bluuebunny/crossref_metadata_2025 huggingface dataset:
A next-generation sequencing-based strategy combining microsatellite instability and tumor mutation burden for comprehensive molecular diagnosis of advanced colorectal cancer
Authors: Xiao Jian , Li Wenyun , Huang Yan , Huang Mengli , Li Shanshan , Zhai Xiaohui , Zhao Jing , Gao Chan , Xie Wenzhuan , Qin Hao , Cai Shangli , Bai Yuezong , Lan Ping , Zou Yifeng
Published: March 2021

Background

Mismatch repair (MMR)/microsatellite instability (MSI) and tumor mutational burden (TMB) are independent biomarkers that complement each other for predicting immune checkpoint inhibitors (ICIs) efficacy. Here we aim to establish a strategy that integrates MSI and TMB determination for colorectal cancer (CRC) in one single assay.

Methods

Surgical or biopsy specimens retrospectively collected from CRC patients were subjected to NGS analysis. Immunohistochemistry (IHC) and polymerase chain reaction (PCR) were also used to determine MMR/MSI for those having enough tissues. The NGS-MSI method was validated against IHC and PCR. The MSI-high (MSI-H) or microsatellite stable (MSS) groups were further stratified based on tumor mutational burden, followed by validation using the The Cancer Genome Atlas (TCGA) CRC dataset. Immune microenvironment was evaluated for each subgroup be profiling the expression of immune signatures.

Results

Tissues from 430 CRC patients were analyzed using a 381-gene NGS panel. Alterations in KRAS, NRAS, BRAF, and HER2 occurred at a significantly higher incidence among MSI-H tumors than in MSS patients (83.6% vs. 58.4%, p = 0.0003). A subset comprising 98 tumors were tested for MSI/MMR using all three techniques, where NGS proved to be 99.0 and 93.9% concordant with PCR and IHC, respectively. Four of the 7 IHC-PCR discordant cases had low TMB (1.1–8.1 muts/Mb) and were confirmed to have been misdiagnosed by IHC. Intriguingly, 4 of the 66 MSS tumors (as determined by NGS) were defined as TMB-high (TMB-H) using a cut-off of 29 mut/Mb. Likewise, 15 of the 456 MSS tumors in the TCGA CRC cohort were also TMB-H with a cut-off of 9 muts/Mb. Expression of immune signatures across subgroups (MSS-TMB-H, MSI-H-TMB-H, and MSS-TMB-L) confirmed that the microenvironment of the MSS-TMB-H tumors was similar to that of the MSI-H-TMB-H tumors, but significantly more immune-responsive than that of the MSS-TMB-L tumors, indicating that MSI combined with TMB may be more precise than MSI alone for immune microenvironment prediction.

Conclusion

This study demonstrated that NGS panel-based method is both robust and tissue-efficient for comprehensive molecular diagnosis of CRC. It also underscores the importance of combining MSI and TMB information for discerning patients with different microenvironment.
Artificial neural network model to predict post-hepatectomy early recurrence of hepatocellular carcinoma without macroscopic vascular invasion
Authors: Mai Rong-yun , Zeng Jie , Meng Wei-da , Lu Hua-ze , Liang Rong , Lin Yan , Wu Guo-bin , Li Le-qun , Ma Liang , Ye Jia-zhou , Bai Tao
Published: March 2021

Background

The accurate prediction of post-hepatectomy early recurrence (PHER) of hepatocellular carcinoma (HCC) is vital in determining postoperative adjuvant treatment and monitoring. This study aimed to develop and validate an artificial neural network (ANN) model to predict PHER in HCC patients without macroscopic vascular invasion.

Methods

Nine hundred and three patients who underwent curative liver resection for HCC participated in this study. They were randomly divided into derivation (n = 679) and validation (n = 224) cohorts. The ANN model was developed in the derivation cohort and subsequently verified in the validation cohort.

Results

PHER morbidity in the derivation and validation cohorts was 34.8 and 39.2%, respectively. A multivariable analysis revealed that hepatitis B virus deoxyribonucleic acid load, γ-glutamyl transpeptidase level, α-fetoprotein level, tumor size, tumor differentiation, microvascular invasion, satellite nodules, and blood loss were significantly associated with PHER. These factors were incorporated into an ANN model, which displayed greater discriminatory abilities than a Cox’s proportional hazards model, preexisting recurrence models, and commonly used staging systems for predicting PHER. The recurrence-free survival curves were significantly different between patients that had been stratified into two risk groups.

Conclusion

When compared to other models and staging systems, the ANN model has a significant advantage in predicting PHER for HCC patients without macroscopic vascular invasion.
Functional outcomes and complications of hook plate for bony mallet finger: a retrospective case series study
Authors: Wang Wei-Chih , Hsu Cheng-En , Yeh Chen-Wei , Ho Tsung-Yu , Chiu Yung-Cheng
Published: March 2021

Background

The treatment of mallet fracture using hook plate fixation was first introduced in 2007 and has subsequently shown excellent outcomes. Common complications, such as nail deformity and screw loosening, have also been reported. Very few studies have focused on these common complications or their prevention. In this study, we present the clinical outcomes and complications of our case series and describe the pitfalls and detailed solution of surgical tips to avoid common complications related to this procedure.

Methods

The retrospective case series of 16 patients with mallet fractures who underwent open reduction and hook plate fixation in our hospital from 2015 to 2020 were retrospectively reviewed. Data on extension lag, range-of-motion (ROM) of the distal interphalangeal joint (DIP) joint, the Disabilities of the Arm, Shoulder, and Hand (DASH) score, and surgical complications were collected and analysed. The clinical outcome was graded according to the Crawford mallet finger criteria.

Results

Sixteen patients were included in our analysis. The median DIP extension lag was 0° (range, 0° to 30°) and the median active DIP flexion angle was 60° (range, 40° to 90°). The median DASH score was 0 (range, 0–11.3). Fourteen patients with good and excellent results were satisfied with this treatment. The Complication rate in our patient series was 18%. Common complications reported in articles included wound necrosis, extension lag, nail deformity, and plate loosening.

Conclusions

Despite the fact that the treatment of mallet fracture with hook plate fixation has satisfactory functional outcomes, pitfalls, including iatrogenic nail germinal matrix injury, unnecessary soft tissue dissection, and insufficient screw purchase, were still reported. To avoid complications, we suggest modifications of the skin incision, soft tissue dissection, and screw position.
Analysis of IL-1β, CXCL8, and TNF-α levels in the crevicular fluid of patients with periodontitis or healthy implants
Authors: Aleksandrowicz Paweł , Brzezińska-Błaszczyk Ewa , Kozłowska Elżbieta , Żelechowska Paulina , Borgonovo Andrea Enrico , Agier Justyna
Published: March 2021

Background

Our study aimed to assess the level of IL-1β, CXCL8, and TNF-α in peri-implant sulcular fluid (PISF) collected from patients with no clinical symptoms of mucositis or peri-implantitis and compare them with cytokine concentration in gingival crevicular fluid (GCF) acquired from patients with healthy periodontium and those with varying severity of periodontitis.

Methods

A total of 189 subjects were included in the study, and GCF/PISF samples were checked for IL-1β, CXCL8, and TNF-α levels using an ELISA test.

Results

The IL-1β level in PISF in patients with implants was significantly lower than in GCF in patients with mild, moderate, or severe periodontitis. The CXCL8 level in PISF was considerably lower than in patients with moderate periodontitis. The TNF-α level in PISF in patients with implants was markedly higher compared to subjects with healthy periodontium or patients with mild periodontitis.

Conclusion

Analysis of cytokine levels may help describe the pathogenesis and early diagnosis of peri-implantitis and prevision in high-risk patients.
Use of complementary and alternative medicine in Norway: a cross-sectional survey with a modified Norwegian version of the international questionnaire to measure use of complementary and alternative medicine (I-CAM-QN)
Authors: Kristoffersen Agnete Egilsdatter , Quandt Sara A. , Stub Trine
Published: March 2021

Background

In recent decades complementary and alternative medicine (CAM) has been widely used worldwide as well as in Norway, where CAM is offered mainly outside the national health care service, mostly complementary to conventional treatment and fully paid for by the patients. With few exceptions, previous research has reported on frequency and associations of total CAM use in Norway rather than on single therapies and products. Therefore, in this present study we will map the use of CAM more precisely, including types of services, products, and self-help practices and further include reasons for use and helpfulness of the specific therapies used based on a modified Norwegian version of the I-CAM-Q (I-CAM-QN).

Method

Computer assisted telephone interviews using I-CAM-QN were conducted with 2001 randomly selected Norwegians aged 16 and above using multistage sampling in January 2019 with age and sex quotas for each area. Weights based on sex, age, education, and region corrected for selection biases, so that results are broadly representative of the Norwegian population. Descriptive statistics were carried out using Pearson’s Chi-square tests and t-tests to identify group differences.

Result

CAM use was reported by 62.2% of the participants during the prior12 months. Most participants had used natural remedies (47.4%), followed by self-help practices (29.1%) and therapies received from CAM providers (14.7%). Few of the participants had received CAM therapies from physicians (1.2%). Women were generally more likely to use CAM than men, younger people more likely than older, and participants with lower university education and income more likely than participants without university education, with higher university education and higher income. Mean number of visits per year to the different CAM providers ranged from 3.57 times to herbalists to 6.77 times to healers. Most of the participants found their use of CAM helpful.

Conclusion

This study confirms that CAM is used by a considerable segment of the Norwegian population. We suspect that the number of participants reporting CAM use is greater when specific therapies are listed in the questionnaire as a reminder (as in the I-CAM-QN) compared to more general questions about CAM use. The CAM modalities used are mainly received from CAM providers operating outside public health care or administered by the participants themselves.
Traditional and complementary medicine use among chronic haemodialysis patients: a nationwide cross-sectional study
Authors: Zakaria Nor Fadhlina , Mohd Noor Mohd Tawfeq , Abdullah Rafidah
Published: March 2021

Background

In the era of digital and improved conventional medicine, many continue to use traditional and complementary medicine (TCM). The prevalence of its usage is not well reported, especially in patients with end-stage kidney disease (ESKD) receiving haemodialysis, thus its benefits and adverse effects are not widely known. This study determines the prevalence, types, perceptions and factors associated with TCM use by chronic haemodialysis patients in Malaysia.

Methods

This is a multi-centre cross-sectional study involving patients undergoing haemodialysis treatment in Malaysia. A validated face-to-face questionnaire-based interview was conducted. Sociodemographic and clinical profiles of the patients, factors associated with TCM use, perceptions, sources of information, and disclosures to treating doctors were obtained. Data were analysed using SPSS software.

Results

A total of n = 329 participants were recruited. The mean age of the participants was 54.9 ± 12.5 years. The majority were Malays (72%) and females (54.7%). A total of 64.7% (n = 213) reported TCM use; n = 132 used TCM before the initiation of dialysis, while n = 81 used TCM after initiation. In the post-hoc analysis, patients who had never used TCM had a higher mean age (56.7 ± 12.3 years) than the patients who used TCM (51.1 ± 13.1) (p = 0.015) and were likely to have received primary education (p = 0.011). Unemployment was more likely to be associated with non-TCM use; with odds ratio 1.85 (95% CI: 1.15, 2.98). Biologically based therapy was found to be the most popular (97.2%) type of TCM, including herbal medicine (67.6%) and supplements (58.0%). Most respondents did not disclose their TCM use to their doctors (72.3%), and 41.8% had the perception that they felt better.

Conclusions

TCM is widely used among chronic haemodialysis patients in Malaysia, mainly herbal medicine and supplements. Non-disclosure to healthcare professionals and a poor monitoring and regulation of its use in ESKD patients could be detrimental. Awareness needs to be raised among healthcare professionals and the general population.

Trial registration

The Ethics Committee for Research, University Putra Malaysia (13th March 2019). Reference: UPM/TNCPI/RMC/1.4.18.2 (JKEUPM).
Variation of skeletal muscle ultrasound imaging intensity in horses after treadmill exercise: a proof of concept for glycogen content estimation
Authors: Tabozzi Sarah A. , Stancari Giovanni , Zucca Enrica , Tajoli Michela , Stucchi Luca , Lafortuna Claudio L. , Ferrucci Francesco
Published: March 2021

Background

Glycogen in skeletal muscle is a major source of energy during exercise and an important determinant of endurance capacity, so that its measurement may provide a meaningful marker of athletes’ preparation and a possible predictor of performance, both in humans and in equines. Gold standard of glycogen concentration measurement is the histochemical and biochemical analysis of biopsy-derived muscle tissue, an invasive and potentially injuring procedure. Recently, high-frequency ultrasound (US) technology is being exploited in human sports medicine to estimate muscle glycogen content. Therefore, aim of the present study is to evaluate the feasibility of US assessment of muscle glycogen in equines.

Results

US images ofgluteus medius(GL) andsemitendinosus(ST) muscles were obtained on eight healthy horses (3–10 years) before and after a steady-state exercise on treadmill (velocity: 4.0–12.5 m/s; duration: 2–20 min; heart rate: 137–218 b/min). Average image greyscale intensity was significantly different between GL and ST, both before and after exercise (p < 0.001). Comparing baseline and post-exercise US images, significant increase in greyscale intensity has been observed in ST (p < 0.001), but not in GL (p = 0.129). The volume of the exercise was significantly correlated with exercise-dependent change in image intensity (R2 = 0.891), consistent with a reduction of glycogen muscle stores resulting from aerobic activity.

Conclusions

US technique evidences also in horses muscle changes possibly associated to glycogen utilisation during exercise. Present results on a small sample need to be further confirmed and provide preliminary data warranting future validation by direct glycogen measurement through biopsy technique.
Nintedanib and immunomodulatory therapies in progressive fibrosing interstitial lung diseases
Authors: Cottin Vincent , Richeldi Luca , Rosas Ivan , Otaola Maria , Song Jin Woo , Tomassetti Sara , Wijsenbeek Marlies , Schmitz Manuela , Coeck Carl , Stowasser Susanne , Schlenker-Herceg Rozsa , Kolb Martin ,   the INBUILD Trial Investigators
Published: March 2021

Background

In the INBUILD trial in patients with chronic fibrosing interstitial lung diseases (ILDs) and a progressive phenotype, nintedanib reduced the rate of ILD progression with adverse events that were manageable for most patients. We investigated the potential impact of immunomodulatory therapies on the efficacy and safety of nintedanib.

Methods

Subjects with fibrosing ILDs other than idiopathic pulmonary fibrosis, who had shown progression of ILD within the prior 24 months despite management in clinical practice, were randomized to receive nintedanib or placebo. Certain immunomodulatory therapies were restricted for the first 6 months. We analyzed post-hoc the rate of decline in forced vital capacity (FVC) over 52 weeks in subgroups by glucocorticoid use at baseline and in analyses excluding subjects or FVC measurements taken after initiation of restricted immunomodulatory or antifibrotic therapies.

Results

Of 663 subjects, 361 (54.4%) were taking glucocorticoids at baseline (353 at a dose of ≤ 20 mg/day). In the placebo group, the adjusted rate of decline in FVC (mL/year) over 52 weeks was numerically greater in subjects taking than not taking glucocorticoids at baseline (− 206.4 [SE 20.2] vs − 165.8 [21.9]). The difference between the nintedanib and placebo groups was 133.3 (95% CI 76.6, 190.0) mL/year in subjects taking glucocorticoids at baseline and 76.1 (15.0, 137.2) mL/year in subjects who were not (interaction P = 0.18). The effect of nintedanib on reducing the rate of FVC decline in analyses excluding subjects or measurements taken after initiation of restricted immunomodulatory or antifibrotic therapies was similar to the primary analysis. The adverse event profile of nintedanib was similar between subjects who did and did not use prohibited or restricted therapies at baseline or during treatment with trial drug.

Conclusions

In patients with progressive fibrosing ILDs, the effect of nintedanib on reducing FVC decline was not influenced by the use of immunomodulatory therapies. Nintedanib can be used in combination with immunomodulatory therapies in patients with progressive fibrosing ILDs.

Trial registration ClinicalTrials.gov, NCT02999178. Registered 21 December 2016, https://clinicaltrials.gov/ct2/show/NCT02999178
Long non-coding RNA LINC01116 acts as an oncogene in prostate cancer cells through regulation of miR-744-5p/UBE2L3 axis
Authors: Yu Shengjie , Yu Huihong , Zhang Yuanfeng , Liu Chuan , Zhang Weili , Zhang Yunyun
Published: March 2021

Background

Long non-coding RNA (lncRNA) has been confirmed to exert a critical effect on the progression of tumors, including prostate cancer. Previous literature has demonstrated LINC01116 involves in activities of multiple cancers. However, the underlying role of LINC01116 in prostate cancer remains unclear.

Methods

qRT-PCR measured the expression of LINC01116 in prostate cancer cells. EdU experiment was used to detect cell proliferation. Transwell assays detected cell migration and invasion. Immunofluorescence staining and western blot assays were utilized to measure EMT progress. The binding relationship between RNAs was confirmed by a series of mechanism assays. In addition, rescue experiments were conducted to verify the relationship among RNAs.

Results

LINC01116 was found to be highly expressed in prostate cancer cells. Functional assays indicated that inhibition of LINC01116 could suppress cell proliferation, migration, invasion and EMT progress. Also, miR-744-5p was proven to bind with LINC01116. Moreover, UBE2L3 was verified as the target gene of miR-744-5p. In rescue assays, we discovered that inhibited miR-744-5p or overexpressed UBE2L3 could offset the suppressive influence of silencing LINC01116 on prostate cancer cells.

Conclusion

Our study suggested that lncRNA LINC01116 acted as an oncogene in prostate cancer and accelerated prostate cancer cell growth through regulating miR-744-5p/UBE2L3 axis.
Deregulation of angiopoietin-like 4 slows ovarian cancer progression through vascular endothelial growth factor receptor 2 phosphorylation
Authors: Wu Yuxian , Gao Jinghai , Liu Xiaojun
Published: March 2021

Background

As a tissue-specific proangiogenic or antiangiogenic agent, angiopoietin-like 4 (ANGPTL4) has recently gained attention in many diseases, such as metabolic syndrome, cardiovascular disease and cancer. However, the roles of ANGPTL4 in angiogenesis and tumor growth in epithelial ovarian cancer, the most lethal gynecologic malignancy, remain unclear.

Objective

To identify a novel mechanism of ANGPTL4 inhibition in epithelial ovarian cancer.

Methods

Western blot, quantitative reverse transcription PCR, and immunofluorescence analyses were applied to evaluate ANGPTL4 expression in ovarian cancer cell lines. Cell proliferation, migration, and invasion were investigated through 5-ethynyl-2′-deoxyuridine (EdU) incorporation, CCK-8 and Transwell assays. The expression of epithelial-mesenchymal transition (EMT)-related proteins in ovarian cancer cells and tumor-bearing mice was evaluated. CD31 staining was used to identify tumor angiogenesis. Immunoprecipitation was performed to examine the regulatory relationship between ANGPTL4 and the vascular endothelial growth factor receptor 2 (VEGFR2)/vascular endothelial (VE)-cadherin/Src complex. VEGFR2 phosphorylation at Y949 and VE-cadherin expression were assessed by western blotting. Inactivation of VEGFR2 Y949 phosphorylation was achieved in a MISIIR-TAg VEGFR2Y949F/Y949F mouse model.

Results

Here, we demonstrated that ANGPTL4 was overexpressed in A2780 and CAOV3 ovarian cancer cells. In vitro assays indicated that inhibition of ANGPTL4 by lentiviral small interfering RNA does not alter ovarian cancer cell proliferation, migration, invasion, and EMT, while ANGPTL4 silencing exhibited significant inhibitory effects on tumor angiogenesis, growth, and metastasis in vivo. Immunoprecipitation analysis showed that suppression of ANGPTL4 was accompanied by dissociation of the VEGFR2/VE-cadherin/Src complex and phosphorylation of VEGFR2 Y949 in A2780 and CAOV3 ovarian tumors. Inactivation of VEGFR2 Y949 phosphorylation in MISIIR-TAg VEGFR2Y949F/Y949F mice abolished all tumor-suppressive effects of ANGPTL4 inhibition in spontaneous ovarian carcinoma.

Conclusions

Overall, our results indicate that ANGPLT4 silencing delays tumor progression in specific types of ovarian cancer and may be a potential target for individualized treatment of ovarian cancer.
Energy and protein intake in medical and geriatric inpatients with MEDPass versus conventional administration of oral nutritional supplements: study protocol for the randomized controlled MEDPass Trial
Authors: Kurmann Silvia , Reber Emilie , Vasiloglou Maria F. , Schuetz Philipp , Schoenenberger Andreas W. , Uhlmann Katja , Sterchi Anna-Barbara , Stanga Zeno
Published: March 2021

Background

Disease-related malnutrition is highly prevalent in hospitalized medical and geriatric inpatients. It is associated with negative outcomes such as muscle wasting, decline of functional status, and increased morbidity and mortality. Oral nutritional supplements (ONS) are frequently used in nutritional therapy to increase intake. However, compliance to ONS is often limited and maybe improved by prescribing ONS in small portions timed with the medication (MEDPass). However, it is unknown whether the MEDPass administration enhances patients’ total energy and protein intake.

Methods

The MEDPass Trial is a randomized, controlled, open-label superiority trial. Patients in the MEDPass group receive 50 ml of ONS four times per day, distributed with the medication rounds. Patients in the control group receive ONS between meals. The primary outcome is average daily energy intake (% of calculated daily requirement). For our power analysis, we assumed that administration of ONS in the MEDPass administration mode increases energy intake by at least 10% (i.e., by 200 kcal for an average energy requirement of 2200 kcal/day). Thus, with the inclusion of 200 patients, this trial has 80% power to demonstrate that intervention group patients have an average intake of 2200 kcal/day (SD 500 kcal) versus 2000 kcal/day (SD 500 kcal) in control group patients. Energy and protein intakes from ONS and all food consumed are monitored continuously throughout the hospital stay and are statistically compared to the patient’s requirements. Secondary outcomes include average daily protein intake (% of calculated daily requirement), average intake of ONS/day, the course of body weight, handgrip strength, appetite, and nausea. Furthermore, hospital length of stay and 30-day mortality are assessed. The primary statistical analysis will be performed as an intention-to-treat analysis adjusted for the stratification factors used in randomization.

Discussion

To our knowledge, this is the first randomized controlled trial assessing total energy and protein intake for the entire hospitalization period in patients receiving MEDPass versus conventional ONS administration. Thus, the MEDPass Trial will fill a gap and answer this relevant clinical question.

Trial registration

ClinicalTrials.gov NCT03761680. Registered on 3 December 2018. Kofam.ch SNCTP000003191. Registered on 15 October 2018
Effectiveness of low-dose theophylline for the management of biomass-associated COPD (LODOT-BCOPD): study protocol for a randomized controlled trial
Authors: Siddharthan Trishul , Pollard Suzanne L. , Jackson Peter , Robertson Nicole M. , Wosu Adaeze C. , Rahman Nihaal , Padalkar Roma , Sekitoleko Isaac , Namazzi Esther , Alupo Patricia , Hurst John R. , Kalyesubula Robert , Dowdy David , Wise Robert , Barnes Peter J. , Checkley William , Kirenga Bruce
Published: March 2021

Background

COPD is a leading cause of death globally, with the majority of morbidity and mortality occurring in low- and middle-income country (LMIC) settings. While tobacco-smoke exposure is the most important risk factor for COPD in high-income settings, household air pollution from biomass smoke combustion is a leading risk factor for COPD in LMICs. Despite the high burden of biomass smoke-related COPD, few studies have evaluated the efficacy of pharmacotherapy in this context. Currently recommended inhaler-based therapy for COPD is neither available nor affordable in most resource-limited settings. Low-dose theophylline is an oral, once-a-day therapy, long used in high-income countries (HICs), which has been proposed for the management of COPD in LMICs in the absence of inhaled steroids and/or bronchodilators. The Low-dose Theophylline for the Management of Biomass-Associated COPD (LODOT-BCOPD) trial investigates the clinical efficacy and cost-effectiveness of low-dose theophylline for the management of biomass-related COPD in a low-income setting.

Methods

LODOT-BCOPD is a randomized, double-blind, placebo-controlled trial to test the efficacy of low-dose theophylline in improving respiratory symptoms in 110 participants with moderate to severe COPD in Central Uganda. The inclusion criteria are as follows: (1) age 40 to 80 years, (2) full-time resident of the study area, (3) daily biomass exposure, (4) post-bronchodilator FEV1/FVC below the 5th percentile of the Global Lung Initiative mixed ethnic reference population, and (5) GOLD Grade B-D COPD. Participants will be randomly assigned to receive once daily low-dose theophylline (200 mg ER, Unicontin-E) or placebo for 52 weeks. All participants will receive education about self-management of COPD and rescue salbutamol inhalers. We will measure health status using the St. George’s Respiratory Questionnaire (SGRQ) and quality of life using the EuroQol-5D (EQ-5D) at baseline and every 6 months. In addition, we will assess household air pollution levels, serum inflammatory biomarkers (fibrinogen, hs-CRP), and theophylline levels at baseline, 1 month, and 6 months. The primary outcome is change in SGRQ score at 12 months. Lastly, we will assess the cost-effectiveness of the intervention by calculating quality-adjusted life years (QALYs) from the EQ-5D.

Trial registration

ClinicalTrials.gov NCT03984188. Registered on June 12, 2019

Trial acronym

Low-dose Theophylline for the Management of Biomass-Associated COPD (LODOT-BCOPD)
Reproductive injury in male BALB/c mice infected with Neospora caninum
Authors: Li Hang , Yang Bing-Yi , Liu Ming-Ming , Zhao Shao-Wei , Xie Su-Zhu , Wang Hao , Zhang Shuang , Xuan Xue-Nan , Jia Li-Jun
Published: March 2021

Background

Neospora caninum is one of the main causes of abortion in pregnant animals. However, N. caninum-induced reproductive injury in male mice is still unclear.

Methods

Male BALB/c mice were infected with a bovine isolate of N. caninum, and the organ coefficients of the testis and epididymis were measured. Lesions in the testis and epididymis were observed by light microscopy and transmission electron microscopy. Expression of the spermatogenic cell apoptosis-related proteins p53 and caspase-3 was detected by western blot. The expression of spermatogenesis-related genes in the testis was detected by reverse transcription-PCR. Sperm morphology and motility were observed. The levels of nitric oxide (NO) and antisperm antibody (AsAb) in the testicular homogenates and hormones in the serum were detected by enzyme-linked immunosorbent assay. The reproductive capacity of the male mice was detected using a reproduction test.

Results

The organ coefficients of the testis and epididymis of the experimental group were significantly downregulated. Light microscopy examination revealed that the spermatogenic cells of the testis were arranged in a disordered manner, and the number was reduced. The number of sperm in the epididymal lumen was significantly reduced, and the cytoplasm exhibited vacuolation and degeneration. Ultrastructural studies revealed that the cells of the testis and epididymis tissues showed varying degrees of disease. The level of p53 and caspase-3 expression in the testis was significantly upregulated. The expression of the testicular spermatogenesis-related genes Herc4, Ipo11 and Mrto4 were strongly downregulated. Observation of sperm by microscopic examination revealed significantly reduced sperm density and sperm motility, and the number of sperm deformities was significantly increased. The level of NO and AsAb was significantly increased. The levels of luteinizing hormone, follicle-stimulating hormone and gonadotropin-releasing hormone were significantly upregulated, whereas the levels of testosterone, thyrotropin-releasing hormone, thyroxine and thyroid-stimulating hormone were significantly downregulated. After challenge, the infected male mice and healthy female mice were caged together: the subsequent fetal death rate was increased, and the conception rate, litter size, number of live births and the birth weight were significantly reduced.

Conclusions

Infection of male BALB/c mice with the bovine isolate of N. caninum induced varying degrees of injury to the testis, epididymis and sperm of the mice, destroyed spermatogenesis and affected the reproductive capacity.

Graphical Abstract
EGFL6 promotes colorectal cancer cell growth and mobility and the anti‐cancer property of anti-EGFL6 antibody
Authors: Sung Ting-Yi , Huang Han-Li , Cheng Chun-Chun , Chang Fu-Ling , Wei Po-Li , Cheng Ya-Wen , Huang Cheng-Chiao , Lee Yu-Ching , HuangFu Wei-Chun , Pan Shiow-Lin
Published: March 2021

Background

The availability of a reliable tumor target for advanced colorectal cancer (CRC) therapeutic approaches is critical since current treatments are limited. Epidermal growth factor-like domain 6 (EGFL6) has been reported to be associated with cancer development. Here, we focused on the role of EGFL6 in CRC progression and its clinical relevance. In addition, an anti-EGFL6 antibody was generated by phage display technology to investigate its potential therapeutic efficacy in CRC.

Results

EGFL6 expression
significantly increased in the colon tissues from CRC patients and mice showing
spontaneous tumorigenesis, but not in normal tissue. Under hypoxic condition, EGFL6
expression was enhanced at both protein and transcript levels. Moreover, EGFL6
could promote cancer cell migration invasion, and proliferation of CRC cells via up-regulation of
the ERK/ AKT pathway. EGFL6 also regulated cell migration, invasion,
proliferation, and self-renewal through EGFR/αvβ3 integrin receptors. Treatment
with the anti-EGFL6 antibody EGFL6-E5-IgG showed tumor-inhibition and
anti-metastasis abilities in the xenograft and syngeneic mouse models,
respectively. Moreover, EGFL6-E5-IgG treatment had no adverse effect on
angiogenesis and wound healing

Conclusions

We demonstrated that EGFL6 plays a role in CRC tumorigenesis and tumor progression, indicating that EGFL6 is a potential therapeutic target worth further investigation.
When the genome bluffs: a tandem duplication event during generation of a novel Agmo knockout mouse model fools routine genotyping
Authors: Sailer Sabrina , Coassin Stefan , Lackner Katharina , Fischer Caroline , McNeill Eileen , Streiter Gertraud , Kremser Christian , Maglione Manuel , Green Catherine M. , Moralli Daniela , Moschen Alexander R. , Keller Markus A. , Golderer Georg , Werner-Felmayer Gabriele , Tegeder Irmgard , Channon Keith M. , Davies Benjamin , Werner Ernst R. , Watschinger Katrin
Published: March 2021

Background

Genome editing in mice using either classical approaches like homologous recombination or CRISPR/Cas9 has been reported to harbor off target effects (insertion/deletion, frame shifts or gene segment duplications) that lead to mutations not only in close proximity to the target site but also outside. Only the genomes of few engineered mouse strains have been sequenced. Since the role of the ether-lipid cleaving enzyme alkylglycerol monooxygenase (AGMO) in physiology and pathophysiology remains enigmatic, we created a knockout mouse model for AGMO using EUCOMM stem cells but unforeseen genotyping issues that did not agree with Mendelian distribution and enzyme activity data prompted an in-depth genomic validation of the mouse model.

Results

We report a gene segment tandem duplication event that occurred during the generation of an Agmo knockout-first allele by homologous recombination. Only low homology was seen between the breakpoints. While a single copy of the recombinant 18 kb cassette was integrated correctly around exon 2 of the Agmo gene, whole genome nanopore sequencing revealed a 94 kb duplication in the Agmo locus that contains Agmo wild-type exons 1–3. The duplication fooled genotyping by routine PCR, but could be resolved using qPCR-based genotyping, targeted locus amplification sequencing and nanopore sequencing. Despite this event, this Agmo knockout mouse model lacks AGMO enzyme activity and can therefore be used to study its physiological role.

Conclusions

A duplication event occurred at the exact locus of the homologous recombination and was not detected by conventional quality control filters such as FISH or long-range PCR over the recombination sites. Nanopore sequencing provides a cost convenient method to detect such underrated off-target effects, suggesting its use for additional quality assessment of gene editing in mice and also other model organisms.
Direct and indirect evidence of efficacy and safety of rapid exercise tests for exertional desaturation in Covid-19: a rapid systematic review
Authors: Kalin Asli , Javid Babak , Knight Matthew , Inada-Kim Matt , Greenhalgh Trisha
Published: March 2021

Background

Even when resting pulse oximetry is normal in the patient with acute Covid-19, hypoxia can manifest on exertion. We summarise the literature on the performance of different rapid tests for exertional desaturation and draw on this evidence base to provide guidance in the context of acute Covid-19.

Main research questions

What exercise tests have been used to assess exertional hypoxia at home or in an ambulatory setting in the context of Covid-19 and to what extent have they been validated?

What exercise tests have been used to assess exertional hypoxia in other lung conditions, to what extent have they been validated and what is the applicability of these studies to acute Covid-19?

Method

AMED, CINAHL, EMBASE MEDLINE, Cochrane and PubMed using LitCovid, Scholar and Google databases were searched to September 2020. Studies where participants had Covid-19 or another lung disease and underwent any form of exercise test which was compared to a reference standard were eligible. Risk of bias was assessed using QUADAS 2. A protocol for the review was published on the Medrxiv database.

Results

Of 47 relevant papers, 15 were empirical studies, of which 11 described an attempt to validate one or more exercise desaturation tests in lung diseases other than Covid-19. In all but one of these, methodological quality was poor or impossible to fully assess. None had been designed as a formal validation study (most used simple tests of correlation). Only one validation study (comparing a 1-min sit-to-stand test [1MSTST] with reference to the 6-min walk test [6MWT] in 107 patients with interstitial lung disease) contained sufficient raw data for us to calculate the sensitivity (88%), specificity (81%) and positive and negative predictive value (79% and 89% respectively) of the 1MSTST. The other 4 empirical studies included two predictive studies on patients with Covid-19, and two on HIV-positive patients with suspected pneumocystis pneumonia. We found no studies on the 40-step walk test (a less demanding test that is widely used in clinical practice to assess Covid-19 patients). Heterogeneity of study design precluded meta-analysis.

Discussion

Exertional desaturation tests have not yet been validated in patients with (or suspected of having) Covid-19. A stronger evidence base exists for the diagnostic accuracy of the 1MSTST in chronic long-term pulmonary disease; the relative intensity of this test may raise safety concerns in remote consultations or unstable patients. The less strenuous 40-step walk test should be urgently evaluated.
Murrayanine suppresses the proliferation and metastasis of human breast cancer cells via induction of apoptosis and inhibition of RANK/RANKL signaling pathway
Authors: zhang Jingnan , Wang Feng , Qu Gengbao , Wang Baokai
Published: March 2021

Murrayanine and its derivatives have been shown to exhibit anticancer activities against different types of human cancer cells. However, the effects of murrayanine on the proliferation and metastasis of breast cancer cells are yet to be studied. The present study was designed to evaluate the effects of murrayanine on the proliferation and metastasis of human breast cancer via regulation of RANK/RANKL pathway. The results showed RANK/RANKL pathway to be highly activated in human breast cancer tissues and cell lines. However, treatment of the CAMA-1 breast cancer cells with murrayanine (0, 9, 18 and 36 μM) caused a significant (P < 0.05) decline in the expression of RANK, RANKL and OPG in CAMA-1 cells. Additionally, murrayanine inhibited the growth of CAMA-1 cells with an IC50 of 18 μM. The antiproliferative of murrayanine were found be due to its ability to inhibit the expression of RANK, RANKL and OPG in CAMA-1 cells. To unveil if murrayanine exerted its effects via inhibition of RANK/RANKL pathway, the expression of RNAK was knocked down in CAMA-1 cells. It was found that murrayanine and RANK silencing both inhibited the growth CAMA-1 cells via induction of apoptosis. Additionally, murrayanine and RANK silencing both inhibited the migration, invasion and epithelial to mesenchymal transition of the CAMA-1 cells. Taken together, murrayanine exhibits significant anticancer activity against the breast cancer cells via induction of apoptosis and inhibition of RANK/RANKL signaling pathway. These findings suggest that murrayanine may prove to be a beneficial lead molecule for the development of breast cancer chemotherapy.
Barley somatic embryogenesis-an attempt to modify variation induced in tissue culture
Authors: Orłowska Renata
Published: March 2021

Background

Somatic embryogenesis is a phenomenon carried out in an environment that generates abiotic stress. Thus, regenerants may differ from the source of explants at the morphological, genetic, and epigenetic levels. The DNA changes may be the outcome of induction media ingredients (i.e., copper and silver ions) and their concentrations and time of in vitro cultures.

Results

This study optimised the level of copper and silver ion concentration in culture media parallel with the induction medium longevity step towards obtaining barley regenerants via somatic embryogenesis with a minimum or maximum level of tissue culture-induced differences between the donor plant and its regenerants. The optimisation process is based on tissue culture-induced variation evaluated via the metAFLP approach for regenerants derived under varying in vitro tissue culture conditions and exploited by the Taguchi method. In the optimisation and verification experiments, various copper and silver ion concentrations and the different number of days differentiated the tested trials concerning the tissue culture-induced variation level, DNA demethylation, and de novo methylation, including symmetric (CG, CHG) and asymmetric (CHH) DNA sequence contexts. Verification of optimised conditions towards obtaining regenerants with minimum and maximum variability compared to donor plants proved useful. The main changes that discriminate optimised conditions belonged to DNA demethylation events with particular stress on CHG context.

Conclusions

The combination of tissue culture-induced variation evaluated for eight experimental trials and implementation of the Taguchi method allowed the optimisation of the in vitro tissue culture conditions towards the minimum and maximum differences between a source of tissue explants (donor plant) and its regenerants from somatic embryos. The tissue culture-induced variation characteristic is mostly affected by demethylation with preferences towards CHG sequence context.
Optimising medication management in children and young people with ADHD using a computerised test (QbTest): a feasibility randomised controlled trial
Authors: Williams Laura , Hall Charlotte L. , Brown Susan , Guo Boliang , James Marilyn , Franceschini Matilde , Clarke Julie , Selby Kim , Vijayan Hena , Kulkarni Neeta , Brown Nikki , Sayal Kapil , Hollis Chris , Groom Madeleine J.
Published: March 2021

Background

Medication for attention deficit hyperactivity disorder (ADHD) should be closely monitored to ensure optimisation. There is growing interest in using computerised assessments of ADHD symptoms to support medication monitoring. The aim of this study was to assess the feasibility and acceptability of a randomised controlled trial (RCT) to evaluate the efficacy of one such computerised assessment, the Quantified Behavior (Qb) Test, as part of medication management for ADHD.

Methods

This feasibility multi-site RCT conducted in child and adolescent mental health and community paediatric settings recruited participants aged 6–15 years diagnosed with ADHD starting stimulant medication. Participants were randomised into one of two arms: experimental (QbTest protocol) where participants completed a QbTest at baseline and two follow-up QbTests on medication (2–4 weeks and 8–10 weeks later) and control where participants received treatment as usual, including at least two follow-up consultations. Measures of parent, teacher, and clinician-rated symptoms and global functioning were completed at each time point. Clinicians recorded treatment decision-making and health economic measures were obtained. Data were analysed using multi-level modelling and participants (children and parents) and clinicians were interviewed about their experiences, resulting data were thematically analysed.

Results

Forty-four children and young people were randomised. Completion of study outcome measures by care-givers and teachers ranged from 52 to 78% at baseline to 47–65% at follow-up. Participants reported the questionnaires to be useful to complete. SNAP-IV inattention scores showed greater reduction in the intervention than the control group (− 5.85, 95% CI − 10.33, − 1.36,). Engagement with the intervention ranged from 100% at baseline, to 78% follow-up 1 and 57% follow-up 2. However, only 37% of QbTests were conducted in the correct time period. Interview data highlighted that the objectivity of the QbTest was appreciated by clinicians and parents. Clinicians commented that the additional time and resources required meant that it is not feasible to use QbTest for all cases.

Conclusion

The trial design and protocol appear to be feasible and acceptable but could be improved by modifying QbTest time periods and the method of data collection. With these changes, the protocol may be appropriate for a full trial. Adding QbTest may improve symptom outcome as measured by SNAP-IV.

Trial registration

ClinicalTrials.gov, NCT03368573, prospectively registered, 11th December 2017, and ISRCTN, ISRCTN69461593, retrospectively registered, 10th April 2018
Preretinal hemorrhage preventing subretinal involvement in multilayer macular hemorrhages: coincidence or plausible relationship?
Authors: Nascimento Priscila A. , Carbajo Cristina N. , Mendonça Luisa S. M. , Okimoto Juliana T. , Barros Bogdan C. M. , Abucham-Neto Julio Z.
Published: March 2021

Background

To report four cases with interesting anatomical presentations of multilayer macular hemorrhages with preretinal hemorrhage possibly preventing subretinal involvement of the macular area.

Cases presentation

Observational study of four patients presenting with macular hemorrhages.

Results

Four patients with multilayer macular hemorrhage due to different causes, presented with a halo-shaped submacular hemorrhage coincident with the preretinal hemorrhage borders. After resolution, in all cases, the macular area underneath the preretinal hemorrhage was found to be spared.

Conclusion

We hypothesized that an extensive preretinal hemorrhage can exert a mechanical force pushing the subretinal hemorrhage towards the periphery, consequently protecting the macular area.
Pathogenicity of two fungal pathogens on larvae of the elm leaf beetle, Xanthogaleruca luteola Muller (Coleoptera: Chrysomelidae)
Authors: Ebrahimifar Jafar , Jamshidnia Arsalan
Published: March 2021

Background

The elm leaf beetle (ELB), Xanthogaleruca luteola Muller (Coleoptera: Chrysomelidae), is one of the most important insect pests that cause damage and physiological stress for the trees. In the present research, pathogenicity of both entomopathogenic fungi (EPF), Beauveria bassiana and Metarhizium anisopliae on the 1st, 2nd, and 3rd instar larval of ELB at the concentrations of 105, 106, 107, and 108 conidia per ml, using immersion and spray methods and were evaluated.

Results

The results showed that LC50 of B. bassiana in immersion and spray methods were 3.88 × 102 and 7.52 × 102 conidia/ml, while LC50 of M. anisopliae in mentioned methods were 7.05 × 102 and 11.85 × 102 conidia/ml, respectively. The highest mortality rate in both EPF was observed in the 1st instar larvae that were significantly different than the control treatment. Moreover, the highest and lowest mortality in both EPF, using immersion and spray methods, were observed at the concentrations of 108 and 105 conidia/ml, respectively. Therefore, susceptibility of 1st and 2nd larval instars was more than 1st instar larvae. Based on the findings of the present study, there was no mortality during the first 24 h, while after 2 days, the mortality was recorded and increased over time in case of both pathogens. Accordingly, B. bassiana and M. anisopliae were pathogenic on larvae of ELB.

Conclusions

The outputs of the study demonstrated that applications of these eco-friendly fungal pathogens with other non-chemical methods are recommended. Of course, under natural conditions, their efficiency may change; thus, more comprehensive studies are necessary.