bluuebunny/crossref_metadata_2025 huggingface dataset:
Optimization of expression, purification and secretion of functional recombinant human growth hormone in Escherichia coli using modified staphylococcal protein a signal peptide
Authors: Rigi Garshasb , Rostami Amin , Ghomi Habib , Ahmadian Gholamreza , Mirbagheri Vasiqe Sadat , Jeiranikhameneh Meisam , Vahed Majid , Rahimi Sahel
Published: August 2021

Background

Human Growth Hormone (hGH) is a glycoprotein released from the pituitary gland. Due to the wide range of effects in humans, any disruption in hGH secretion could have serious consequences. This highlights the clinical importance of hGH production in the treatment of different diseases associated with a deficiency of this hormone. The production of recombinant mature hormone in suitable hosts and secretion of this therapeutic protein into the extracellular space can be considered as one of the best cost-effective approaches not only to obtain the active form of the protein but also endotoxin-free preparation. Since the natural growth hormone signal peptide is of eukaryotic origin and is not detectable by any of the Escherichia coli secretory systems, including Sec and Tat, and is therefore unable to secrete hGH in the prokaryotic systems, designing a new and efficient signal peptide is essential to direct hGh to the extracellular space.

Results

In this study, using a combination of the bioinformatics design and molecular genetics, the protein A signal peptide from Staphylococcus aureus was modified, redesigned and then fused to the mature hGH coding region. The recombinant hGH was then expressed in E. coli and successfully secreted to the medium through the Sec pathway. Secretion of the hGH into the medium was verified using SDS-PAGE and western blot analysis. Recombinant hGH was then expressed in E. coli and successfully secreted into cell culture medium via the Sec pathway. The secretion of hGH into the extracellular medium was confirmed by SDS-PAGE and Western blot analysis. Furthermore, the addition of glycine was shown to improve hGH secretion onto the culture medium. Equations for determining the optimal conditions were also determined. Functional hGH analysis using an ELISA-based method confirmed that the ratio of the active form of secreted hGH to the inactive form in the periplasm is higher than this ratio in the cytoplasm.

Conclusions

Since the native signal protein peptide of S. aureus protein A was not able to deliver hGH to the extracellular space, it was modified using bioinformatics tools and fused to the n-terminal region of hGh to show that the redesigned signal peptide was functional.
Acute arthritis of the right temporomandibular joint due to Lyme disease: a case report and literature review
Authors: Weise Christina , Schulz Matthias C. , Frank Karin , Cetindis Marcel , Koos Bernd , Weise Hannes
Published: August 2021

Background

Lyme disease is the most frequent tick-borne infectious disease in Europe. It often presents with a wide variety of symptoms. For this reason, affection of the temporomandibular joint (TMJ) caused by Lyme disease (LD) can be misdiagnosed as a common temporomandibular disorder (TMD).

Case presentation

The purpose of this case report of a 25-year-old woman presenting to the Departments of Orthodontics and Oral and Maxillofacial Surgery with extensive symptoms of temporomandibular disorder is to illustrate the delayed diagnosis of Lyme disease which was only made after extensive therapy of the temporomandibular joint. The specialist literature only reports a few cases of patients suffering from Lyme disease with TMJ manifestations.

Conclusion

This case report and the relevant literature review aim to emphasize the importance of accurate request of medical history and differential diagnosis of acute TMJ arthritis and arthralgia. Early interdisciplinary diagnosis of Lyme disease and early antibiotic therapy are essential to avoid misdiagnosis and unnecessary, sometimes invasive, therapies.
Work engagement and associated factors among dental nurses in China
Authors: Wang Yujing , Gao Yuqin , Xun Yang
Published: August 2021

Background

Work engagement is affected by many factors. The level of work engagement among dental nurses is unknown.

Methods

A cross-sectional questionnaire survey was conducted among 215 dental nurses. The Utrecht Work Engagement Scale, Chinese Nurse Stressors Scale, Work-related Acceptance and Action Questionnaire, Multi-dimensional Scale of Perceived Social Support, and General Well-Being Schedule were applied to measure Chinese nurses’ work engagement, job stress, psychological flexibility, perceived social support and subjective well-being, respectively. Univariate analysis was used to identify the relationships of work engagement with demographic and psychological characteristics. Hierarchical linear regression analysis was applied to test the variance in work engagement accounted for by factors related to work engagement in the univariate analysis.

Results

The level of work engagement among Chinese dental nurses was moderate or above. Work engagement was positively associated with perceived social support, psychological flexibility and subjective well-being but negatively correlated with job stress. The hierarchical regression analysis showed that age, job stress, psychological flexibility and subjective well-being were significantly correlated with work engagement, though perceived social support was not, all of those psychological variables together explained 34.7% of the variance in work engagement.

Conclusions

Dental nurses in China had an acceptable level of work engagement in terms of vigour, dedication and absorption. Increased job stress resulted in lower work engagement. Nurses who had higher levels of perceived social support, psychological flexibility and subjective well-being also had higher work engagement. It is necessary to understand the job stress of nurses, strengthen nurses’ social support, relieve nurses’ job stress, improve nurses’ psychological flexibility and subjective well-being, which will improve nurses’ work engagement levels.
Associations between mental health problems and risky oral and sexual behaviour in adolescents in a sub-urban community in Southwest Nigeria
Authors: Folayan Morenike Oluwatoyin , Arowolo Olaniyi , Mapayi Boladale , Chukwumah Nneka Maureen , Alade Michael A. , Yassin Randa H. , El Tantawi Maha
Published: August 2021

Background

This study determined the association between mental health and risky oral health and sexual health behaviours.

Methods

A household cross-sectional survey was conducted in Ile-Ife, Nigeria between December 2019 and January 2020. Data were collected from 10 to 19-year-old on the sociodemographic profile (age, sex at birth and socioeconomic status); mental health problems (psychological distress, depressive symptoms and suicidal ideation); and mental (smoking habit, consumption of alcohol, use of psychoactive substances), sexual (history of vaginal or anal sexual intercourse; transactional sex, multiple sex partners, use of condom at last sexual intercourse) and oral (frequency of daily tooth brushing, daily frequency of consumption of refined carbohydrate in-between-meals, frequency of use of dental floss, history of dental service utilization in the last 12 months and dental anxiety) health risk factors. Binary logistic regression analysis was conducted to determine the association between risky oral (neglecting to brush twice daily and frequent consumption of refined carbohydrates in-between-meals), and sexual (neglecting to use condoms during the last sex act and having multiple sex partners) health behaviours as outcome variables, and mental health status as the explanatory variables. An ordinal logistic regression model was also developed where the outcome variable was the number of risky health behaviours. The models were adjusted for the socio-demographic variables and history of dental service utilisation in the last 12 months of the survey.

Results

High psychological distress was significantly associated with lower odds of frequent consumption of refined carbohydrates in-between-meals (AOR = 0.32; 95%CI 0.23, 0.47), and having multiple sex partners (AOR = 0.10; 95%CI 0.02, 0.57); but higher odds of having a higher number of risky behaviours (AOR = 3.04; 95%CI 2.13, 4.33). Having depressive symptoms was significantly associated with higher odds of neglecting to use condom at the last sexual intercourse (AOR = 7.20; 95%CI 1.94, 26.76) and having multiple partners (AOR = 95.43; 95%CI 24.55, 370.90). Suicidal ideation was significantly associated with lower odds of neglecting to use condom at the last sexual intercourse (AOR = 0.00; 95%CI 0.00, 0.00) and having multiple sex partners (AOR = 0.00; 95%CI 0.00, 0.00).

Conclusion

The associations between psychological distress and oral and sexual health risk behaviours in adolescents seem complex and need to be studied further.
Characterizing circulating nucleosomes in the plasma of dogs with lymphoma
Authors: Dolan Christopher , Miller Tasha , Jill Jarvis , Terrell Jason , Kelly Theresa Kathleen , Bygott Thomas , Wilson-Robles Heather
Published: August 2021

Background

Nucleosomes consist of DNA wrapped around a histone octamer core like beads on a string so that DNA can be condensed as chromatin into chromosomes. Diseases such as cancer or inflammation lead to cell death where chromatin is fragmentated and released as mononucleosomes into the blood. The Nu.Q™ H3.1 assay measures total nucleosome concentration in plasma of humans and has been used to detect and identify cancer even at early stages. The objectives of this study were to determine if nucleosome levels could be used to distinguish between healthy dogs and dogs with various stages of lymphoma (LSA) using the Nu.Q™ H3.1 assay.

A total of 126 dogs diagnosed with LSA and 134 healthy controls were recruited for this study. Plasma was collected from each dog and stored in K2-EDTA tubes. The LSA patient samples were recruited from TAMU or purchased from various biobanks. All control cases were recruited from TAMU.

Results

Dogs with LSA had an approximately 7-fold increase in their plasma nucleosome concentrations compared to controls (AUC 87.8%). Nucleosome concentrations increased with cancer stage and dogs with B cell lymphomas had significantly higher nucleosome concentrations than dogs with T cell lymphomas.

Conclusions

The Nu.Q™ H3.1 assay was able to reliably detect elevated nucleosome concentrations in the plasma of dogs with LSA. Furthermore, it appears that nucleosomes are useful for differentiating cancer from healthy individuals in canines.
The risk of consequent nephropathy following initial weight loss in diabetic patients treated with sodium glucose cotransporter 2 inhibitors
Authors: Chan Yi-Hsin , Chen Shao-Wei , Chao Tze-Fan , Kao Yi-Wei , Huang Chien-Ying , Chu Pao-Hsien
Published: August 2021

Background

There is a controversy over the association between obesity and the risk of renal events in patients with type 2 diabetes mellitus (T2DM). Furthermore, whether body weight (BW) loss following sodium glucose cotransporter 2 inhibitor (SGLT2i) treatment associated with risk of adverse renal events is unknown.

Methods

We used medical data from a multi-center healthcare provider in Taiwan, enrolling 8992 T2DM patients with a baseline/following-up BW data available after around 12 weeks of SGLT2i treatment, from June 1, 2016 to December 31, 2018. Patients were followed up until the occurrence of composite renal outcome (estimated glomerular filtration rate decline > 40% or end-stage kidney disease) or the end of study period, whichever occurred first.

Results

Participants were divided into six baseline BMI categories: < 18.5 (n = 55); 18.5–22.9 (n = 985); 23.0–24.9 (n = 1389); 25.0–29.9 (n = 3941); 30.0–34.9 (n = 1973); and ≥ 35.0 kg/m2 (n = 649). There were 38.9%, 23.5%, 24.7%, 8.4%, 2.7%, and 1.8% of patients experienced no-BW loss, initial BW loss of 0.0–2.4%, 2.5–4.9%, 5.0–7.4%, 7.5–9.9%, and ≥ 10.0%, associated with SGLT2i treatment, respectively. Compared with patients with normal BMI (BMI: 18.5–22.9 kg/m2), underweight (BMI: < 18.5 kg/m2) was associated with a higher risk of composite renal outcome (adjusted hazard ratio (aHR) [95% confidence intervals (CI)]: 2.17; [1.16–4.04]), whereas pre-obese (BMI: 25.0–29.9 kg/m2) associated with the lowest risk of composite renal outcome (0.52; [0.40–0.68]) after multivariate adjustment. Compared with those without BW loss after SGLT2i treatment, BW loss of 0.0–2.4% (0.55; [0.43–0.70]) and 2.5–4.9% (0.78; [0.63–0.98]) were associated with a lower risk, whereas BW loss ≥ 10.0% associated with a higher risk of composite renal outcome (1.61; [1.06–2.46]) after multivariate adjustment.

Conclusion

A modest BW loss of 0–5% associated with SGLT2i treatment was associated with a favorable renal outcome. Caution should be taken for whom are underweight at baseline or have a pronounced BW loss ≥ 10.0% associated with SGLT2i treatment, which was associated with a worse renal outcome.
Efficient conversion of phytosterols into 4-androstene-3,17-dione and its C1,2-dehydrogenized and 9α-hydroxylated derivatives by engineered Mycobacteria
Authors: Li Xin , Chen Tian , Peng Fei , Song Shikui , Yu Jingpeng , Sidoine Douanla Njimeli , Cheng Xiyao , Huang Yongqi , He Yijun , Su Zhengding
Published: August 2021

4-Androstene-3,17-dione (4-AD), 1,4-androstadiene-3,17-dione (ADD) and 9α-hydroxyl-4-androstene-3,17-dione (9OH-AD), which are important starting compounds for the synthesis of steroidal medicines, can be biosynthetically transformed from phytosterols by Mycobacterium strains. Genomic and metabolic analyses have revealed that currently available 4-AD-producing strains maintain the ability to convert 4-AD to ADD and 9OH-AD via 3-ketosteroid-1,2-dehydrogenase (KstD) and 3-ketosteroid-9α-hydroxylase (Ksh), not only lowering the production yield of 4-AD but also hampering its purification refinement. Additionally, these 4-AD industrial strains are excellent model strains to construct ADD- and 9OH-AD-producing strains. We recently found that Mycobacterium neoaurum HGMS2, a 4-AD-producing strain, harbored fewer kstd and ksh genes through whole-genomic and enzymatic analyses, compared with other strains (Wang et al. in Microbial Cell Fact 19:187, 2020). In this study, we attempted to construct an efficient 4-AD-producing strain by knocking out the kstd and ksh genes from the M. neoaurum HGMS2 strain. Next, we used kstd- and ksh-default HGMS2 mutants as templates to construct ADD- and 9OH-AD-producing strains by knocking in active kstd and ksh genes, respectively. We found that after knocking out its endogenous kstd and ksh genes, one of these knockout mutants, HGMS2Δkstd211 + ΔkshB122, showed a 20% increase in the rate of phytosterol to 4-AD conversion, compared relative to the wild-type strain and an increase in 4-AD yield to 38.3 g/L in pilot-scale fermentation. Furthermore, we obtained the ADD- and 9OH-AD-producing strains, HGMS2kstd2 + Δkstd211+ΔkshB122 and HGMS2kshA51 + Δkstd211+ΔkshA226, by knocking in heterogenous active kstd and ksh genes to selected HGMS2 mutants, respectively. During pilot-scale fermentation, the conversion rates of the ADD- and 9OH-AD-producing mutants transforming phytosterol were 42.5 and 40.3%, respectively, and their yields reached 34.2 and 37.3 g/L, respectively. Overall, our study provides efficient strains for the production of 4-AD, ADD and 9OH-AD for the pharmaceutical industry and provides insights into the metabolic engineering of the HGMS2 strain to produce other important steroidal compounds.
DDX52 knockdown inhibits the growth of prostate cancer cells by regulating c-Myc signaling
Authors: Yu Wandong , Ma Hangbin , Li Junhong , Ge Jinchao , Wang Pengyu , Zhou Yinghao , Zhang Jun , Shi Guowei
Published: August 2021

Background

DDX52 is a type of DEAD/H box RNA helicase that was identified as a novel prostate cancer (PCa) genetic locus and possible causal gene in a European large-scale transcriptome-wide association study. However, the functions of DDX52 in PCa remain undetermined. The c-Myc oncogene plays a crucial role in the development of PCa, but the factors that regulate the activity of c-Myc in PCa are still unknown.

Methods

We determined DDX52 protein levels in PCa tissues using immunohistochemistry (IHC). DDX52 expression and survival outcomes in other PCa cohorts were examined using bioinformatics analysis. The inhibition of DDX52 via RNA interference with shRNA was used to clarify the effects of DDX52 on PCa cell growth in vitro and in vivo. Gene set enrichment analysis and RNA sequencing were used to explore the signaling regulated by DDX52 in PCa. Western blotting and IHC were used to determine the possible DDX52 signaling mechanism in PCa.

Results

DDX52 expression was upregulated in PCa tissues. Bioinformatics analysis showed that the level of DDX52 further increased in advanced PCa, with a high DDX52 level indicating a poor outcome. In vitro and in vivo experiments showed that downregulating DDX52 impeded the growth of PCa cells. High DDX52 levels contributed to activating c-Myc signaling in PCa patients and PCa cells. Furthermore, DDX52 expression was regulated by c-Myc and positively correlated with c-Myc expression in PCa.

Conclusion

DDX52 was overexpressed in PCa tissues in contrast to normal prostate tissues. DDX52 knockdown repressed the growth of PCa cells in vitro and in vivo. Deleting c-Myc inhibited DDX52 expression, which affected the activation of c-Myc signaling.
Identification of a robust signature for clinical outcomes and immunotherapy response in gastric cancer: based on N6-methyladenosine related long noncoding RNAs
Authors: Han Tenghui , Xu Dong , Zhu Jun , Li Jipeng , Liu Lei , Deng Yanchun
Published: August 2021

Background

Gastric cancer (GC) is a globally prevalent cancer, ranking fifth for incidence and fourth for mortality worldwide. The N6-methyladenosine (m6A) related long noncoding RNAs (lncRNAs) were widely investigated in recent studies. Nevertheless, the underlying prognostic implication and tumor immune mechanism of m6A-related lncRNA in GC remain unknown.

Methods

We systematically assessed the m6A modification expression of 407 GC clinical samples based on 23 m6A regulators and comprehensively associated these genes with lncRNAs. Then, we constructed a m6A-related lncRNA prognostic signature (m6A-LPS) to evaluate both status and prognosis of the disease. Immune-related mechanisms were explored via dissecting tumor-infiltrating cells as well as applying tumor immune dysfunction and the exclusion algorithm. Furthermore, we validated the latent regulative mechanism of m6A-related lncRNA in GC cell lines.

Results

The m6A-LPS containing nine hub lncRNAs was built, which possessed a superior capability to predict the outcomes of GC patients. Meanwhile, we found an intimate correlation between the m6A-LPS and tumor infiltrating cells, and that the low-risk group had a higher expression of immune checkpoints and responsed more to immunotherapy than the high-risk group. Clinically, these crucial lncRNAs expression levels were verified in ten pairs of GC samples. In in vitro experiments, the abilities of migration and proliferation were significantly enhanced via downregulating the lncRNA AC026691.1. Both migrative and proliferative capabilities of tumor cells were significantly enhanced via downregulating the lncRNA AC026691.1. in vitro.

Conclusions

Collectively, the m6A-LPS could provide a novel prediction insight into the prognosis of GC patients and serve as an independent clinical factor for GC. These m6A-related lncRNAs might remodel the tumor microenvironment and affect the anti-cancer ability of immune checkpoint blockers. Importantly, lncRNA AC026691.1 could inhibit both migration and proliferation of GC by means of FTO regulation.
Bisphenol-A exposure and risk of breast and prostate cancer in the Spanish European Prospective Investigation into Cancer and Nutrition study
Authors: Salamanca-Fernández Elena , Rodríguez-Barranco Miguel , Amiano Pilar , Delfrade Josu , Chirlaque Maria Dolores , Colorado Sandra , Guevara Marcela , Jimenez Ana , Arrebola Juan Pedro , Vela Fernando , Olea Nicolás , Agudo Antonio , Sánchez Maria-José
Published: August 2021

Background

Bisphenol A (BPA) is an endocrine disruptor that it is present in numerous products of daily use. The aim of this study was to assess the potential association of serum BPA concentrations and the risk of incident breast and prostate cancer in a sub-cohort of the Spanish European Prospective Investigation into Cancer and Nutrition (EPIC).

Methods

We designed a case-cohort study within the EPIC-Spain cohort. Study population consisted on 4812 participants from 4 EPIC-Spain centers (547 breast cancer cases, 575 prostate cancer cases and 3690 sub-cohort participants). BPA exposure was assessed by means of chemical analyses of serum samples collected at recruitment. Borgan II weighted Cox regression was used to estimate hazard ratios.

Results

Median follow-up time in our study was 16.9 years. BPA geometric mean serum values of cases and sub-cohort were 1.12 ng/ml vs 1.10 ng/ml respectively for breast cancer and 1.33 ng/ml vs 1.29 ng/ml respectively for prostate cancer. When categorizing BPA into tertiles, a 40% increase in risk of prostate cancer for tertile 1 (p = 0.022), 37% increase for tertile 2 (p = 0.034) and 31% increase for tertile 3 (p = 0.072) was observed with respect to values bellow the limit of detection. No significant association was observed between BPA levels and breast cancer risk.

Conclusions

We found a similar percentage of detection of BPA among cases and sub-cohort from our population, and no association with breast cancer risk was observed. However, we found a higher risk of prostate cancer for the increase in serum BPA levels. Further investigation is needed to understand the influence of BPA in prostate cancer risk.
Park environment and moderate-to-vigorous physical activity in parks among adolescents in a high-density city: the moderating role of neighbourhood income
Authors: Zhang Ru , Zhang Chun-Qing , Lai Poh Chin , Cheng Wei , Schüz Benjamin , Kwan Mei-Po
Published: August 2021

Background

Urban parks are critical environmental resources in which adolescents engage in physical activity (PA). Evidence on the associations between park environmental characteristics and park-based PA in adolescents is mixed, particularly for high-density cities. Evidence is also lacking concerning the moderating role of neighbourhood socioeconomic status on the park-PA relationships. The current study aimed to examine the associations between park environmental characteristics and moderate-to-vigorous physical activity (MVPA) in parks among adolescents in Hong Kong and the moderating effect of neighbourhood income on these associations.

Methods

A cross-sectional study involving direct observations of adolescents was conducted in 32 randomly selected urban parks in Hong Kong. Park environmental characteristics were measured using the Community Park Audit Tool. Park-based MVPA among adolescents was measured using the System for Observation Play and Recreation in Communities. Neighbourhood income was extracted from the 2011 Hong Kong Population Census data on median household income.

Results

There was a significant positive association between the quality of amenities and park-based MVPA (metabolic equivalents per observation) in adolescents. However, the associations between the diversity of active facilities, greenness and adolescents’ park-based MVPA were not significant. Neighbourhood income moderated the association between adolescents’ park-based MVPA and park safety, where the relationship between park safety and park-based MVPA was significantly positive in low-income neighbourhoods but not significant in high-income neighbourhoods. An income-by-environment interaction was also observed concerning park aesthetics, with a negative relationship between park aesthetics and park-based MVPA in high-income neighbourhoods but not in low-income neighbourhoods.

Conclusion

Our findings provide evidence regarding how park environment and neighbourhood income impact adolescents’ park-based MVPA in Hong Kong. These findings can inform urban planning and policymakers who seek to improve urban park development in high-density cities.
Opioid overdose crises during the COVID-19 pandemic: implication of health disparities
Authors: Patel Ishika , Walter Lauren A. , Li Li
Published: August 2021

Background

Recent data suggest a disproportionate impact of opioid overdoses on Black Americans. The study aims to describe emergency department (ED) visits at a Southern, urban ED pertaining to opioid overdose and associated health disparities.

Methods

Patients presenting to the ED at the University of Alabama at Birmingham Hospital with opioid overdoses from January 1 to October 31, 2019, and from January 1 to October 31, 2020, were identified from electronic medical records.

Results

The total number of opioid overdose visits increased 9.7% (556 to 611) between January and October 2020 compared with 2019. Among patients who presented with opioid overdose, the mean ages were 50.3 years and 48.3 years, in 2019 and 2020, respectively. In both 2019 and 2020, more Blacks than whites were treated for opioid overdose in the ED (284 vs. 258 in 2019, and 306 vs. 271 in 2020) although 28 patients did not record their race in 2020. Consistently, more overdose deaths were observed in Blacks than in whites in 2020. More individuals seeking opioid overdose treatment were single in both years.

Conclusions

The study reported a greater number of visits for opioid overdoses from January to October of 2020 in an ED of a southeastern region, as well as higher overdose deaths in Blacks. Our findings highlight the importance of substance use treatment, harm reduction, and overdose prevention efforts that should be immediately present to reduce opioid overdose, especially for vulnerable populations in the South, i.e., Black community, and individuals experiencing singlehood.
Effects of LncRNA MEG3 on immunity and autophagy of non-small cell lung carcinoma through IDO signaling pathway
Authors: Wang Chuanqiang , Tao Xiangbo , Wei Jungong
Published: August 2021

Background

The study was done to investigate the effect of LncRNA MEG3 on the immunity and autophagy of non-small cell lung carcinoma through the IDO signaling pathway.

Methods

A total of 78 cases of early NSCLC patients (research group; RG) and 69 cases of health controls (control group; CG) during the same time were included. The contents of LncRNA MEG3 and miR-543 in peripheral blood and tissues and their diagnostic values for NSCLC were detected. The relationship between LncRNA MEG3 and miR-543 and their posttreatment contents and influence on the prognosis of NSCLC patients were tested. The expression of LncRNA MEG3, miR-543, and IDO (IDO1, IDO2, and TDO proteins) in the lung tissue of rats and the immune function in the CG and the RG were detected. The effects of LncRNA MEG3 and miR-543 on the biological behavior of NSCLC cells were determined. The role of LncRNA MEG3, miR-543, and IDO in NSCLC was verified.

Results

LncRNA MEG3 was low in peripheral blood and tissues, while miR-543 was high (P < 0.05); both had good diagnostic values for NSCLC (P < 0.05). LncRNA MEG3 had a negative correlation with miR-543 (P < 0.05) and influenced the prognosis of NSCLC patients (P < 0.05). LncRNA MEG3 in the lung tissue of rats using IDO inhibitor was elevated compared with that of lung carcinoma model rats (P < 0.05). The level of miR-543 was declined compared with that of lung carcinoma model rats (P < 0.05). The levels of IDO1, IDO2, and TDO proteins were evidently declined compared with those of lung carcinoma model rats (P < 0.05). Compared with lung carcinoma model rats, CD3+, CD4+, and CD4+/CD8+of IDO inhibitor rats were elevated, while CD8+was declined (P < 0.05). Cell proliferation and invasion ability and IDO1, IDO2, TDO, Beclin-1, and LC3-II proteins were declined in the sh-LncRNA MEG3 group (P < 0.05), while those in the mimics-miR-543 group were evidently elevated (P < 0.05). However, the double luciferase activity detection and RIP experiment confirmed that there was targeted regulation among them (P < 0.05).

Conclusion

MEG3 has low expression in NSCLC and affects the immunity and autophagy of NSCLC cells via regulating the miR-543/IDO signaling pathway, which is effective for the treatment of NSCLC.
Deep learning-based predictive biomarker of pathological complete response to neoadjuvant chemotherapy from histological images in breast cancer
Authors: Li Fengling , Yang Yongquan , Wei Yani , He Ping , Chen Jie , Zheng Zhongxi , Bu Hong
Published: August 2021

Background

Pathological complete response (pCR) is considered a surrogate endpoint for favorable survival in breast cancer patients treated with neoadjuvant chemotherapy (NAC). Predictive biomarkers of treatment response are crucial for guiding treatment decisions. With the hypothesis that histological information on tumor biopsy images could predict NAC response in breast cancer, we proposed a novel deep learning (DL)-based biomarker that predicts pCR from images of hematoxylin and eosin (H&E)-stained tissue and evaluated its predictive performance.

Methods

In total, 540 breast cancer patients receiving standard NAC were enrolled. Based on H&E-stained images, DL methods were employed to automatically identify tumor epithelium and predict pCR by scoring the identified tumor epithelium to produce a histopathological biomarker, the pCR-score. The predictive performance of the pCR-score was assessed and compared with that of conventional biomarkers including stromal tumor-infiltrating lymphocytes (sTILs) and subtype.

Results

The pCR-score derived from H&E staining achieved an area under the curve (AUC) of 0.847 in predicting pCR directly, and achieved accuracy, F1 score, and AUC of 0.853, 0.503, and 0.822 processed by the logistic regression method, respectively, higher than either sTILs or subtype; a prediction model of pCR constructed by integrating sTILs, subtype and pCR-score yielded a mean AUC of 0.890, outperforming the baseline sTIL-subtype model by 0.051 (0.839, P  =  0.001).

Conclusion

The DL-based pCR-score from histological images is predictive of pCR better than sTILs and subtype, and holds the great potentials for a more accurate stratification of patients for NAC.
Microglia are involved in phagocytosis and extracellular digestion during Zika virus encephalitis in young adult immunodeficient mice
Authors: Enlow William , Bordeleau Maude , Piret Jocelyne , Ibáñez Fernando González , Uyar Olus , Venable Marie-Christine , Goyette Nathalie , Carbonneau Julie , Tremblay Marie-Eve , Boivin Guy
Published: August 2021

Background

Zika virus (ZIKV) has been associated with several neurological complications in adult patients.

Methods

We used a mouse model deficient in TRIF and IPS-1 adaptor proteins, which are involved in type I interferon production, to study the role of microglia during brain infection by ZIKV. Young adult mice were infected intravenously with the contemporary ZIKV strain PRVABC59 (1 × 105PFUs/100 µL).

Results

Infected mice did not present overt clinical signs of the disease nor body weight loss compared with noninfected animals. However, mice exhibited a viremia and a brain viral load that were maximal (1.3 × 105genome copies/mL and 9.8 × 107genome copies/g of brain) on days 3 and 7 post-infection (p.i.), respectively. Immunohistochemistry analysis showed that ZIKV antigens were distributed in several regions of the brain, especially the dorsal hippocampus. The number of Iba1+/TMEM119+microglia remained similar in infectedversusnoninfected mice, but their cell body and arborization areas significantly increased in thestratum radiatumandstratum lacunosum-molecularelayers of the dorsal hippocampuscornu ammoni(CA)1, indicating a reactive state. Ultrastructural analyses also revealed that microglia displayed increased phagocytic activities and extracellular digestion of degraded elements during infection. Mice pharmacologically depleted in microglia with PLX5622 presented a higher brain viral load compared to untreated group (2.8 × 1010versus8.5 × 108genome copies/g of brain on day 10 p.i.) as well as an increased number of ZIKV antigens labeled with immunogold in the cytoplasm and endoplasmic reticulum of neurons and astrocytes indicating an enhanced viral replication. Furthermore, endosomes of astrocytes contained nanogold particles together with digested materials, suggesting a compensatory phagocytic activity upon microglial depletion.

Conclusions

These results indicate that microglia are involved in the control of ZIKV replication and/or its elimination in the brain. After depletion of microglia, the removal of ZIKV-infected cells by phagocytosis could be partly compensated by astrocytes.
Alpha-synuclein activates the classical complement pathway and mediates complement-dependent cell toxicity
Authors: Gregersen Emil , Betzer Cristine , Kim Woojin S. , Kovacs Gergo , Reimer Lasse , Halliday Glenda M. , Thiel Steffen , Jensen Poul Henning
Published: August 2021

Background

Synucleinopathies are characterized by neurodegeneration and deposition of the presynaptic protein α-synuclein in pathological protein inclusions. Growing evidence suggests the complement system not only has physiological functions in the central nervous system, but also is involved in mediating the pathological loss of synapses in Alzheimer’s disease. However, it is not established whether the complement system has a similar role in the diseases Parkinson's disease, Dementia with Lewy bodies, and multiple system atrophy (MSA) that are associated with α-synuclein aggregate pathology.

Methods

To investigate if the complement system has a pathological role in synucleinopathies, we assessed the effect of the complement system on the viability of an α-synuclein expressing cell model and examined direct activation of the complement system by α-synuclein in a plate-based activation assay. Finally, we investigated the levels of the initiator of the classical pathway, C1q, in postmortem brain samples from MSA patients.

Results

We demonstrate that α-synuclein activates the classical complement pathway and mediates complement-dependent toxicity in α-synuclein expressing SH-SY5Y cells. The α-synuclein-dependent cellular toxicity was rescued by the complement inhibitors RaCI (inhibiting C5) and Cp20 (inhibiting C3). Furthermore, we observed a trend for higher levels of C1q in the putamen of MSA subjects than that of controls.

Conclusion

α-Synuclein can activate the classical complement pathway, and the complement system is involved in α-synuclein-dependent cellular cytotoxicity suggesting the system could play a prodegenerative role in synucleinopathies.
Factors associated with testing for HIV and hepatitis C among behaviorally vulnerable men in Germany: a cross-sectional analysis upon enrollment into an observational cohort
Authors: Crowell Trevor A. , Qian Haoyu , Tiemann Carsten , Lehmann Clara , Boesecke Christoph , Stoehr Albrecht , Hartikainen Jukka , Esser Stefan , Bickel Markus , Spinner Christoph D. , Schneeweiß Stephan , Cordes Christiane , Brockmeyer Norbert , Jessen Heiko , Robb Merlin L. , Michael Nelson L. , Jansen Klaus , Streeck Hendrik ,   for the BRAHMS Study Team
Published: August 2021

Background

HIV and hepatitis C virus (HCV) have shared routes of transmission among men who have sex with men (MSM). Routine testing facilitates early diagnosis and treatment, thereby preventing morbidity and onward transmission. We evaluated factors associated with HIV and HCV testing in a behaviorally vulnerable cohort of predominantly MSM.

Methods

From June 2018 through June 2019, the BRAHMS study enrolled adults at ten German outpatient clinics that serve gender and sexual minority populations. Participants completed behavioral questionnaires that captured prior experience with HIV and HCV testing. Multivariable robust Poisson regression was used to evaluate factors potentially associated with testing in the previous 6 months.

Results

Among 1017 participants with median age 33 (interquartile range 28–39) years, 1001 (98.4%) reported any lifetime history of HIV testing and 787 (77.4%) reported any HCV testing, including 16 (1.6%) known to be living with HCV. Testing within the last 6 months was reported by 921 (90.6%) and 513 (50.4%) for HIV and HCV, respectively. Recent HIV testing was more common among participants with higher education level and recent HCV testing. Recent HCV testing was more common among participants with non-cisgender identity, lifetime history of illicit drug use, hepatitis B immunity or infection, and recent HIV testing.

Conclusion

Prior testing for HIV was common in this cohort, but interventions are needed to improve HCV risk stratification and access to testing. HIV testing infrastructure can be successfully leveraged to support HCV testing, but differentiated preventive care delivery is needed for some vulnerable populations.
Investigation of the clinical inter-observer bias in prostate fiducial marker image registration between CT and MR images
Authors: Persson Emilia , Emin Sevgi , Scherman Jonas , Jamtheim Gustafsson Christian , Brynolfsson Patrik , Ceberg Sofie , Gunnlaugsson Adalsteinn , Olsson Lars E.
Published: August 2021

Background and purpose

Inter-modality image registration between computed tomography (CT) and magnetic resonance (MR) images is associated with systematic uncertainties and the magnitude of these uncertainties is not well documented.
The purpose of this study was to investigate the potential uncertainty of gold fiducial marker (GFM) registration for localized prostate cancer and to estimate the inter-observer bias in a clinical setting.

Methods

Four experienced observers registered CT and MR images for 42 prostate cancer patients. Manual GFM identification was followed by a landmark-based registration. The absolute difference between observers in GFM identification and the displacement of the clinical target volume (CTV) was investigated. The CTV center of mass (CoM) vector displacements, DICE-index and Hausdorff distances for the observer registrations were compared against a clinical baseline registration. The time allocated for the manual registrations was compared.

Results

Absolute difference in GFM identification between observers ranged from 0.0 to 3.0 mm. The maximum CTV CoM displacement from the clinical baseline was 3.1 mm. Displacements larger than or equal to 1 mm, 2 mm and 3 mm were 46%, 18% and 4%, respectively. No statistically significant difference was detected between observers in terms of CTV displacement. Median DICE-index and Hausdorff distance for the CTV, with their respective ranges were 0.94 [0.70–1.00] and 2.5 mm [0.7–8.7].

Conclusions

Registration of CT and MR images using GFMs for localized prostate cancer patients was subject to inter-observer bias on an individual patient level. A CTV displacement as large as 3 mm occurred for individual patients. These results show that GFM registration in a clinical setting is associated with uncertainties, which motivates the removal of inter-modality registrations in the radiotherapy workflow and a transition to an MRI-only workflow for localized prostate cancer.
68Ga-DOTATATE PET-CT as a tool for radiation planning and evaluating treatment responses in the clinical management of meningiomas
Authors: Kowalski Emily S. , Khairnar Rahul , Gryaznov Anton A. , Kesari Vivek , Koroulakis Antony , Raghavan Prashant , Chen Wengen , Woodworth Graeme , Mishra Mark
Published: August 2021

Background and purpose

Meningiomas express the somatostatin receptor (SSTR), which normal bone and brain lack. PET imaging with SSTR ligands such as 68 Ga-DOTATATE have been recently shown to aid in the imaging and identification of menginiomas. We hypothesize that 68 Ga-DOTATATE PET/CT in conjunction with MRI aids in radiation (RT) target volume delineation and evaluating treatment response.

Materials and methods

Nineteen patients with meningiomas underwent 68 Ga-DOTATATE PET/CT and MRI for RT planning and/or post-treatment follow-up. Meningiomas were grade I (n = 9) or not biopsied (n = 8) and frequently involved base of skull (n = 10). Ten (53%) patients received post-operative RT and 9 (47%) received fractaionted RT. In the subgroup that underwent both pre- and post-RT 68 Ga-DOTATATE PET as well as MRI (n = 10), ROVER (ABX GmbH, Radeberg, Germany) adaptive thresholding software was utilized to measure total lesion activity (mean and max) before and after treatment. Tumor volume based on MRI was calculated before and after treatment. Total lesion activity and tumor volume changes were compared using Wilcoxon signed rank test.

Results

68 Ga-DOTATATE PET/CT identified intraosseous (n = 4, 22%), falcine (n = 5, 26%) and satellite lesions (n = 3, 19%) and clarified the diagnosis of meningioma, resulting in a change in management in three patients. Mean total lesion activity decreased 14.7% (median), from pre to post-RT 68 Ga-DOTATATE PET [range 97–8.5% (25–75%),S = − 26.5, p = 0.0039]. Max total lesion activity decreased 36% (median) over the same period [range 105–15% (25–75%), S = − 26.5 p = 0.0039]. In contrast, meningioma volumes based on MRI measurements did not significantly change per RECIST criteria and Wilcoxon signed rank test (S = − 3, p = 0.7422).

Conclusion

68 Ga-DOTATATE PET/CT helped confirm suspected diagnoses and delineate target volumes particularly when lesions involved osseous structures and the falx. Mean and max total tumor 68 Ga-DOTATATE activity on PET/CT decreased at three months following RT despite stable tumor volumes on MRI. Future studies are warranted to (1) assess the sensitivity and specificity of 68 Ga-DOTATATE PET/CT, (2) evaluate the impact of 68 Ga-DOTATATE PET/CT-based planning on treatment outcomes, and (3) assess the prognostic significance of these post-treatment imaging changes.
Projected soil organic carbon loss in response to climate warming and soil water content in a loess watershed
Authors: Zhao Fubo , Wu Yiping , Hui Jinyu , Sivakumar Bellie , Meng Xianyong , Liu Shuguang
Published: August 2021

Background

Soil organic carbon (SOC) plays a crucial role in the global carbon cycle and terrestrial ecosystem functions. It is widely known that climate change and soil water content (SWC) could influence the SOC dynamics; however, there are still debates about how climate change, especially climate warming, and SWC impact SOC. We investigated the spatiotemporal changes in SOC and its responses to climate warming and root-zone SWC change using the coupled hydro-biogeochemical model (SWAT-DayCent) and climate scenarios data derived under the three Representative Concentration Pathways (RCPs2.6, 4.5, and 8.5) from five downscaled Global Climate Models (GCMs) in a typical loess watershed––the Jinghe River Basin (JRB) on the Chinese Loess Plateau.

Results

The air temperature would increase significantly during the future period (2017–2099), while the annual precipitation would increase by 2.0–13.1% relative to the baseline period (1976–2016), indicating a warmer and wetter future in the JRB. Driven by the precipitation variation, the root-zone SWC would also increase (by up to 27.9% relative to the baseline under RCP4.5); however, the SOC was projected to decrease significantly under the future warming climate. The combined effects of climate warming and SWC change could more reasonably explain the SOC loss, and this formed hump-shaped response surfaces between SOC loss and warming-SWC interactions under both RCP2.6 and 8.5, which can help explain diverse warming effects on SOC with changing SWC.

Conclusions

The study showed a significant potential carbon source under the future warmer and wetter climate in the JRB, and the SOC loss was largely controlled by future climate warming and the root-zone SWC as well. The hump-shaped responses of the SOC loss to climate warming and SWC change demonstrated that the SWC could mediate the warming effects on SOC loss, but this mediation largely depended on the SWC changing magnitude (drier or wetter soil conditions). This mediation mechanism about the effect of SWC on SOC would be valuable for enhancing soil carbon sequestration in a warming climate on the Loess Plateau.
Development and validation of a novel blending machine learning model for hospital mortality prediction in ICU patients with Sepsis
Authors: Zeng Zhixuan , Yao Shuo , Zheng Jianfei , Gong Xun
Published: August 2021

Background

Early prediction of hospital mortality is crucial for ICU patients with sepsis. This study aimed to develop a novel blending machine learning (ML) model for hospital mortality prediction in ICU patients with sepsis.

Methods

Two ICU databases were employed: eICU Collaborative Research Database (eICU-CRD) and Medical Information Mart for Intensive Care III (MIMIC-III). All adult patients who fulfilled Sepsis-3 criteria were identified. Samples from eICU-CRD constituted training set and samples from MIMIC-III constituted test set. Stepwise logistic regression model was used for predictor selection. Blending ML model which integrated nine sorts of basic ML models was developed for hospital mortality prediction in ICU patients with sepsis. Model performance was evaluated by various measures related to discrimination or calibration.

Results

Twelve thousand five hundred fifty-eight patients from eICU-CRD were included as the training set, and 12,095 patients from MIMIC-III were included as the test set. Both the training set and the test set showed a hospital mortality of 17.9%. Maximum and minimum lactate, maximum and minimum albumin, minimum PaO2/FiO2 and age were important predictors identified by both random forest and extreme gradient boosting algorithm. Blending ML models based on corresponding set of predictors presented better discrimination than SAPS II (AUROC, 0.806 vs. 0.771; AUPRC 0.515 vs. 0.429) and SOFA (AUROC, 0.742 vs. 0.706; AUPRC 0.428 vs. 0.381) on the test set. In addition, calibration curves showed that blending ML models had better calibration than SAPS II.

Conclusions

The blending ML model is capable of integrating different sorts of basic ML models efficiently, and outperforms conventional severity scores in predicting hospital mortality among septic patients in ICU.
Recurrence of carcinoma showing thymus-like differentiation (CASTLE) involving the thyroid gland
Authors: Dang N. V. , Son L. X. , Hong N. T. T. , Nhung N. T. T. , Tung N. T. , Quang L. V.
Published: August 2021

Background

Carcinoma showing thymus-like differentiation (CASTLE) in the thyroid gland is a rare disease with generally a favorable prognosis. Treatment with surgery and adjuvant radiotherapy has been shown to improve local control and long-term survival rates. In this report, we present a case of a recurrent thyroid gland CASTLE and review the literature on the diagnosis and treatment of this disease.

Case presentation

A 60-year-old woman, who was diagnosed with a CASTLE thyroid tumor in 2015, had a total thyroidectomy and was maintained on thyroid hormone replacement (levothyroxine). After 5 years, the patient had a recurrence, in an advanced stage unsuitable for surgery. As the patient declined to undergo radiotherapy, she was followed up without intervention and is currently stable after 15 months.

Conclusions

CASTLE is a rare disease, diagnosed based on postoperative pathology and immunohistochemistry analysis, especially upon CD5 marker. In case of relapse, treatment options include surgery and radiotherapy; however conservative management without intervention is an acceptable alternative in some cases.