bluuebunny/crossref_metadata_2025 huggingface dataset:
NOTCH Receptors and DLK Proteins Enhance Brown Adipogenesis in Mesenchymal C3H10T1/2 Cells
Authors: Rodríguez-Cano María-Milagros , González-Gómez María-Julia , Sánchez-Solana Beatriz , Monsalve Eva-María , Díaz-Guerra María-José M. , Laborda Jorge , Nueda María-Luisa , Baladrón Victoriano
Published: September 2020

The NOTCH family of receptors and ligands is involved in numerous cell differentiation processes, including adipogenesis. We recently showed that overexpression of each of the four NOTCH receptors in 3T3-L1 preadipocytes enhances adipogenesis and modulates the acquisition of the mature adipocyte phenotype. We also revealed that DLK proteins modulate the adipogenesis of 3T3-L1 preadipocytes and mesenchymal C3H10T1/2 cells in an opposite way, despite their function as non-canonical inhibitory ligands of NOTCH receptors. In this work, we used multipotent C3H10T1/2 cells as an adipogenic model. We used standard adipogenic procedures and analyzed different parameters by using quantitative-polymerase chain reaction (qPCR), quantitative reverse transcription-polymerase chain reaction (qRT-PCR), luciferase, Western blot, and metabolic assays. We revealed that C3H10T1/2 multipotent cells show higher levels of NOTCH receptors expression and activity and lower Dlk gene expression levels than 3T3-L1 preadipocytes. We found that the overexpression of NOTCH receptors enhanced C3H10T1/2 adipogenesis levels, and the overexpression of NOTCH receptors and DLK (DELTA-like homolog) proteins modulated the conversion of cells towards a brown-like adipocyte phenotype. These and our prior results with 3T3-L1 preadipocytes strengthen the idea that, depending on the cellular context, a precise and highly regulated level of global NOTCH signaling is necessary to allow adipogenesis and determine the mature adipocyte phenotype.
Student Feedback to Tailor the CARD™ System for Improving the Immunization Experience at School
Authors: Logeman Charlotte , Taddio Anna , McMurtry C. Meghan , Bucci Lucie , MacDonald Noni , Chalmers Garth , Gudzak Victoria , Shah Vibhuti , Coldham Joanne , Little Cheri , Samborn Tracy , Dribnenki Cindy , Snider Joanne
Published: September 2020

Increasing the comfort of vaccine delivery at school is needed to improve the immunization experience for students. We created the CARD™ (C—Comfort, A—Ask, R—Relax and D—Distract) system to address this clinical care gap. Originally designed for grade 7 students, this study examined the perceptions of grade 9 students of CARD™. Grade 9 students who had experience with school-based immunizations, either as recipients or onlookers (n = 7; 100% females 14 years old) participated. Students answered pre–post surveys, reviewed CARD™ educational materials and participated in a semi-structured focus group discussion. The Consolidated Framework for Implementation Research (CFIR) was used as the framework for analysis of qualitative data. Participants reported positive perceptions of CARD™ educational materials and that CARD™ could fit into the school immunization process. CARD™ improved knowledge about effective coping interventions and was recommended for education of both nurses and students. The results provide preliminary evidence that CARD™ is acceptable and appropriate for implementation in grade 9 school-based immunizations.
Application of Low-Intensity Modified Constraint-Induced Movement Therapy to Improve the Affected Upper Limb Functionality in Infantile Hemiplegia with Moderate Manual Ability: Case Series
Authors: Palomo-Carrión Rocío , Romero-Galisteo Rita-Pilar , Pinero-Pinto Elena , López-Muñoz Purificación , Romay-Barrero Helena , José Francisco García-Muro San
Published: September 2020

Objective: To assess the functionality of the affected upper limb in children diagnosed with hemiplegia aged between 4 and 8 years after applying low-intensity modified Constraint-Induced Movement Therapy (mCIMT). Methods: Prospective case series study. A mCIMT protocol was applied for five weeks, with two hours of containment per day. The study variables were quality of movement of the upper limb, spontaneous use, participation of the affected upper limb in activities of daily living, dynamic joint position, grasp–release action, grasp strength, supination and extension elbow movements. Four measurements were performed, using the quality of upper extremity test (QUEST) scale, the Shriners Hospital for Children Upper Extremity Evaluation (SHUEE) Evaluation, a hand dynamometer and a goniometer. Results: The sample was composed of eight children with moderate manual ability. Statistically significant differences were detected in all the studied variables (p < 0.05) between the pre-treatment and post–treatment results (Week 0–Week 5), except for upper limb dressing, putting on splints and buttoning up. In the first week, the changes were statistically significant, except for protective extension, grasp strength, grasp–release and all functional variables (level of functionality and participation of the patient’s upper limbs) in the SHUEE Evaluation (p > 0.05). The greatest increase occurred in spontaneous use from Assessment 1 to Assessment 4 (p = 0.01), reaching 88.87% active participation in bimanual tasks. The quality of movement of the upper limb exhibited a significant value due to the increase in dissociated movements and grasp (p = 0.01). Conclusion: A low dose (50 h) of mCIMT increased the functionality of children diagnosed with congenital hemiplegia between 4 and 8 years of age with moderate manual ability.
Critical Care Echocardiography as a Routine Procedure for the Detection and Early Treatment of Cardiac Pathologies
Authors: Schmidt Stefan , Dieks Jana-Katharina , Quintel Michael , Moerer Onnen
Published: September 2020

Transthoracic and transesophageal echocardiography are important investigations in the intensive care unit (ICU) to diagnose acute cardiac pathologies and assess the haemodynamic status. Recommendations for critical care echocardiography (CCE) have been published recently, but these still lack an evidence-based foundation. It is not known if performing transthoracic echocardiography (TTE) on a routine basis instead of only when required in acute cases is feasible or clinically useful. In this single-centre prospective observational study, we routinely performed TTE on 111 consecutive non-cardiological, non-cardiothoracic surgical ICU patients in two surgical ICUs in a tertiary care facility. Significant cardiac pathologies were detected in 82 (76.6%) and critical cardiac pathologies in 33 (30.8%) of the 107 patients. The most common critical cardiac pathologies were sPAP > 50 mmHg (19.63%), tricuspid annular plane systolic excursion ≤ 13 mm (9.4%), grade III diastolic dysfunction (8.4%), severe tricuspid valve insufficiency (5.6%) and left ventricular ejection fraction (LV-EF) ˂ 30% (4.7%). Some of the most commonly found cardiac pathologies are not well emphasised in current recommendations and training programs. We observed a progression of the cardiac pathologies previously described in 41 of the patients (91.1%). Patients with echocardiographic abnormalities had a significant survival disadvantage in the ICU. By performing CCE routinely, we observed the range and prevalence of cardiac pathologies that can be detected by echocardiography in critically ill patients. We recommend routine transthoracic CCE in ICU patients for early detection of cardiac pathologies and to help inform early intervention regimens, since cardiac conditions carry a significant survival disadvantage for the ICU patient.
Flexible Real-Time Polymerase Chain Reaction-Based Platforms for Detecting Deafness Mutations in Koreans: A Proposed Guideline for the Etiologic Diagnosis of Auditory Neuropathy Spectrum Disorder
Authors: Lee Sang-Yeon , Oh Doo-Yi , Han Jin Hee , Kim Min Young , Kim Bonggi , Kim Bong Jik , Song Jae-Jin , Koo Ja-Won , Lee Jun Ho , Oh Seung Ha , Choi Byung Yoon
Published: September 2020

Routine application of next-generation sequencing in clinical settings is often limited by time- and cost-prohibitive complex filtering steps. Despite the previously introduced genotyping kit that allows screening of the 11 major recurring variants of sensorineural hearing loss (SNHL) genes in the Korean population, the demand for phenotype- and variant-specific screening kits still remains. Herein, we developed a new real-time PCR-based kit (U-TOP™ HL Genotyping Kit Ver2), comprising six variants from two auditory neuropathy spectrum disorder (ANSD) genes (OTOF and ATP1A3) and five variants from three SNHL genes (MPZL2, COCH, and TMC1), with a distinct auditory phenotype, making this the first genotyping kit dedicated to ANSD. The concordance rate with Sanger sequencing, sensitivity, and specificity of this genotyping kit were all 100%, suggesting reliability. The kit not only allows timely and cost-effective identification of recurring OTOF variants, but it also allows timely detection of cochlear nerve deficiency for those without OTOF variants. Herein, we provide a clinical guideline for an efficient, rapid, and cost-effective etiologic diagnosis of prelingual ANSD. Our study provides a good example of continuing to update new key genetic variants, which will continuously be revealed through NGS, as targets for the newly developed genotyping kit.
Laparoscopic Surgery in COVID-19 Era—Safety and Ethical Issues
Authors: Serban Dragos , Smarandache Catalin Gabriel , Tudor Corneliu , Duta Lucian Nicolae , Dascalu Ana Maria , Aliuș Cătălin
Published: September 2020

(1) Background: The paper aims to review the available evidence regarding the health risk of the aerosolization induced by laparoscopy induced and impact of the COVID-19 pandemic upon minimally invasive surgery. (2) Materials and methods: A systematic review of the literature was performed on PubMed, Medline and Scopus until 10 July. (3) Results: Chemicals, carcinogens and biologically active materials, such as bacteria and viruses, have been isolated in surgical smoke. However, the only evidence of viral transmission through surgical smoke to medical staff is post-laser ablation of HPV-positive genital warts. The reports of SARS-CoV-2 infected patients who underwent laparoscopic surgery revealed the presence of the virus, when tested, in digestive wall and stools in 50% of cases but not in bile or peritoneal fluid. All surgeries did not result in contamination of the personnel, when protective measures were applied, including personal protective equipment (PPE) and filtration of the pneumoperitoneum. There are no comparative studies between classical and laparoscopic surgery. (4) Conclusions: Previously published data showed there is a possible infectious and toxic risk related to surgical smoke but not particularly proven for SARS-CoV-2. Implementing standardized filtration systems for smoke evacuation during laparoscopy, although increases costs, is necessary to increase the safety and it will probably remain a routine also in the future.
Deep Learning Models for Automated Diagnosis of Retinopathy of Prematurity in Preterm Infants
Authors: Huang Yo-Ping , Vadloori Spandana , Chu Hung-Chi , Kang Eugene Yu-Chuan , Wu Wei-Chi , Kusaka Shunji , Fukushima Yoko
Published: September 2020

Retinopathy of prematurity (ROP) is a disease that can cause blindness in premature infants. It is characterized by immature vascular growth of the retinal blood vessels. However, early detection and treatment of ROP can significantly improve the visual acuity of high-risk patients. Thus, early diagnosis of ROP is crucial in preventing visual impairment. However, several patients refrain from treatment owing to the lack of medical expertise in diagnosing the disease; this is especially problematic considering that the number of ROP cases is on the rise. To this end, we applied transfer learning to five deep neural network architectures for identifying ROP in preterm infants. Our results showed that the VGG19 model outperformed the other models in determining whether a preterm infant has ROP, with 96% accuracy, 96.6% sensitivity, and 95.2% specificity. We also classified the severity of the disease; the VGG19 model showed 98.82% accuracy in predicting the severity of the disease with a sensitivity and specificity of 100% and 98.41%, respectively. We performed 5-fold cross-validation on the datasets to validate the reliability of the VGG19 model and found that the VGG19 model exhibited high accuracy in predicting ROP. These findings could help promote the development of computer-aided diagnosis.
Integration of Municipal Air-Conditioning, Power, and Gas Supplies Using an LNG Cold Exergy-Assisted Kalina Cycle System
Authors: Ayou Dereje S. , Eveloy Valerie
Published: September 2020

A Kalina cycle-based integration concept of municipal air-conditioning, electricity and gas is investigated thermodynamically, economically, and environmentally to reduce the carbon intensity of these supplies, with attention to hot climatic conditions. The proposed poly-generation system is driven by low-grade renewable or surplus heat, and utilizes waste exergy from liquefied natural gas vaporization for refrigeration and power augmentation. At nominal conditions (130 °C driving heat), approximately 561 and 151 kJ of refrigeration and useful power per kg of liquefied natural gas regasified are generated by the proposed system, respectively, at effective first-law and exergetic efficiencies of 33% and 35%, respectively. The Kalina sub-system condenser cryogenic heat rejection condition is found to triple the system useful electrical output compared with high ambient temperature condenser heat sinking conditions. Per million ton per annum of liquefied natural gas vaporization capacity, yearly net power savings of approximately 74 GWhe could be achieved compared to standard air-conditioning, electricity, and gas supply systems, resulting in 11.1 kton of natural gas saved and 30.4 kton of carbon dioxide-equivalent emissions avoided annually. The yearly net monetary savings would range from 0.9 to 4.7 million USD per million ton per annum of liquefied natural gas regasified at local subsidized and international electricity market prices, respectively, with corresponding payback periods of 1.7 and 2.5 years, respectively.
Powdery Mildews on Trees and Shrubs in Botanical Gardens, Parks and Urban Green Areas in the Czech Republic
Authors: Mieslerová Barbora , Sedlářová Michaela , Michutová Markéta , Petřeková Veronika , Cook Roger , Lebeda Aleš
Published: September 2020

A total of 103 tree and shrub samples infected with powdery mildew were collected during 2002–2019 from locations within parks, botanical gardens and urban green areas within the Czech Republic and the powdery mildews were morphologically analyzed and identified. The most frequently represented genera were: Erysiphe (27, including former genera Microsphaera and Uncinula), Podosphaera (11, including former genus Sphaerotheca), Phyllactinia (3), Sawadaea (2) and Arthrocladiella (1). New records for the Czech Republic were: E. (U.) arcuata, E. (M.) deutziae, E. (M.) euonymicola, E. (U.) flexuosa, E. (M.) platani, E. (M.) symphoricarpi, E. (M.) vanbruntiana var. sambuci-racemosae, E. (U.) ulmi, Po. amelanchieris, Po. (Sph.) pruinosa and Po. (Sph.) spiraeae. The results were compared with the spectrum of powdery mildew species in the surrounding countries (Slovakia; Hungary; Poland and Germany (Bavaria)).
Anti-Inflammatory and Cytotoxicity Effects of Cudrania tricuspidata Fruits Vinegar in a Co-Culture System with RAW264.7 Macrophages and 3T3-L1 Adipocytes
Authors: Choi Jun-Hui , Park Se-Eun , Yeo Soo-Hwan , Kim Seung
Published: September 2020

Vinegar has been found to have in vitro improvement effect on inflammatory biomarkers, and clinically used to improve inflammation and obesity-related diseases. This study was designed to analyze in vitro anti-inflammatory effects of Cudrania tricuspidata fruits vinegar (CTFV) in a co-culture system with macrophages and adipocytes. We analyzed the physicochemical properties and polyphenolic ingredients of CTFV, and investigated in vitro anti-inflammatory effects of CTFV in a co-culture system with macrophages and adipocytes. The cells were cultured in the presence of CTFV for 24 h in contact with each other, then, harvested. The levels of monocyte chemoattractant protein (MCP)-1, tumour necrosis factor (TNF)-α, inducible nitric oxide synthase (iNOS), nitric oxide (NO), and interleukin (IL)-6 were evaluated by using the Griess reagent, western blot, or enzyme-linked immunosorbent assay assays. We found that increasing levels for NO, iNOS, TNF-α, IL-6 and MCP-1 were caused by LPS treatment and co-culture using the contact method, whereas CTFV efficaciously attenuated inflammatory response by improving inflammatory parameters including NO, iNOS, TNF-α, IL-6 and MCP-1. The present study indicates that CTFV might provide a nutraceutical product or functional food resource for improving inflammation processed via the interaction of adipocytes and macrophages.
The Effect of Algae or Insect Supplementation as Alternative Protein Sources on the Volatile Profile of Chicken Meat
Authors: Gkarane Vasiliki , Ciulu Marco , Altmann Brianne A. , Schmitt Armin O. , Mörlein Daniel
Published: September 2020

The aim of this study was to investigate the differences in the volatile profile of meat from chickens fed with alternative protein diets (such as algae or insect) through two different trials. In Trial 1, broiler chicken at one day of age were randomly allocated to three experimental groups: a basal control diet (C) and two groups in which the soybean meal was replaced at 75% (in the starter phase) and 50% (in the grower phase) with partially defatted Hermetia illucens (HI) larvae or Arthrospira platensis (SP). In Trial 2, broiler chickens were housed and reared similar to Trial 1, with the exception that the experimental diets replaced soybean meal with either 100% partially defatted HI or 100% SP. In both trials, chickens were slaughtered at day 35. Per group, 10 chickens were submitted to volatile analysis by using solid-phase microextraction (HS-SPME) and gas chromatography–mass spectrometry (GC-MS) analysis. Results in both trials showed that levels of several lipid-derived compounds were found to be lower in chickens fed an HI diet, which could be linked to a possibly lower level of polyunsaturated fatty acid content in HI-fed chicken. In addition, the dietary treatments could be discriminated based on the volatile profile, i.e., the substitution of soy with HI or SP distinctively affected the levels of flavor compounds.
Wild Food Plants and Trends in Their Use: From Knowledge and Perceptions to Drivers of Change in West Sumatra, Indonesia
Authors: Pawera Lukas , Khomsan Ali , Zuhud Ervizal A.M. , Hunter Danny , Ickowitz Amy , Polesny Zbynek
Published: September 2020

Wild food plants (WFPs) are often highly nutritious but under-consumed at the same time. This study aimed to document the diversity of WFPs, and assess perceptions, attitudes, and drivers of change in their consumption among Minangkabau and Mandailing women farmers in West Sumatra. We applied a mixed-method approach consisting of interviews with 200 women and focus group discussions with 68 participants. The study documented 106 WFPs (85 species), and Minangkabau were found to steward richer traditional knowledge than Mandailing. Although both communities perceived WFPs positively, consumption has declined over the last generation. The main reasons perceived by respondents were due to the decreased availability of WFPs and changes in lifestyle. The contemporary barriers to consuming WFPs were low availability, time constraints, and a limited knowledge of their nutritional value. The key motivations for their use were that they are free and “unpolluted” natural foods. The main drivers of change were socio-economic factors and changes in agriculture and markets. However, the persistence of a strong culture appears to slow dietary changes. The communities, government and NGOs should work together to optimize the use of this food biodiversity in a sustainable way. This integrated approach could improve nutrition while conserving biological and cultural diversity.
Genetic Hypothesis and Pharmacogenetics Side of Renin-Angiotensin-System in COVID-19
Authors: Gemmati Donato , Tisato Veronica
Published: September 2020

The importance of host genetics and demography in coronavirus disease 2019 (COVID-19) is a crucial aspect of infection, prognosis and associated case fatality rate. Individual genetic landscapes can contribute to understand Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) burden and can give information on how to fight virus spreading and the associated severe acute respiratory distress syndrome (ARDS). The spread and pathogenicity of the virus have become pandemic on specific geographic areas and ethnicities. Interestingly, SARS-CoV-2 firstly emerged in East Asia and next in Europe, where it has caused higher morbidity and mortality. This is a peculiar feature of SARS-CoV-2, different from past global viral infections (i.e., SARS-1 or MERS); it shares with the previous pandemics strong age- and sex-dependent gaps in the disease outcome. The observation that the severest COVID-19 patients are more likely to have a history of hypertension, diabetes and/or cardiovascular disease and receive Renin-Angiotensin-System (RAS) inhibitor treatment raised the hypothesis that RAS-unbalancing may have a crucial role. Accordingly, we recently published a genetic hypothesis on the role of RAS-pathway genes (ACE1, rs4646994, rs1799752, rs4340, rs13447447; and ACE2, rs2285666, rs1978124, rs714205) and ABO-locus (rs495828, rs8176746) in COVID-19 prognosis, suspecting inherited genetic predispositions to be predictive of COVID-19 severity. In addition, recently, Genome-Wide Association Studies (GWAS) found COVID-19-association signals at locus 3p21.31 (rs11385942) comprising the solute carrier SLC6A20 (Na+ and Cl- coupled transporter family) and at locus 9q34.2 (rs657152) coincident with ABO-blood group (rs8176747, rs41302905, rs8176719), and interestingly, both loci are associated to RAS-pathway. Finally, ACE1 and ACE2 haplotypes seem to provide plausible explanations for why SARS-CoV-2 have affected more heavily some ethnic groups, namely people with European ancestry, than Asians.
An Overview of Duplicated Gene Detection Methods: Why the Duplication Mechanism Has to Be Accounted for in Their Choice
Authors: Lallemand Tanguy , Leduc Martin , Landès Claudine , Rizzon Carène , Lerat Emmanuelle
Published: September 2020

Gene duplication is an important evolutionary mechanism allowing to provide new genetic material and thus opportunities to acquire new gene functions for an organism, with major implications such as speciation events. Various processes are known to allow a gene to be duplicated and different models explain how duplicated genes can be maintained in genomes. Due to their particular importance, the identification of duplicated genes is essential when studying genome evolution but it can still be a challenge due to the various fates duplicated genes can encounter. In this review, we first describe the evolutionary processes allowing the formation of duplicated genes but also describe the various bioinformatic approaches that can be used to identify them in genome sequences. Indeed, these bioinformatic approaches differ according to the underlying duplication mechanism. Hence, understanding the specificity of the duplicated genes of interest is a great asset for tool selection and should be taken into account when exploring a biological question.
Prevalence Of, and Factors Associated with, Obesity among the Oldest Old. A Study Protocol for a Systematic Review
Authors: Hajek André , Kretzler Benedikt , König Hans-Helmut
Published: September 2020

Some empirical studies have identified the prevalence of, and factors associated with, obesity among the oldest old. However, there is a lack of a systematic review synthesizing the existing evidence. Therefore, the purpose of our upcoming systematic review is to provide an overview of the evidence provided by observational studies. The current paper presents the protocol for this systematic review. We will search four electronic databases (Medline, PsycINFO, CINAHL, and Cochrane Library). Furthermore, we will perform a manual search (searching reference lists of included studies). Cross-sectional and longitudinal observational studies identifying the prevalence and preferably the factors associated with obesity among the oldest old (80 years and over) will be included. Data extraction will concentrate on study design, assessment of obesity and its associated factors, statistical analysis, sample characteristics, and key findings. We will evaluate the quality of the included studies. Two individuals will perform study selection, data extraction, and evaluation of study quality. We will present the results in figures, summary tables and narrative summaries. If data permits, a meta-analysis will be conducted.
Compositional Nutrient Diagnosis (CND) Applied to Grapevines Grown in Subtropical Climate Region
Authors: Rozane Danilo Eduardo , Vahl de Paula Betania , Wellington Bastos de Melo George , Haitzmann dos Santos Eduardo Maciel , Trentin Edicarla , Marchezan Carina , Stefanello da Silva Lincon Oliveira , Tassinari Adriele , Dotto Lucas , Nunes de Oliveira Filipe , Natale William , Baldi Elena , Toselli Moreno , Brunetto Gustavo
Published: September 2020

Soil analysis is used to estimate nutrient availability, but nutrient concentrations are not always related to yield in most fruit plants, including grapevines. Thus, additional multivariate mathematical models, such as the compositional nutrient diagnosis (CND), which takes into account leaves nutrient concentration, and yield, can contribute to estimating critical levels or sufficiency bands of elements, as well as to detect deficiency and/or excess of nutrients. The aim of the present study was to establish CND standards, and the critical level and sufficiency band of nutrients, in the grapevine Vitis vinifera L., grown in a subtropical climate region. Leaves were collected in 81 vineyards in the Campanha Gaúcha do Rio Grande do Sul region, Southern Brazil, and analyzed for macro- and micro-nutrient concentration. The yield of each vineyard was assessed. Grapevine nutritional status was calculated through the CND method. CND-r2 indices were effective in establishing the nutritional status of grapevines for macro- and micro-nutrients as sub-optimal, excessive, or balanced. The CND methodology established the critical level and sufficiency bands of nutrients more accurately than the current recommendations for grapevines. Multi-nutrient associations were more effective than the single nutrient determination in defining the threshold of a given nutrient that can reduce grapevine yield.
Therapeutic Relationship in eHealth—A Pilot Study of Similarities and Differences between the Online Program Priovi and Therapists Treating Borderline Personality Disorder
Authors: Köhne Sandra , Schweiger Ulrich , Jacob Gitta A. , Braakmann Diana , Klein Jan Philipp , Borgwardt Stefan , Assmann Nele , Rogg Mirco , Schaich Anja , Faßbinder Eva
Published: September 2020

eHealth programs have been found to be effective in treating many psychological conditions. Regarding Borderline Personality Disorder (BPD), few programs have been tested; nevertheless, results are promising. The therapeutic alliance is an important factor predicting treatment outcome in BPD. However, we do not know yet to what extent BPD patients form a therapeutic alliance with an eHealth tool and how this relationship differs from the relationship with their human therapist. This study aims to address this question using priovi, an interactive schema therapy-based eHealth tool for BPD. Semi-structured interviews were conducted to explore how patients perceived the therapeutic alliance with priovi and its differences compared to the alliance with their human therapist (N = 9). Interview data were analyzed following the procedures of qualitative content analysis. Additionally, the Working Alliance Inventory (WAI-SR) was administered in two versions (regarding the human therapist and priovi, N = 16) every three months during the treatment phase of one year. Results indicate that patients were able to form a good therapeutic relationship with priovi, but it differed from the relationship to their human therapist. Important categories were “priovi is helpful, supportive and always there” and “priovi is less flexible”. WAI ratings for the task subscale were high in both relationships but significantly higher in WAItherapist compared to WAIpriovi in two measurements (nine-months measurement: t = 2.76, df = 15, p = 0.015; twelve-months measurement: t = 3.44, df = 15, p = 0.004). These results indicate that BPD patients can form a functioning alliance with an eHealth program and that eHealth programs may be especially useful for psychoeducation and cognitive exercises.
Urinary Acrylonitrile Metabolite Concentrations Before and after Smoked, Vaporized, and Oral Cannabis in Frequent and Occasional Cannabis Users
Authors: Ashley David L. , De Jesús Víctor R. , Abulseoud Osama A. , Huestis Marilyn A. , Milan Daniel F. , Blount Benjamin C.
Published: September 2020

Cannabis use through smoking, vaping, or ingestion is increasing, but only limited studies have investigated the resulting exposure to harmful chemicals. N-acetyl-S-(2-cyanoethyl)-L-cysteine (2CYEMA), a urinary metabolite of acrylonitrile, a possible carcinogen, is elevated in the urine of past-30-day cannabis users compared to non-cannabis users. Five frequent and five occasional cannabis users smoked and vaped cannabis on separate days; one also consumed cannabis orally. Urine samples were collected before and up to 72 h post dose and urinary 2CYEMA was quantified. We compared 2CYEMA pre-exposure levels, maximum concentration, time at maximum concentration for occasional versus frequent users following different exposure routes, and measured half-life of elimination. Smoking cannabis joints rapidly (within 10 min) increased 2CYEMA in the urine of occasional cannabis users, but not in frequent users. Urine 2CYEMA did not consistently increase following vaping or ingestion in either study group. Cigarette smokers had high pre-exposure concentrations of 2CYEMA. Following cannabis smoking, the half-lives of 2CYEMA ranged from 2.5 to 9.0 h. 2CYEMA is an effective biomarker of cannabis smoke exposure, including smoke from a single cannabis joint, however, not from vaping or when consumed orally. When using 2CYEMA to evaluate exposure in cannabis users, investigators should collect the details about tobacco smoking, route of consumption, and time since last use as possible covariates.
The Physical Characteristics of Elite Female Rugby Union Players
Authors: Posthumus Logan , Macgregor Campbell , Winwood Paul , Tout Jamie , Morton Lillian , Driller Matthew , Gill Nicholas
Published: September 2020

This study explored the anthropometric and body composition characteristics of elite female rugby union players, comparing between and within different playing positions. Thirty elite female rugby union players (25.6 ± 4.3 y, 171.3 ± 7.7 cm, 83.5 ± 13.9 kg) from New Zealand participated in this study. Physical characteristics were assessed using anthropometric (height, body mass, skinfolds) and body composition (dual-energy X-ray absorptiometry) measures. Forwards were significantly taller (p < 0.01; d = 1.34), heavier (p < 0.01; d = 2.19), and possessed greater skinfolds (p < 0.01; d = 1.02) than backs. Forwards also possessed significantly greater total (p < 0.01; d = 1.83–2.25) and regional (p < 0.01; d = 1.50–2.50) body composition measures compared to backs. Healthy bone mineral density values were observed in both forwards and backs, with significantly greater values observed at the arm (p < 0.01; d = 0.92) and femoral neck (p = 0.04; d = 0.77) sites for forwards. Tight-five players were significantly heavier (p = 0.02; d = 1.41) and possessed significantly greater skinfolds (p < 0.01; d = 0.97) than loose-forwards. Tight-five also possessed significantly greater total body composition measures (p < 0.05; d = 0.97–1.77) and significantly greater trunk lean mass (p = 0.04; d = 1.14), trunk fat mass (p < 0.01; d = 1.84), and arm fat mass (p = 0.02; d = 1.35) compared to loose-forwards. Specific programming and monitoring for forwards and backs, particularly within forward positional groups, appear important due to such physical characteristic differences.
Adipose Tissue from Lean and Obese Mice Induces a Mesenchymal to Epithelial Transition-Like Effect in Triple Negative Breast Cancers Cells Grown in 3-Dimensional Culture
Authors: Asante Emmanuel C. , Pallegar Nikitha K. , Hoffmann Alica J. , Viloria-Petit Alicia M. , Christian Sherri L.
Published: September 2020

Breast cancer is the second leading cause of cancer-related mortality among women globally with obesity being one risk factor. Obese breast cancer patients have at least a 30% increased risk of death from breast cancer compared to non-obese breast cancer patients because they present with larger tumors and generally have increased rates of metastasis. Moreover, obese breast cancer patients respond more poorly to treatment compared to non-obese patients, particularly pre-menopausal women diagnosed with triple negative breast cancer (TNBC). To help understand the molecular mechanisms underlying the increased metastasis associated with obesity, we previously established a three-dimensional culture system that permits the co-culture of adipocytes and TNBC cells in a manner that mimics an in vivo milieu. Using this system, we demonstrate that white adipose tissue from both lean and obese mice can induce a partial mesenchymal-to-epithelial transition (MET). Triple negative breast cancer cells adopt an epithelial morphology and have an increased expression of some epithelial markers, but they maintain the expression of mesenchymal markers, furnishing the breast cancer cells with hybrid properties that are associated with more aggressive tumors. Thus, these data suggest that adipose tissue has the potential to promote secondary tumor formation in lean and obese women. Further work is needed to determine if targeting the partial MET induced by adipose tissue could reduce metastasis.
Novel Insights into the Biochemical Mechanism of CK1ε and its Functional Interplay with DDX3X
Authors: Bono Bartolo , Franco Giulia , Riva Valentina , Garbelli Anna , Maga Giovanni
Published: September 2020

Casein Kinase 1 epsilon (CK1ε) is a member of the serine (Ser)/threonine (Thr) CK1 family, known to have crucial roles in several biological scenarios and, ever more frequently, in pathological contexts, such as cancer. Recently, the human DEAD-box RNA helicase 3 X-linked (DDX3X), involved in cancer proliferation and viral infections, has been identified as one of CK1ε substrates and its positive regulator in the Wnt/β-catenin network. However, the way by which these two proteins influence each other has not been fully clarified. In order to further investigate their interplay, we defined the kinetic parameters of CK1ε towards its substrates: ATP, casein, Dvl2 and DDX3X. CK1ε affinity for ATP depends on the nature of the substrate: increasing of casein concentrations led to an increase of KmATP, while increasing DDX3X reduced it. In literature, DDX3X is described to act as an allosteric activator of CK1ε. However, when we performed kinase reactions combining DDX3X and casein, we did not find a positive effect of DDX3X on casein phosphorylation by CK1ε, while both substrates were phosphorylated in a competitive manner. Moreover, CK1ε positively stimulates DDX3X ATPase activity. Our data provide a more detailed kinetic characterization on the functional interplay of these two proteins.
The Role of Thiocyanate in Modulating Myeloperoxidase Activity during Disease
Authors: San Gabriel Patrick , Liu Yuyang , Schroder Angie , Zoellner Hans , Chami Belal
Published: September 2020

Thiocyanate (SCN−) is a pseudohalide anion omnipresent across mammals and is particularly concentrated in secretions within the oral cavity, digestive tract and airway. Thiocyanate can outcompete chlorine anions and other halides (F−, Br−, I−) as substrates for myeloperoxidase by undergoing two-electron oxidation with hydrogen peroxide. This forms their respective hypohalous acids (HOX where X− = halides) and in the case of thiocyanate, hypothiocyanous acid (HOSCN), which is also a bactericidal oxidative species involved in the regulation of commensal and pathogenic microflora. Disease may dysregulate redox processes and cause imbalances in the oxidative profile, where typically favoured oxidative species, such as hypochlorous acid (HOCl), result in an overabundance of chlorinated protein residues. As such, the pharmacological capacity of thiocyanate has been recently investigated for its ability to modulate myeloperoxidase activity for HOSCN, a less potent species relative to HOCl, although outcomes vary significantly across different disease models. To date, most studies have focused on therapeutic effects in respiratory and cardiovascular animal models. However, we note other conditions such as rheumatic arthritis where SCN− administration may worsen patient outcomes. Here, we discuss the pathophysiological role of SCN− in diseases where MPO is implicated.
Pathways of Gastric Carcinogenesis, Helicobacter pylori Virulence and Interactions with Antioxidant Systems, Vitamin C and Phytochemicals
Authors: Toh James W. T. , Wilson Robert B.
Published: September 2020

Helicobacter pylori is a class one carcinogen which causes chronic atrophic gastritis, gastric intestinal metaplasia, dysplasia and adenocarcinoma. The mechanisms by which H. pylori interacts with other risk and protective factors, particularly vitamin C in gastric carcinogenesis are complex. Gastric carcinogenesis includes metabolic, environmental, epigenetic, genomic, infective, inflammatory and oncogenic pathways. The molecular classification of gastric cancer subtypes has revolutionized the understanding of gastric carcinogenesis. This includes the tumour microenvironment, germline mutations, and the role of Helicobacter pylori bacteria, Epstein Barr virus and epigenetics in somatic mutations. There is evidence that ascorbic acid, phytochemicals and endogenous antioxidant systems can modify the risk of gastric cancer. Gastric juice ascorbate levels depend on dietary intake of ascorbic acid but can also be decreased by H. pylori infection, H. pylori CagA secretion, tobacco smoking, achlorhydria and chronic atrophic gastritis. Ascorbic acid may be protective against gastric cancer by its antioxidant effect in gastric cytoprotection, regenerating active vitamin E and glutathione, inhibiting endogenous N-nitrosation, reducing toxic effects of ingested nitrosodimethylamines and heterocyclic amines, and preventing H. pylori infection. The effectiveness of such cytoprotection is related to H. pylori strain virulence, particularly CagA expression. The role of vitamin C in epigenetic reprogramming in gastric cancer is still evolving. Other factors in conjunction with vitamin C also play a role in gastric carcinogenesis. Eradication of H. pylori may lead to recovery of vitamin C secretion by gastric epithelium and enable regression of premalignant gastric lesions, thereby interrupting the Correa cascade of gastric carcinogenesis.
Adipose-Derived Mesenchymal Stem Cells do not Affect the Invasion and Migration Potential of Oral Squamous Carcinoma Cells
Authors: Sinha Snehadri , Narjus-Sterba Matilda , Tuomainen Katja , Kaur Sippy , Seppänen-Kaijansinkko Riitta , Salo Tuula , Mannerström Bettina , Al-Samadi Ahmed
Published: September 2020

Mesenchymal stem cells (MSCs) are commonly isolated from bone marrow and adipose tissue. Depending on the tissue of origin, MSCs have different characteristics and physiological effects. In various cancer studies, MSCs have been found to have either tumor-promoting or tumor-inhibiting action. This study investigated the effect of adipose tissue-MSCs (AT-MSCs) and bone marrow-MSCs (BM-MSCs) on global long interspersed nuclear element-1 (LINE-1) methylation, the expression level of microenvironment remodeling genes and cell proliferation, migration and invasion of oral tongue squamous cell carcinoma (OTSCC). Additionally, we studied the effect of human tongue squamous carcinoma (HSC-3)-conditioned media on LINE-1 methylation and the expression of microenvironment remodeling genes in AT-MSCs and BM-MSCs. Conditioned media from HSC-3 or MSCs did not affect LINE-1 methylation level in either cancer cells or MSCs, respectively. In HSC-3 cells, no effect of MSCs-conditioned media was detected on the expression of ICAM1, ITGA3 or MMP1. On the other hand, HSC-3-conditioned media upregulated ICAM1 and MMP1 expression in both types of MSCs. Co-cultures of AT-MSCs with HSC-3 did not induce proliferation, migration or invasion of the cancer cells. In conclusion, AT-MSCs, unlike BM-MSCs, seem not to participate in oral cancer progression.
Neuroprotection by Neurotropin through Crosstalk of Neurotrophic and Innate Immune Receptors in PC12 Cells
Authors: Fukuda Yu , Nakajima Kazuki , Mutoh Tatsuro
Published: September 2020

Infected or damaged tissues release multiple “alert” molecules such as alarmins and damage-associated molecular patterns (DAMPs) that are recognized by innate immune receptors, and induce tissue inflammation, regeneration, and repair. Recently, an extract from inflamed rabbit skin inoculated with vaccinia virus (Neurotropin®, NTP) was found to induce infarct tolerance in mice receiving permanent ischemic attack to the middle cerebral artery. Likewise, we report herein that NTP prevented the neurite retraction in PC12 cells by nerve growth factor (NGF) deprivation. This effect was accompanied by interaction of Fyn with high-affinity NGF receptor TrkA. Sucrose density gradient subcellular fractionation of NTP-treated cells showed heretofore unidentified membrane fractions with a high-buoyant density containing Trk, B subunit of cholera toxin-bound ganglioside, flotillin-1 and Fyn. Additionally, these new membrane fractions also contained Toll-like receptor 4 (TLR4). Inhibition of TLR4 function by TAK-242 prevented the formation of these unidentified membrane fractions and suppressed neuroprotection by NTP. These observations indicate that NTP controls TrkA-mediated signaling through the formation of clusters of new membrane microdomains, thus providing a platform for crosstalk between neurotrophic and innate immune receptors. Neuroprotective mechanisms through the interaction with innate immune systems may provide novel mechanism for neuroprotection.
The Lactate Receptor HCA1 Is Present in the Choroid Plexus, the Tela Choroidea, and the Neuroepithelial Lining of the Dorsal Part of the Third Ventricle
Authors: Hadzic Alena , Nguyen Teresa D. , Hosoyamada Makoto , Tomioka Naoko H. , Bergersen Linda H. , Storm-Mathisen Jon , Morland Cecilie
Published: September 2020

The volume, composition, and movement of the cerebrospinal fluid (CSF) are important for brain physiology, pathology, and diagnostics. Nevertheless, few studies have focused on the main structure that produces CSF, the choroid plexus (CP). Due to the presence of monocarboxylate transporters (MCTs) in the CP, changes in blood and brain lactate levels are reflected in the CSF. A lactate receptor, the hydroxycarboxylic acid receptor 1 (HCA1), is present in the brain, but whether it is located in the CP or in other periventricular structures has not been studied. Here, we investigated the distribution of HCA1 in the cerebral ventricular system using monomeric red fluorescent protein (mRFP)-HCA1 reporter mice. The reporter signal was only detected in the dorsal part of the third ventricle, where strong mRFP-HCA1 labeling was present in cells of the CP, the tela choroidea, and the neuroepithelial ventricular lining. Co-labeling experiments identified these cells as fibroblasts (in the CP, the tela choroidea, and the ventricle lining) and ependymal cells (in the tela choroidea and the ventricle lining). Our data suggest that the HCA1-containing fibroblasts and ependymal cells have the ability to respond to alterations in CSF lactate in body–brain signaling, but also as a sign of neuropathology (e.g., stroke and Alzheimer’s disease biomarker).
A Review of Exosomal Isolation Methods: Is Size Exclusion Chromatography the Best Option?
Authors: Sidhom Karim , Obi Patience O. , Saleem Ayesha
Published: September 2020

Extracellular vesicles (EVs) are membranous vesicles secreted by both prokaryotic and eukaryotic cells and play a vital role in intercellular communication. EVs are classified into several subtypes based on their origin, physical characteristics, and biomolecular makeup. Exosomes, a subtype of EVs, are released by the fusion of multivesicular bodies (MVB) with the plasma membrane of the cell. Several methods have been described in literature to isolate exosomes from biofluids including blood, urine, milk, and cell culture media, among others. While differential ultracentrifugation (dUC) has been widely used to isolate exosomes, other techniques including ultrafiltration, precipitating agents such as poly-ethylene glycol (PEG), immunoaffinity capture, microfluidics, and size-exclusion chromatography (SEC) have emerged as credible alternatives with pros and cons associated with each. In this review, we provide a summary of commonly used exosomal isolation techniques with a focus on SEC as an ideal methodology. We evaluate the efficacy of SEC to isolate exosomes from an array of biological fluids, with a particular focus on its application to adipose tissue-derived exosomes. We argue that exosomes isolated via SEC are relatively pure and functional, and that this methodology is reproducible, scalable, inexpensive, and does not require specialized equipment or user expertise. However, it must be noted that while SEC is a good candidate method to isolate exosomes, direct comparative studies are required to support this conclusion.
Tolerance of Stored Boar Spermatozoa to Autologous Seminal Plasma: A Proteomic and Lipidomic Approach
Authors: Höfner Lisa , Luther Anne-Marie , Palladini Alessandra , Fröhlich Thomas , Waberski Dagmar
Published: September 2020

Long-term exposure of liquid preserved boar spermatozoa to seminal plasma (SP) can cause dramatic sperm injury. This study examined whether boar specificity exists in the sensitivity of spermatozoa to SP and whether correspondent biomarkers can be identified. Consecutive ejaculates (n = 4–5) collected from 19 boars were centrifuged, diluted with a pH-stablising extender with 10% (v/v) autologous SP and evaluated by computer-assisted semen analysis and flow cytometry. Up until 144 h storage, four boars showed consistently high sperm motility, viability and mitochondria activity, and one boar showed consistently low values. Intra-boar variability was high in the other boars. Screening of SP (n = 12 samples) for protein markers using mass spectrometry identified three protein candidates of which the granulin precursor, legumain and AWN were 0.5 to 0.9 log2-fold less abundant (p < 0.05) in SP-resistant compared to SP-sensitive samples. Lipidome analysis by mass spectrometry revealed 568 lipids showing no difference between the SP-groups. The most abundant lipids were cholesterol (42,442 pmol), followed by phosphatidylserine (20,956 pmol) and ether-linked phosphatidylethanolamine (13,039 pmol). In conclusion, three candidate proteins were identified which might be indicative of SP-tolerance of sperm during long-term storage. Noteworthy, a first lipidomic profile of boar SP is presented.