delivery of 2nd youngest child at HF (IRR = 1.16, 95%CI: 1.13, 1.20). Increasing age of women was negatively associated with use of services (IRR = 0.90, 95%CI: 0.87, 0.94). Conclusion: Completion of maternal health continuum of care is very low in Ethiopia, however most of the women use at least one of the services. Completion of continuum of care was not affected by place of first ANC visit. Adherence to MOH recommendation of ANC visit increased the intensity of continuum of care. Intensity of continuum of care was positively associated with residing in agrarian areas, HEP exposure, danger sign told, delivery of 2nd youngest child at health facility. To boost the uptake of all maternal health services, it is crucial to work on quality of health facilities, upgrading the infrastructures of HPs and promoting adherence to MOH recommendations of ANC visit.
Effect of a toe-walking protocol in plantar heel pain: A pilot study
Authors: Huffer Dean , Hing Wayne , Charles John , Newton Richard , Clair Mike , Canetti Elisa F. D.
Published: July 2021

Introduction/Purpose: Plantar heel pain (PHP) is one of most common disorders of the foot treated in primary care. It affects athletic and sedentary populations, with patient reports of activity-limiting pain and reduced quality of life. Recently, atrophy of the forefoot plantar intrinsic musculature was identified in patients with PHP.Therefore, the purpose of this study was to assess whether loading the plantar fascia strengthens the intrinsic foot musculature (IFM) and decreases PHP sufferers’ symptoms.Methods: A within-subjects experimental design assessed Foot Function Index (FFI) and IFM strength via hand-held dynamometry (HHD) in 12 patients with PHP prior to and at the end of a six-week toe-walking program.Results: After six weeks of treatment, the mean (SD) FFI score significantly decreased from 73.2 (32.4) to 43.3 (22.8) points (p = 0.010, ES = 1.1). HHD measures: Both great toe flexion and lesser toes flexion strength measures demonstrated significant force increase of 7.8N 95%CI [1.3, 14.4] (p = 0.024, ES = 1.0) and 6.6N 95%CI [1.8, 11.4] (p = 0.010, ES = 0.9), respectively.Conclusion: Results suggest toe-walking reduces PHP symptoms and increases IFM strength. There was no identified correlation between the IFM and FFI changes. Symptom reduction may potentially be due to a reduction in cortical inhibition rather than IFM strength changes. Findings provide foundation for future studies, employing imaging, to further validate the effectiveness of toe-walking in reducing symptoms of PHP patients.Trial registration: PROSPERO 2016 CRD42016036302 Registered 10 March 2016, https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42016036302
Relationship between self-rated health and depression risk among children: Promoting school–medical care cooperation for child suicide prevention
Authors: Okada Michiyo , Nakadoi Yoshihiro , Fujikawa Ai
Published: July 2021

Background

Japan is facing a markedly high incidence of adolescent suicide. This study examines the relationship between depression and self-rated health, which is a significant factor driving people toward suicide, highlighting the importance of children’s assessments of their health.
Methods

In this cross-sectional study, an original questionnaire combining text and illustrations was administered to 6,421 elementary, junior, and senior high school students. To assess children’s self-rated health, we classified the survey questions into two domains: physical and mental health. We further classified the questions based on symptom duration. Depression assessment was based on the Depression Self-Rating Scale for Children. To provide a basis for the effective use of the self-rated health assessment tool, we classified the respondents into three groups based on physical/mental conditions together with the duration of those conditions and made comparisons. The groups were Favorable Health, Temporarily Poor Health, and Persistently Poor Health, and the results were analyzed using an ANOVA.
Results

Self-rated health levels decreased, and depressive conditions worsened with age. Although most children led physically and mentally healthy school lives, the mean Depression Self-Rating Scale score was significantly higher for the Persistently Poor Health group than the other two, and the Temporarily Poor Health group scored significantly higher than the Favorable Health group for both physical and mental health (all p < 0.001).
Conclusions

As the Temporarily and Persistently Poor Health groups in the domains of physical and mental health are more likely to be depressed, timely cooperation between schools and medical institutions is imperative to prevent depression.
An Empirical Mode Decomposition Fuzzy Forecast Model for Air Quality
Authors: Chen Bo-Lun , Zhu Guo-Chang , Sheng Yi-Yun , Xie Qian , Ji Min , Yu Yong-Tao
Published: July 2021

Air quality is related to people's health. Severe air pollution can cause respiratory diseases, while good air quality is beneficial to physical and mental health. Therefore, the prediction of air quality is very important. As an important algorithm for signal analysis, empirical mode decomposition can analyze the change trend of air quality well, smooth the complex and changeable air quality data, and get the change trend of air quality under different time scales. According to the change trend under different time scales, the extreme learning machine is used for training, and the corresponding prediction value is obtained. The adaptive fuzzy inference system is used for fitting to obtain the final air quality prediction result. The experimental results show that the signal decomposition fuzzy prediction model has a good learning ability and has good accuracy in predicting the concentration of various pollutants in air quality.
Antibody-Mediated Blockade for Galectin-3 Binding Protein in Tumor Secretome Abrogates PDAC Metastasis
Authors: Choi Yeon-Sook , kim Myung Ji , Choi Eun A , Kim Sinae , Lee Eun Ji , Kim Hyun Ji , Jung Jae Yun , Kim Hyori , Kim Kyung Kon , Kim Jin Young , Hong Seung Mo , Kim Song Cheol , Chang Suhwan
Published: July 2021

Background

One of the major challenges in pancreatic ductal adenocarcinoma (PDAC) management is a local or distant metastasis and limited targeted therapeutics to prevent the process. We aimed to identify a druggable target by screening abnormally secreted protein from PDAC and explore its therapeutic intervention.
Methods

A LC-MS/MS-based proteomics was carried out for TIF (Tumor Interstitial Fluids) obtained from patient derived xenograft (PDX) models of PDAC. To develop a blocking antibody for selected target protein, antibody phage-display technology was used.
Results

The proteomic screening of PDAC secretome identified Galectin-3 binding protein (Gal-3BP hereafter) as a top candidate. The Gal-3BP is highly expressed and secreted in PDAC tumors and primary cells. Subsequent functional tests by stable knockdown revealed the Gal-3BP is required for PDAC cell proliferation, migration and invasion. In addition, the depletion of Gal-3BP significantly abrogated in vivo tumor formation and metastasis of pancreatic cancer cells. Mechanistically, we found Gal-3BP enhances the galectin-3 mediated EGFR signaling, leading to the activation of cMyc and its target genes related to EMT. To examine the clinical usability of these findings, we screened a Gal-3BP-immunized chicken antibody library using phage display technique. The two isolated blocking antibody clones against Gal-3BP profoundly inhibited the metastasis of PDAC cells in vivo.
Conclusions

Altogether, our data demonstrates the Gal-3BP is an important therapeutic target in PDAC and proposed its blockade by antibody as a novel, therapeutic option for the inhibition of PDAC metastasis.
SSTP1, A Host Defense Peptide, Exploits the Immunomodulatory Il6 Pathway to Induce Apoptosis in Cancer Cells
Authors: G. Shyla , U. Soumya Krishnan , Mohan Amrutha , Shanmugam Geetha , T.V. Vineethkumar , J. Sreekumar , K.C. Sivakumar , Vasudevan Dileep , Nori Sai Ravi Chandra , Sathi Shijulal Nelson , George Sanil , Maliekal Tessy Thomas
Published: July 2021

Background: The anticancer activities of host defense peptides (HDPs) are mainly attributed to their cell penetrating activity. Accordingly, several approaches based on machine learning, calculating the membrane pore formation ability, have been proposed for the anti-cancer peptide identification. Since the membranolytic activity can lead to nonspecific effects, like hemolysis, the therapeutic application of such molecules is limited. In this context, we considered the immunomodulatory activity of HDPs, which is regulated by specific membrane targets and immunomodulatory pathways in immune cells. As many immunomodulators are aberrantly expressed in cancer cells, the possibility of the activation of an immunomodulatory pathway was investigated for a novel anticancer HDP, SSTP1.Methods: The fourteen mature peptides identified by shotgun cloning from the frog-skin secretions were screened for cytotoxicity in oral cancer cells. The mechanism of action of the selected peptide, SSTP1 was investigated in comparison to its inactive mutant, SSTP2. An RNA-Seq coupled with pathway enrichment analysis was performed to identify the upstream signaling leading to the mitochondrial pathway of apoptosis. Since the activation of the IL6/IL6R pathway was suggested, we performed in silico docking studies to find the binding of SSTP1 to the IL6/IL6Rα/gp130 complex. The dynamic simulation predicted the conformational changes in the active site residues. The confocal co-localization studies, pull-down assay, FRET analysis, western blot and reporter assays were performed to elucidate the role of the IL6/IL6R pathway in SSTP1-induced apoptosis. Specific small molecule inhibitors and neutralizing antibodies were used to ascertain the role of the IL6/IL6Rα/gp130 complex in the activation of JNK/AP1 pathway-dependent cell death.Results: SSTP1, a novel temporin, modulates the IL6 pathway and induces apoptosis when it binds to IL6Rα on the active IL6/IL6Rα/gp130 complex, rearranging the active site residues. In contrast to the IL6 blockers inhibiting JAK/STAT activity, SSTP1 shifts the proliferative IL6/JAK/STAT signaling to the apoptotic IL6/JNK/AP1 pathway. In IL6Rα-overexpressing cancer cells, apoptosis is preferred over the membranolytic activity, upon SSTP1 treatment.Conclusions: Here, we provide the evidence of an HDP-induced signaling through immunomodulators, leading to apoptosis in cancer cells. Our study also implies the importance of identifying the targets of HDPs for their clinical application
Predictors for development of palbociclib-induced neutropenia in breast cancer patients as determined by ordered logistic regression analysis
Authors: Kanbayashi Yuko , Sakaguchi Koichi , Ishikawa Takeshi , Takayama Koichi , Taguchi Tetsuya
Published: July 2021

This retrospective study aimed to identify predictors for the development of palbociclib-induced neutropenia. This study retrospectively analysed 78 breast cancer patients who had received palbociclib at our hospital between January 2018 and May 2020. For the regression analysis of factors associated with palbociclib-induced neutropenia, variables were extracted manually from medical charts. The level of palbociclib-induced neutropenia was evaluated using the National Cancer Institute’s Common Terminology Criteria for Adverse Events (version 5). Multivariate ordered logistic regression analysis was performed to identify predictors for the development of neutropenia. Optimal cut-off thresholds were determined using receiver operating characteristic (ROC) analysis. Values of P < 0.05 (2-tailed) were considered significant. Significant factors identified included concomitant use of statin (odds ratio [OR] = 0.104, 95% confidence interval [CI] = 0.018–0.598; P = 0.011] and body mass index (BMI) (OR = 1.118, 95% CI = 1.007–1.241; P = 0.037). ROC analysis revealed that neutropenia (grade 4) was more likely to occur with a BMI ≥ 22.3 kg/m2. In conclusion, no concomitant use of statins and high BMI were identified as significant predictors for the development of palbociclib-induced neutropenia.