bluuebunny/crossref_metadata_2025 huggingface dataset:
Monocyte Platelet Aggregates in Children with Kawasaki Disease- a Preliminary Study from a Tertiary Care Centre in North-West India
Authors: Vignesh Pandiarajan , Rawat Amit , Shandilya Jitendra Kumar , Sachdeva Man Updesh Singh , Ahluwalia Jasmina , Singh Surjit
Published: September 2020

Background: Platelet activation is an integral part of pathogenesis of Kawasaki disease (KD). However, there is paucity of literature on flow-cytometry based assessment of platelet activation in KD. We aimed to analyse monocyte-platelet aggregates (MPAs), one of the sensitive markers for platelet activation, by flow cytometry in children with KD.Findings: In this single-centre prospective study, we have enrolled 14 children with KD and results were compared with age-matched febrile (n=15) and healthy (n=13) controls. After gating monocytes in side-scatter plot, MPAs were identified based on CD14 and CD41 expression. Two (2) ml of blood samples for children with KD were collected at 3 phases of illness- acute stage before start of intravenous immunoglobulin or aspirin, 24 hours after completion of IVIg infusion, and 3 months after acute episode of KD. Children with KD had a significantly higher MPA% values [Median (IQR)- 41.3% (26.6, 52.7)] when compared with febrile [Median (IQR)- 5.98% (2.98-9.72)] and normal [Median (IQR)- 4.48% (2.57-5.59)] controls, p<0.01. On follow-up, the MPA% showed a gradual decline in children with KD, but even at 3 months, the value [Median (IQR)- 7.55% (4.15-14.6)] was higher compared to healthy controls [Median (IQR)- 4.48% (2.57-5.59)].Conclusions: Our results suggest that MPA% was significantly elevated in acute stages in children with KD and activated platelets may continue to persist even after systemic inflammation has subsided. Future studies are warranted whether objective evidence of platelet activation may guide the use of immunomodulatory and anti-platelet therapy in KD.
Role of Liver Enzymes in Patients with Blunt Abdominal Trauma to Diagnose Liver Injury
Authors: Shrestha Anup , Neupane Harish Chandra , Tamrakar Kishor Kumar , Bhattarai Abhishek , Katwal Gaurav
Published: September 2020

Background: The liver is the second most injured organ following blunt abdominal trauma (BAT) after spleen. Although the computed tomography (CT) scan is considered as the gold standard for diagnosing liver injury in BAT, it is not readily available in every hospital. This study was performed to evaluate the role of aspartate transaminase (AST) and alanine transaminase (ALT) in patients with BAT and its significance in predicting the diagnosis and severity of the liver injury. Method: The study was conducted in Chitwan Medical College Teaching Hospital (CMCTH) from February 2019 to May 2020. During that period 96 patients with BAT presented to the emergency department(ED) of CMCTH. Results: Among the 96 patients admitted with BAT, 38 patients had liver injury and 58 patients had no liver injury. The median length of the intensive care unit (ICU) stay of patients with liver injury was higher than without liver injury. There was a significant difference in the median level of AST and ALT (<0.001) between patients with liver injury and no liver injury. The median of AST and ALT of patients requiring blood transfusion was more than that of the patient not requiring blood transfusion(p<0.05). The area under the ROC curve of AST was 0.89(95% Confidence Interval 0.86-0.98) and of ALT was 0.92(95% Confidence Interval 0.83-0.97). The area under the curve demonstrated that the test was a good predictor for the identification of liver injury and also the severity of liver enzymes. The cut-off values for the liver injury were 106 U/l and 80 U/l for AST and ALT respectively. Based on these values, AST ≥ 106 U/l had a sensitivity of 71.7 %, a specificity of 90 %, a positive predictive value of 86.8 %, and a negative predictive value of 77.6 %. The corresponding values for ALT ≥ 80 U/l were 77.8 %, 94.1%, 92.1% and 82.8 %, respectively. Conclusion: In conclusion, we report the optimal cut-off value of AST and ALT for liver injury in BAT as ≥ 106 U/l and 80 U/l respectively. The elevated level of AST and ALT might assist the emergency physicians and surgeons to timely refer the suspected patients with the liver injury to a tertiary center.
Application of nano-carbon and titanium clip combined labeling in robot-assisted laparoscopic transverse colon cancer surgery
Authors: Lin Nan , Qiu Jiandong , Song Junchuan , Yu Changwei , Fang Yongchao , Wu Weihang , Yang Weijin , Wang Yu
Published: September 2020

Background: Robot-assisted laparoscopic transverse colon tumor surgery requires precise tumor localization. The purpose of this study was to evaluate the safety and efficacy of nano-carbon and titanium clip combination labeling methods in robot-assisted transverse colon tumor surgery.Methods: From January 2018 to January 2019, the clinical data of 16 patients who underwent preoperative nano-carbon and titanium clip combined with robot-assisted laparoscopic transverse colon cancer surgery were retrospectively analyzed.Results: Of the 16 patients, no signs of abdominal pain, fever, or diarrhea were observed after colonoscopy. Two titanium clips were seen on all of the 16 patients' abdominal plain films. Nano-carbon staining sites were observed during the operation, and no staining disappeared or abdominal cavity contamination. All patients underwent R0 resection. The average number of lymph nodes harvsted was 18.23±5.04 (range, 9-32). The average time to locate the lesion under the laparoscopic was 3.03±1.26 min(range, 1-6min), and the average operation time was 321.43±49.23 min (range, 240-400min). All were consistent with the surgical plan, and there was no intraoperative change of surgical procedure or conversion to open surgery.Conclusion: Preoperative colonoscopy combined with nano-carbon and titanium clip is safe and effective in robot-assisted transverse colon cancer surgery. A At the same time, the labeling method shows potential in shortening the operation time, ensuring sufficient safety margin and reducing complications.
Established fibrous peritoneal metastasis in an immunocompetent mouse model similar to clinical immune microenvironment of gastric cancer
Authors: Fujimori Daisuke , Kinoshita Jun , Yamaguchi Takahisa , Nakamura Yusuke , Gunjigake Katsuya , Ohama Takashi , Sato Koichi , Yamamoto Masami , Tsukamoto Tetsuya , Nomura Sachiyo , Ohta Tetsuo , Fushida Sachio
Published: September 2020

Background Peritoneal metastasis (PM) in gastric cancer (GC) is characterized by diffusely infiltrating and proliferating cancer cells accompanied by extensive stromal fibrosis in the peritoneal space. The prognosis of GC with PM is still poor regardless of the various current treatments. In order to elucidate the cause of difficulties in PM treatment, we compared the tumor immune microenvironment (TME) in primary and PM lesions in GC. In addition, a PM model with fibrous stroma was constructed using immunocompetent mice to determine whether its TME was similar to that in patients. Methods Immuno-histochemical analyses of infiltrating immune cells were performed in paired primary and PM lesions from 28 patients with GC. A C57BL/6J mouse model with PM was established using the mouse GC cell line YTN16 either with or without co-inoculation of mouse myofibroblast cell line LmcMF with a-SMA expression. The resected PM from each mouse model was analyzed the immunocompetent cells using immunohistochemistry.Results The number of CD8+ cells was significantly lower in PM lesions than in primary lesions (P<0.01). Conversely, the number of CD163+ cells (M2 macrophages) was significantly higher in PM lesions than in primary lesions (P=0.016). Azan staining revealed that YTN16 and LmcMF co-inoculated tumors were more fibrous than tumor with YTN16 alone (P<0.05). Co-inoculated fibrous tumor also showed an invasive growth pattern and higher progression than tumor with YTN16 alone (P=0.045). Additionally, YTN16 and LmcMF co-inoculated tumors showed lower infiltration of CD8+ cells and higher infiltration of M2 macrophages than tumors with YTN16 alone (P<0.05, P<0.05). These results indicate that LmcMF plays as cancer-associated fibroblasts (CAFs) by crosstalk with YTN16 and CAFs contribute tumor progression, invasion, fibrosis, and immune suppression.Conclusions This model is the first immunocompetent mouse model to accurately reflect the similar to TME of human clinical PM with fibrosis. By using this model, new treatment strategies for PM, such as anti-CAFs therapies, may be developed.
Intelligent prediction of RBC demand in trauma patients using decision tree methods
Authors: Feng Yannan , Xu Zhenhua , Liu Junting , Sun Xiaolin , Wang Deqing , Yu Yang
Published: September 2020

Background: The vital signs of trauma patients are complex and changeable, and the prediction of blood transfusion demand mainly depends on doctors' experience and trauma scoring system; therefore, it cannot be accurately predicted. In this study, a machine learning decision tree algorithm (classification and regression tree, CRT and eXtreme gradient boosting, XGBoost) was proposed for the demand prediction of traumatic blood transfusion to provide technical support for doctors. Methods: A total of 1,371 trauma patients who were diverted to the emergency department from January 2014 to January 2018 were collected from an emergency trauma database. The vital signs, laboratory examination parameters and blood transfusion volume were used as variables, and the non-invasive parameters and all (non-invasive + invasive) parameters were used to construct an intelligent prediction model for RBC demand by logistic regression (LR), CRT and XGBoost. The prediction accuracy of the model was compared with the area under the curve (AUC).Results: The studies we performed showed that non-invasive parameters were used to predict blood transfusion. The LR method was the best, with an AUC of 0.72 (95% confidence interval [CI] 0.657-0.775), which was higher than the CRT AUC of 0.69 (95% CI 0.633-0.751) and the XGBoost AUC of 0.71 (95% CI 0.654-0.756) (P<0.05). The trauma site and shock index are important prediction parameters. For all the prediction parameters, XGBoost was the best, with an AUC of 0.94 (95% CI 0.893-0.981), which was higher than the LR AUC of 0.80 (95% CI 0.744-0.850) and the CRT AUC of 0.82 (95% CI 0.779-0.853) (P<0.05). Haematocrit/Haemoglobin is an important prediction parameter. Conclusions: The classification performance of the intelligent prediction model constructed by the decision tree algorithm is not inferior to that of the traditional LR method. With the increase in the data quantity, the accuracy of the model improved in the iteration process, and the prediction performance continuously improved, which is conducive to clinical application and wide promotion.
The microbiota of the surface, dermis and subcutaneous tissue of dog skin
Authors: García-Fonticoba Rocío , Ferrer Lluis , Francino Olga , Cusco Anna
Published: September 2020

Background. Studies using highly sensitive molecular techniques have detected bacterial communities below the human epidermis. Depending on their abundance and composition, this finding could be clinically relevant. The aim of this study was to determine if bacteria can be detected in the dermis and subcutaneous tissue of dogs without cutaneous disease using two different approaches: traditional cultures and DNA sequencing of the V4 region of bacterial 16S rRNA gene using next-generation sequencing (NGS).Results. Seven healthy dogs were included in the study, and two sets of samples were collected from each subject. Sample sets were composed of a 6-mm abdominal skin biopsy, including epidermis, dermis, and subcutis, a skin surface swab, and an environmental blank sample for contamination control. One set of samples from each dog was submitted for bacterial culture and the other one for bacterial DNA amplification and sequencing. Five different bacterial genera (Staphylococcus, Bacillus, Corynebacterium, Streptococcus, and Enterococcus) were isolated in five out of the seven skin surface swab samples with aerobic microbiological culture methods, while no growth was obtained from the other two samples. Although some DNA could be amplified from epidermal, dermal, and subcutaneous tissue samples, the results of the NGS were similar to those of the blanks.Conclusion. When investigated with aerobic microbiological culture methods, the dermis and subcutaneous tissue of dogs are sterile. NGS techniques lead to the detection of some bacterial DNA, similar to the signal detected in blanks, which does not support the presence of a microbiota in dermis or subcutaneous tissue.
Risk assessment of acute kidney injury following cardiopulmonary bypass
Authors: Wittlinger Thomas , Maus Martin , Kutschka Ingo , Baraki Hassina , Friedrich Martin
Published: September 2020

Background: Acute kidney injury (AKI) is a frequent and serious complication of cardiac surgery, associated with a high incidence of morbidity and mortality. Although the RIFLE criteria serve as a prominent tool to identify patients at high risk of AKI, an optimized diagnosis model in clinical practice is desired.Methods: Based on the SOP-criteria, 365 patients (10%) developed AKI following surgery and were subjected to RRT. In contrast, the incidence of AKI, defined according to the RIFLE criteria, was only 7% (n=251 patients). Prominent risk factors identified by SOP were patients’ sex, valve and combined valve and bypass surgery, deep hypothermia, use of intra-aortic balloon pump (IABP) and previous coronary interventions. Ischemia, reperfusion, blood loss and surgery time also served as significant risk factors for patient evaluated by SOP.Results: Risk assessment by RIFLE differed in as much as most patients with normothermia and those receiving only cardiovascular bypass developed AKI. However, patients’ sex and valve surgery did not serve as a risk factor.Conclusion: Evaluation of patients by the RIFLE versus SOP criteria yielded different results with more AKI patients detected by SOP. Based on the present data, it is concluded that patients may not prone to AKI when surgery and ischemia time will be kept short, when blood loss is mitigated to a minimum and when surgery is performed under non-hypothermic conditions
Extracardiac Imaging Findings in COVID-19 Associated Multisystem Inflammatory Syndrome in Children
Authors: Fenlon Edward Patrick , Chen Susie , Ruzal-Shapiro Carrie , Jaramillo  , Maddocks Alexis B.R.
Published: September 2020

Background – COVID-19 associated Multisystem Inflammatory Syndrome in Children (MIS-C) is an emerging disease that presents with inflammation of multiple organs. The extracardiac radiographic manifestations of this syndrome are not well understood.Objective – We reviewed the radiologic findings of MIS-C in a cohort of children with a confirmed diagnosis of the syndrome.Materials and methods – In a retrospective study from 4/1/2020 to 5/22/2020, we reviewed imaging studies of 38 children with MIS-C, 21 females (55%) and 17 males (45%), with an average age of 9.2 years (range 1.3 – 20 years). Thirty six had chest radiographs, 6 had abdominal radiographs, 12 had abdominal sonograms or MRI, 2 had neck sonograms, and 3 had brain MRI.Results – 28 patients had pulmonary disease, with 24 (86%) having bilateral opacities, mostly diffuse (n=16, 57%). The most common abnormalities were peribronchial thickening (n=19, 68%), ground glass opacities (n=15, 54%) and consolidation (n=5, 18%). Pleural effusions were only present in children 10 years of age and under, and ground glass opacities were seen more often in older patients (59% vs. 26%, P<0.05). On abdominal imaging, small volume ascites was the most common finding (n=6, 50%). Other findings included right lower quadrant bowel wall thickening (n=3, 25%), gallbladder wall thickening (n=3, 25%), and cervical (n=2) or abdominal (n=2) lymphadenophathy. Of 3 patients with brain MRI, one had bilateral parietooccipital abnormalities and another papilledema.Conclusion – COVID-19 associated MIS-C causes a constellation of findings in the chest and abdomen, most often showing bilateral diffuse pulmonary abnormalities and small volume ascites.
Surveillance on schistosomiasis in five provinces of the People’s Republic of China in the post-elimination era
Authors: Guo Jing-yi , Xu Jing , Zhang Lijuan , Shan Lv , Cao Chun-li , Li Shi-zhu , Zhou Xiao-nong
Published: September 2020

Background: The People’s Republic of China (P. R. China) has made significant progress on schistosomiasis control. Among the 12 provinces (municipality, autonomous region) with schistosomiasis endemic in P.R. China, Guangdong, Shanghai, Fujian, Guangxi and Zhejiang provinces (following called as five provinces) had successively eliminated schistosomiasis during 1985-1995. However, given the increasing mobilization of the population and goods, wetland protection, climate changes etc., consolidation of the schistosomiasis elimination in these five provinces remains challenging. In the current study, we sought to understand the epidemic situation in these post-elimination areas and their surveillance capabilities on schistosomiasis. Methods: (Ⅰ) Through the national schistosomiasis reporting system, annual data reflecting the interventions and surveillance on human beings, cattle and snails based on county level from 2005 to 2016 were collected and analyzed to understand the epidemic status of schistosomiasis in five provinces. (Ⅱ) A standardized score sheet was designed to assess the surveillance capacity for schistosomiasis of selected disease control agencies in five provinces and ten counties. Assessment on surveillance capacity consisted of two parts: the first part focused on the capacity of testing skills including schistosomiasis diagnostic skills, identification of snails’ living and infection status; the second part was to assess the knowledge level about schistosomiasis and its control.Results: The comprehensive assessments showed that no local cases in humans and cattle or infected snail were found in these five provinces from 2005, and the surveillance abilities on schistosomiasis of the prevention and control institutions in five provinces were appropriate. However, from 2005 to 2016, a total of 221 imported cases were detected in Zhejiang, Shanghai and Fujian provinces, and 11.98 hm2 of new snail habitats were found in Zhejiang, Shanghai and Guangxi provinces. In addition, snail infestation reoccurred in 247.55 hm2 of former snail habitats since 2011.Conclusions: Elimination of schistosomiasis was consolidated successfully in five provinces of P.R. China due to effective and strong post-elimination surveillance. Comprehensive consolidation strategies should be focused on the elimination of residual snails and the prevention of imported infection sources among the five provinces to consolidate the achievements of schistosomiasis control.
Wilms’ Tumor 1-associating Protein Promotes Prostate Cancer Proliferation via Upregulation of CDK4 Transcript
Authors: Zhang Lei , Gao Yiren , Wang Jun , Liang Linghui , Cheng Yifei , Qi Feng , Wang Zengjun , hua lixin , Cheng Gong
Published: September 2020

Background: Wilms’ tumor 1-associating protein (WTAP) plays an important role in cell physiological function and have attracted increased interest in cancer research recently. Cyclin-dependent protein kinases (CDKs) are known to participate in regulating the cell cycle and often connected to many malignancies in tumor. We aim to explore whether WTAP or CDKs could play an role in the initiation and progression of prostate cancer(PCa) and hope to provide new insights into PCa treatment and prognostic. Methods: Quantitative real-time PCR, western blotting and immunohistochemistry were performed to explore the expression of WTAP and CDK4 in prostate cancer tissues and cell lines. The survival analysis was used to investigate the association between WTAP expression and the clinical outcomes of prostate cancer. Prostate cancer cell lines were stably transfected with lentivirus approach. CCK-8 assay, colony formation assay, cell invasion and migration assay, cell cycle assay and tumorigenesis in nude mice were performed to study the effect of WTAP in prostate cancer cell lines. RNA immunoprecipitation assay, dual-luciferase reporter assay and siRNA transfection were performed to verify the direct binding sites of WTAP with CDK4 transcript.Results: In prostate cancer tissues and cell lines, WTAP was significantly up-regulated and high expression of WTAP was connected to poor clinical outcomes. Additionally, cell function test indicated that overexpression of WTAP in prostate cancer cell lines could promote cell proliferation, while knocking down showed an opposite results. Subcutaneous xenograft tumor model revealed that overexpression of WTAP could induce tumorigenesis in vivo. Mechanism study showed that CDK4 expression could regulate the expression level of WTAP. Moreover, WTAP could directly bind to 3’-UTR of CDK4 transcript and enhance its stability. Furthermore, specific inhibitors of CDK4 as well as small interfering RNA (siRNA) of CDK4 reversed the promotion of proliferation induced by WTAP.Conclusions: These data indicated that WTAP may act as an oncogenic in prostate cancer by directly binding to CDK4 3’-UTR and stabilizing its transcript which might provide new insights into prostate cancer treatment and prognostic.
CD4 T-cell immune stimulation of HER2+ breast cancer cells alters response to trastuzumab in vitro
Authors: Song Patrick , Mansur Amer , Dugger Kari J. , Davis Tessa R. , Howard Grant , Yankeelov Thomas E. , Sorace Anna G.
Published: September 2020

Introduction: The HER2+ tumor immune microenvironment is composed of macrophages, natural killer cells, and tumor infiltrating lymphocytes, which produce pro-inflammatory cytokines. Determining the effect of T-cells on HER2+ cancer cells during therapy could guide immunogenic therapies that trigger antibody-dependent cellular cytotoxicity. This study utilized longitudinal in vitro time-resolved microscopy imaging to measure T-cell influence on trastuzumab in HER2+ breast cancer.Methods: Fluorescently-labeled breast cancer cells (BT474, SKBR3, MDA-MB-453, and MDA-MB-231) were co-cultured with CD4+ T-cells (Jurkat cell line) and longitudinally imaged to quantify cancer cell viability when treated with or without trastuzumab (10, 25, 50 and 100 mg/mL). The presence and timing of T-cell co-culturing was manipulated to determine immune stimulation of trastuzumab-treated HER2+ breast cancer. HER2 and TNF-a expression were evaluated with western blot and ELISA, respectively. Significance was calculated using a two-tailed parametric t-test. Results: The viability of HER2+ cancer cells significantly decreased when exposed to 25 mg/mL trastuzumab and T-cells, compared to cancer cells exposed to trastuzumab without T-cells (p = 0.01). The presence of T-cells significantly increased TNF-a expression in trastuzumab-treated cancer cells (p = 0.02). Conversely, cancer cells treated with TNF-a and trastuzumab had a similar decrease in viability as trastuzumab-treated cancer cells co-cultured with T-cells (p = 0.32).Conclusions: The presence of T-cells significantly increases the efficacy of targeted therapies and suggests trastuzumab may trigger immune mediated cytotoxicity. Increased TNF-a receptor expression suggest cytokines may interact with trastuzumab to create a state of enhanced response to therapy in HER2+ breast cancer, which has potential to reducing tumor burden.
Does Integrated Medical Insurance System (IMIS) Alleviate the Difficulty of Using Cross-Region Health Care for the Migrant Parents in China
Authors: Ma Chao , Huo Shutong , Chen Hao
Published: September 2020

Background

A large number of internal immigrants in the process of urbanization in China is Migrant Parents, the aging group who move to urban area involuntarily to support their family. They are more vulnerable economically and physically than the younger migrants. However, the fragmentation of rural and urban health insurance schemes divided by “hukou” household registration system limit migrant’s access to healthcare services in their resident location. Some provinces have started to consolidate the Urban Resident Basic Medical Insurance and the New Rural Cooperative Medical Scheme as one Integrated Medical Insurance Schemes (IMIS) to reduce the disparity between different schemes and increate the health care utilization of migrants.
Methods

Using China Migrants Dynamic Survey, we used OLS for regression in models.
Results

We found that the migrant parents who are covered by the IMIS are more likely to choose inpatient service and to seek medical treatment in the migrant destination, by improving the convenience of medical expense reimbursement and relieving the economic pressure.
Discussion

The potential mechanisms of our results could be that IMIS alleviates the difficulty of seeking medical care in migrant destination by improving the convenience of medical expense reimbursement and relieving the economic constrain.
Pre-HAART CD4+ T-Lymphocytes as Biomarkers of Post-HAART Immune Recovery in HIV-TB Co-Infected Children
Authors: Gopalakrishnan Vivek , Bose Eliezer , Nair Usha , Cheng Yuwei , Ghebremichael Musie
Published: September 2020

Background Infection with the Human Immunodeficiency Virus (HIV) dramatically increases the risk of developing active tuberculosis (TB). Several studies have indicated that co-infection with TB increases the risk of HIV progression and death. Sub-Saharan Africa bears the brunt of these dual epidemics, with about 2.4 million HIV-infected people living with TB. The main objective of our study was to assess whether the pre-HAART CD4+ T-lymphocyte counts and percentages could serve as biomarkers for post-HAART treatment immune-recovery in HIV-positive children with and without TB co-infection. Methods The data analyzed in this retrospective study were collected from a cohort of 305 HIV-infected children being treated with HAART. A Lehmann family of ROC curves were used to assess the diagnostic performance of pre- HAART treatment CD4+ T-lymphocyte count and percentage as biomarkers for post-HAART immune recovery. The Kaplan–Meier estimator was used to compare differences in post-HAART recovery times between patients with and without TB co-infection.Results We found that the diagnostic performance of both pre-HARRT treatment CD4+ T-lymphocyte count and percentage was comparable and achieved accuracies as high as 74%. Furthermore, the predictive capability of pre-HAART CD4+ T-lymphocyte count and percentage were slightly better in TB-negative patients. Our analyses also indicate that TB-negative patients have a shorter recovery time compared to the TB-positive patients. Conclusions Pre-HAART CD4+ T-lymphocyte count and percentage are stronger predictors of immune recovery in TB-negative pediatric patients, suggesting that TB co-infection complicates the treatment of HIV in this cohort. These findings suggest that the detection and treatment of TB is essential for the effectiveness of HAART in HIV-infected pediatric patients.
Cell wall and cell membrane defects contribute to reduced cell growth at 28 °C in Phaffia rhodozyma mutant strain MK19
Authors: Miao Lili , Chi Shuang , Hou Ting-Ting , Liu Zhi-Pei , Li Ying
Published: September 2020

Background: Phaffia rhodozyma has many desirable properties for astaxanthin production, including rapid heterotrophic metabolism and high cell densities in fermenter culture. The low optimal temperature range (17-21 °C) for cell growth and astaxanthin synthesis in this species presents an obstacle to efficient industrial-scale astaxanthin production. The inhibition mechanism of cell growth at > 21 °C in P. rhodozyma have not been investigated.Results: MK19, a mutant P. rhodozyma strain grows well at moderate temperatures, its cell growth was also inhibited at 28 °C, but such inhibition was mitigated, and low biomass 6 g/L was obtained after 100 h culture. Transcriptome analysis indicated that low biomass at 28 °C resulted from strong suppression of DNA and RNA synthesis in MK19. Growth inhibition at 28 °C was due to cell membrane damage with a characteristic of low mRNA content of fatty acid (f.a.) pathway genes (acc, fas1, fas2), and consequent low f.a. content. Thinning of cell wall and low mannose content (leading to loss of cell wall integrity) also contributed to reduced cell growth at 28 °C in MK19. Levels of astaxanthin and ergosterol, two end-products of isoprenoid biosynthesis (a shunt pathway of f.a. biosynthesis), reached 2000 μg/g and 7500 μg/g respectively; ~2-fold higher than levels at 21 or 25 °C. Abundance of ergosterol, an important cell membrane component, compensated for lack of f.a., making possible the biomass production of 6 g/L for MK19 at 28 °C. Conclusion: Inhibition of growth of P. rhodozyma at 28 °C results from blocking of DNA, RNA, f.a., and cell wall biosynthesis. In MK19, abundant ergosterol made possible biomass production 6 g/L at 28 °C. Significant accumulation of astaxanthin and ergosterol indicated a active MVA pathway in MK19 at 28 °C. Strengthening of the MVA pathway can be a feasible metabolic engineering approach for enhancement of astaxanthin synthesis in P. rhodozyma. The present findings provide useful mechanistic insights regarding adaptation of P. rhodozyma to 28 °C, and improved understanding of feasible metabolic engineering techniques for industrial scale astaxanthin production by this economically important yeast species.
Aptamer targeted therapy potentiates immune checkpoint blockade in triple-negative breast cancer
Authors: Camorani Simona , Passariello Margherita , Agnello Lisa , Esposito Silvia , Collina Francesca , Cantile Monica , Bonito Maurizio Di , Ulasov Ilya V. , Fedele Monica , Zannetti Antonella , De Lorenzo Claudia , Cerchia Laura
Published: September 2020

Background: Triple-negative breast cancer (TNBC) is a uniquely aggressive cancer with high rates of relapse due to resistance to chemotherapy. TNBC expresses higher levels of programmed cell death-ligand 1 (PD-L1) compared to other breast cancers, providing the rationale for the recently approved immunotherapy with anti-PD-L1 monoclonal antibodies (mAbs). A huge effort is dedicated to identify actionable biomarkers allowing for combination therapies with immune-checkpoint blockade. Platelet-derived growth factor receptor β (PDGFRβ) is highly expressed in invasive TNBC, both on tumor cells and tumor microenvironment. We recently proved that tumor growth and lung metastases are impaired in mouse models of human TNBC by a high efficacious PDGFRβ aptamer. Hence, we aimed at investigating the effectiveness of a novel combination treatment with the PDGFRβ aptamer and anti-PD-L1 mAbs in TNBC.Methods: The targeting ability of the anti-human PDGFRβ aptamer toward the murine receptor was verified by streptavidin-biotin assays and confocal microscopy, and its inhibitory function by transwell migration assays. The anti-proliferative effects of the PDGFRβ aptamer/anti-PD-L1 mAbs combination was assessed in human MDA-MB-231 and murine 4T1 TNBC cells, both grown as monolayer or co-cultured with lymphocytes. Tumor cell lysis and cytokines secretion by lymphocytes were analyzed by LDH quantification and ELISA, respectively. Orthotopic 4T1 xenografts in syngeneic mice were used for dissecting the effect of aptamer/mAb combination on tumor growth, metastasis and lymphocytes infiltration. Ex vivo analyses through immunohistochemistry, RT-qPCR and immunoblotting were performed. Results: We show that the PDGFRβ aptamer potentiates the anti-proliferative activity of anti-PD-L1 mAbs on both human and murine TNBC cells, according to its human/mouse cross-reactivity. Further, by binding to activated human and mouse lymphocytes, the aptamer enhances the anti-PD-L1 mAb-induced cytotoxicity of lymphocytes against tumor cells. Importantly, the aptamer heightens the antibody efficacy in inhibiting tumor growth and lung metastases in mice. It acts on both tumor cells, inhibiting Akt and ERK1/2 signaling pathways, and immune populations, increasing intratumoral CD8+T cells and reducing FOXP3+Treg cells. Conclusion: Co-treatment of PDGFRβ aptamer with anti-PD-L1 mAbs is a viable strategy, thus providing for the first an evidence of the efficacy of PDGFRβ/PD-L1 co-targeting combination therapy in TNBC.
Single cell RNA sequencing of infantile hemangioma reveals cellular heterogeneity
Authors: Yu Zhang , Chen Xuanfeng , Cai Ren , Chen Qianyi , Chang Lei , Qiu Yajing , Chen Zongan , Ying Hanru , Hua Cheng , Gao Wei , Chen Hui , Jin Yunbo , Ma Gang , Pan Weijun , Lin Xiaoxi
Published: September 2020

The authors have requested that this preprint be removed from Research Square.
Clinical Utility of Liquid Biopsy for EGFR Driver, T790M Mutation and EGFR Amplification in Plasma in Patients with Aacquired Resistance to Afatinib
Authors: Oya Yuko , Yoshida Tatsuya , Asada Kazuhiro , Oguri Tetsuya , Inui Naoki , Morikawa Sayako , Ito Kentaro , Kimura Tomoki , Kunii Eiji , Matsui Takashi , Kubo Akihito , Kato Tatsuo , Abe Takashi , Tsuda Takeshi , Hida Toyoaki
Published: September 2020

Background: Cell-free DNA (cfDNA) genotyping in plasma using the cobas EGFR Mutation Test v2 (cobas) is the first liquid biopsy as a companion diagnosis to identify the EGFR T790M mutation (T790M) after the failure of treatment of EGFR-tyrosine kinase inhibitor (TKI) (1st generation, gefitinib [G] and erlotinib [E] and afatinib [A]). This study investigated the clinical utility of a liquid biopsy for patients who acquired resistance to afatinib.Methods: We prospectively collected plasma from 51 patients who had acquired resistance to afatinib between April 2015 and November 2016 to evaluate the frequency of T790M using the cobas and digital droplet PCR (UMIN000025112). Additionally, we retrospectively reviewed 38 patients who tested by cobas in plasma after G/E failure to compare for T790M detection between A and with G/E.Results: The detection rate of EGFR sensitive mutation (EGFR sens.) and T790M in plasma in patients treated with A (A group) as an initial EGFR-TKI was lower than with G/E followed by A (G/E→A), although the differences were not significant (EGFR sens.: 41% (A) vs. 67% (G/E→A), P=0.1867; and T790M: 8% (A) vs. 17% (G/E→A), P=0.5798). In first-line setting, the detection rate for EGFR-sens. and T790M in plasma by cobas was lower in A group than in G/E group, although there was no significant difference (EGFR sens.: 34% (A) vs. 52% (G/E), P=0.2072; and T790M: 10% (A) vs. 27% (G/E), P=0.1161).Conclusion: The detection of EGFR sens. and T790M in plasma by cobas might be insufficient in patients treated with afatinib than with G/E.
Dexamethasone Blunts Post-Spinal Hypotension in Geriatric Patients Undergoing Orthopedic Surgery: A Double Blind, Placebo-Controlled Study
Authors: Ashoor Tarek , Hussien Noha S. , Anis Sherif G. , Esmat Ibrahim M.
Published: September 2020

Background. Post-spinal hypotension in elderly is challenging, correction by fluids either colloids or crystalloids or by vasoconstrictors pose the risk of volume overload or compromising cardiac conditions. Dexamethasone is used to treat conditions manifested by decrease of peripheral vascular resistance; we were the first to test the hypothesis of its role in preventing or decreasing the incidence of post-spinal hypotension.Methods. 110 patients aged 60 years or more were recruited in the study, 55 in the Dexamethasone group (D group) were given 8 mg Dexamethasone 2 hours preoperatively, and 55 were given placebo (C group).Variations in blood pressure and heart rate and need of vasoconstrictors and/or atropine following spinal anesthesia (SA) was done. SA was done by subarachnoid injection of 3 ml of hyperbaric bupivacaine.Results. Demographic data and the quality of sensory and motor block were comparable between groups. At 5, 10 minutes; systolic, diastolic and, mean arterial pressures were significantly higher in group D. At 20 minutes readings were non significant between groups. Heart rate changes didn’t show any significance. Need for ephedrine was less in group D, side effects were less in the D group. Conclusion. Dexamethasone may attenuate post-spinal hypotension in elderly, with a favorable response against nausea, vomiting and shivering that associate spinal anesthesia.
Photo-induced Afterglow of Carbon Dots for Dynamic Patterning
Authors: Liu Yanfeng , Huang Xin , Niu Zuoji , Wang Dongni , Gou Huilin , Liao Qiaobo , Xi Kai , An Zhongfu , Jia Xudong
Published: September 2020

Flexible materials with afterglow feature have received considerable attention in the field of flexible electronics. To date, it is still a formidable challenge to develop flexible materials with dynamic long afterglow for practical applications. Herein, we report for the first time photo-induced long afterglow in a flexible solid composite film made of luminescent carbon dots (CDs) and the oxygen-permeable poly pyrrolidone (PVP). The phosphorescent afterglow of the film can be reversibly activated by photo-irradiation and de-activated by thermal treatment. Impressively, the photo-activation leads to a significant luminescence lifetime enhancement with a factor of over 3900, and editable afterglow patterns with fine resolution exceeding 1280 dpi are readily achieved by masking and lithography. Furthermore, the retention time of such memorized optical patterns can be tuned from minutes to days by varying temperature, enabling the unique time-temperature indicating function. These findings not only enrich both the library of the afterglow materials vastly, but also extend the scope of the potiential applications of CDs materials.
COVID-19 Lung CT Image Segmentation Using Deep Learning Methods: UNET Vs. SegNET
Authors: Saood Adnan , hatem iyad
Published: September 2020

Background: Currently, there is an urgent need for efficient tools to assess in the diagnosis of COVID-19 patients. In this paper, we present feasible solutions for detecting and labeling infected tissues on CT lung images of such patients. Two structurally-different deep learning techniques, SegNet and UNET, are investigated for semantically segmenting infected tissue regions in CT lung images. Methods: We propose to use two known deep learning networks, SegNet and UNET, for image tissue classification. SegNet is characterized as scene segmentation network and UNet as a medical segmentation tool. Both networks were exploited as binary segmentors to discriminate between infected and healthy lung tissue, and as multi-class segmentors to learn the infection type on the lung. Each network is trained using 72 data images, validated on 10 images and tested against the left 18 images. Several statistical scores are calculated for the results and tabulated accordingly. Results: The results show the superior ability of SegNet in classifying infected/non-infected tissues compared to the other methods (with 0.95 mean accuracy), while the UNET shows better results as a multi-class segmentor (with 0.91 mean accuracy). Conclusion: Semantically segmenting CT scan images of COVID-19 patients is a crucial goal because it would not only assist in disease diagnosis , but also help in quantifying the severity of the disease ,and hence, prioritize the population treatment accordingly. We propose computer-based techniques that prove to be reliable as detectors for infected tissue in lung CT scans. The availability of such a method in today’s pandemic would help automate, prioritize, fasten, and broaden the treatment of COVID-19 patients globally.
Transcriptomic Analysis Reveals Salt Tolerance Mechanisms Present in Date-Plum Persimmon Rootstock (Diospyros Lotus)
Authors: Gil_Muñoz Francisco , Delhomme Nicolas , Quiñones Ana , Naval Maria del Mar , Badenes Maria , Gil Rosario García
Published: September 2020

Background Drought and salinity are two of the main challenges in agriculture. In many areas, crop production needs solutions to adapt the grown species to the increasing salinity. Research on physiological and molecular responses activated by salinity in plants is needed to elucidate mechanisms of salinity tolerance. Transcriptome profiling (RNA-Seq) is a powerful tool to study the transcriptomic profile of genotypes under stress conditions. In temperate fruit tree species, tree grafting on salinity tolerant rootstocks is a common method to compensate for the cultivar saline sensitivity. Persimmon species have different levels of tolerance to salinity, knowledge of this variability provides the basics for development of salt tolerant rootstocks.Results In this study, we conducted a physiological and transcriptomic profiling of roots and leaves in tolerant and sensitive plants of persimmon rootstock, Diospyros lotus, grown under saline and control conditions. Results from characterization of the physiological responses along with gene expression changes in roots and leaves allowed identifying several salt-tolerance mechanisms related to Ion transport and thermospermine synthesis. Differences were observed in putative H+/ATPases that allow transmembrane ionic transport and Chloride channel protein-like genes. Furthermore, an overexpression of thermospermine synthase found in the roots of tolerant plants may indicate that alterations in root architecture could act as an additional mechanism of response to salt stress. Conclusions Results indicate that D. lotus presents a genetic variability for salt tolerance trait related to the regulation of chloride transport, transmembrane electrochemical potential and thermospermine root synthesis. The study provides knowledge on mechanism of salt stress tolerance in persimmon for further breeding of tolerant persimmon rootstocks.
The Plasma Amyloid Beta 42 (Aβ42) and Proteomics Profile Related to Canine Cognitive Dysfunction Syndrome (CCDS) in Thailand
Authors: Phochantachinda Sataporn , Chantong Boonrat , Reamtong Onrapak , Chatchaisak Duangthip
Published: September 2020

Background: Canine cognitive dysfunction syndrome (CCDS) is a progressive neurodegenerative disorder found in senior dogs. Due to the lack of biological markers, CCDS is commonly underdiagnosed. The aim of this study was to identify potential plasma biomarkers using proteomics techniques and to increase our understanding of the pathogenic mechanism of the disease. Plasma amyloid beta 42 (Aβ42) has been seen to be a controversial biomarker for CCDS. Proteomics analysis was performed for protein identification and quantification.Results: Within CCDS, ageing, and adult dogs, 87 proteins were identified specific to Canis spp. in the plasma samples. Of 87 proteins, 45 and 52 proteins were changed in the ageing and adult groups, respectively. Several distinctly expressed plasma proteins identified in CCDS were involved in complement and coagulation cascades and the apolipoprotein metabolism pathway. Plasma Aβ42 levels considerably overlapped within the CCDS and ageing groups. In the adult group, the Aβ42 level was low compared with that in the other groups. Nevertheless, plasma Aβ42 did not show a correlation with the Canine Cognitive Dysfunction Rating scale (CCDR) score in the CCDS group (p = 0.125, R2 = 0.27).Conclusions: Our present findings suggest that plasma Aβ42 does not show potential for use as a diagnostic biomarker in CCDS. The nano-LC-MS/MS data revealed that the predictive underlying mechanism of CCDS was the co-occurrence of inflammation-mediated acute phase response proteins and complement and coagulation cascades that partly functioned by apolipoproteins and lipid metabolism. Some of the differentially expressed proteins may serve as potential predictor biomarkers along with Aβ42 in plasma for improved CCDS diagnosis.
Bright triplet excitons for efficient X-ray excited luminescence
Authors: Wang Xiao , Shi Huifang , Ma Huili , Ye Wenpeng , Song Lulu , Zan Jie , Yao Xiaokang , Ou Xiangyu , Yang Guohui , Zhao Zhu , Singh Manjeet , Lin Chongyang , Wang He , Jia Wenyong , Wang Qian , Zhi Jiahuan , Dong Chaomin , Jiang Xueyan , Tang Yongan , Xie Xiaoji , Yang Yang , Wang Jianpu , Chen Qiushui , Wang Yu , Yang Huang-Hao , Zhang Guoqing , An Zhongfu , Liu Xiaogang , Huang Wei
Published: September 2020

Materials that exhibit X-ray excited luminescence have great potential in radiation detection, security inspection, biomedical applications, and X-ray astronomy1-4. However, such materials are almost exclusively limited to inorganic crystals, which are typically prepared under high temperatures5. Herein, we report a design principle of purely organic phosphors to boost X-ray excited luminescence with sufficient utilization of triplet excitons. Our experimental data reveal that proportion of emission from bright triplet excitons is significantly improved upon X-ray irradiation, compared with UV excitation. These organic phosphors have a detection limit of 33 nGy/s, which is 167 times lower than the standard dosage for X-ray medical examinations. We further demonstrated their potential application in X-ray radiography, which can be conveniently recorded using a digital camera. These findings illustrate a fundamental principle to design efficient X-ray excited purely organic phosphors, propelling the development of radioluminescence related applications.
Simultaneous Tracking of Capsid, Envelope Protein Localization in Living Cells Infected with Double Fluorescent Duck Enteritis Virus
Authors: Chen Liu , Ni Zheng , Hua Jionggang , Ye Weicheng , Liu Keshu , Yun Tao , Zhu Yinchu , Zhang Cun
Published: September 2020

Background

Duck enteritis virus can cause an acute, contagious and lethal disease of many species of waterfowl. An infectious bacterial artificial chromosome clone of duck enteritis virus (DEV) vaccine strain pDEV-EF1 has been constructed in our previous study. To visually studying DEV, a double-labeled recombinant virus have been constructed.
Methods

based on pDEV-EF1, a recombinant mutated clone pDEV-UL35(c)CFP-gC(c)mRFP which carries a red fluorescent protein(mRFP) gene attached to the viral envelope protein gC, combined with a cyan fluorescent protein (CFP) gene fused to the viral capsid UL35 gene was constructed by two-step Red/ET recombination and the recombinant virus rDEV-UL35(c)CFP-gC(c)mRFP was rescued from chicken embryo fibroblasts (CEFs) by calcium phosphate precipitation. Then protein expression were detected by Western blot analysis, and subcellular location of gC and UL35 were observed by confocal microscopy. Viral morphogenesis were observed by transmission electron microscopy (TEM).
Results

The recombinant virus rDEV-UL35(c)CFP-gC(c)mRFP was rescued from chicken embryo fibroblasts (CEFs) by calcium phosphate precipitation. Western blot analysis showed UL35-CFP and gC-mRFP are both expressed in fusion forms in rDEV-UL35(c)CFP-gC(c)mRFP-infected CEFs, and subcellular location study showed gC-mRFP was mainly localized in whole cell at 36 h.p.i. and 48 h.p.i.; and then mostly migrated to cytoplasm after 60 h.p.i.; UL35-CFP was localized in nucleus in all stages of virus infection. Transmission electron microscopy indicated that viral particles at different stages of morphogenesis (A capsids, B capsids, C capsids) were observed in virus-infected cells. However,mature C capsids was less in rDEV-UL35(c)CFP-gC(c)mRFP-infected cells than rDEV-dEF1GFP and rDEV-gC(c)mRFP-infected samples.
Conclusions

This study has laid a foundation for visually studying localization, transportation of DEV capsid protein and envelope glycoprotein, as well as virus assembly, virion movement and virus-cell interaction.