bluuebunny/crossref_metadata_2025 huggingface dataset:
CRISPR-targetedMAGT1insertion restores XMEN patient hematopoietic stem cells and lymphocytes
Authors: Brault Julie , Liu Taylor , Bello Ezekiel , Liu Siyuan , Sweeney Colin L. , Meis Ronald J. , Koontz Sherry , Corsino Cristina , Choi Uimook , Vayssiere Guillaume , Bosticardo Marita , Dowdell Kennichi , Lazzarotto Cicera R. , Clark Aaron B. , Notarangelo Luigi D. , Ravell Juan C. , Lenardo Michael J. , Kleinstiver Benjamin P. , Tsai Shengdar Q. , Wu Xiaolin , Dahl Gary A. , Malech Harry L. , De Ravin Suk See
Published: June 2021

XMEN disease, defined as “X-linked MAGT1 deficiency with increased susceptibility to Epstein-Barr virus infection and N-linked glycosylation defect,” is a recently described primary immunodeficiency marked by defective T cells and natural killer (NK) cells. Unfortunately, a potentially curative hematopoietic stem cell transplantation is associated with high mortality rates. We sought to develop an ex vivo targeted gene therapy approach for patients with XMEN using a CRISPR/Cas9 adeno-associated vector (AAV) to insert a therapeutic MAGT1 gene at the constitutive locus under the regulation of the endogenous promoter. Clinical translation of CRISPR/Cas9 AAV-targeted gene editing (GE) is hampered by low engraftable gene-edited hematopoietic stem and progenitor cells (HSPCs). Here, we optimized GE conditions by transient enhancement of homology-directed repair while suppressing AAV-associated DNA damage response to achieve highly efficient (>60%) genetic correction in engrafting XMEN HSPCs in transplanted mice. Restored MAGT1 glycosylation function in human NK and CD8+ T cells restored NK group 2 member D (NKG2D) expression and function in XMEN lymphocytes for potential treatment of infections, and it corrected HSPCs for long-term gene therapy, thus offering 2 efficient therapeutic options for XMEN poised for clinical translation.
Uncovering the roles of microRNAs/lncRNAs in characterising breast cancer subtypes and prognosis
Authors: Li Xiaomei , Truong Buu , Xu Taosheng , Liu Lin , Li Jiuyong , Le Thuc D.
Published: June 2021

Background

Accurate prognosis and identification of cancer subtypes at molecular level are important steps towards effective and personalised treatments of breast cancer. To this end, many computational methods have been developed to use gene (mRNA) expression data for breast cancer subtyping and prognosis. Meanwhile, microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) have been extensively studied in the last 2 decades and their associations with breast cancer subtypes and prognosis have been evidenced. However, it is not clear whether using miRNA and/or lncRNA expression data helps improve the performance of gene expression based subtyping and prognosis methods, and this raises challenges as to how and when to use these data and methods in practice.

Results

In this paper, we conduct a comparative study of 35 methods, including 12 breast cancer subtyping methods and 23 breast cancer prognosis methods, on a collection of 19 independent breast cancer datasets. We aim to uncover the roles of miRNAs and lncRNAs in breast cancer subtyping and prognosis from the systematic comparison. In addition, we created an R package, CancerSubtypesPrognosis, including all the 35 methods to facilitate the reproducibility of the methods and streamline the evaluation.

Conclusions

The experimental results show that integrating miRNA expression data helps improve the performance of the mRNA-based cancer subtyping methods. However, miRNA signatures are not as good as mRNA signatures for breast cancer prognosis. In general, lncRNA expression data does not help improve the mRNA-based methods in both cancer subtyping and cancer prognosis. These results suggest that the prognostic roles of miRNA/lncRNA signatures in the improvement of breast cancer prognosis needs to be further verified.
Study on the differences of gene expression between pear and apple wild cultivation materials based on RNA-seq technique
Authors: Zhang Huangwei , Li Meng , Kong Min , Dunwell Jim M. , Zhang Yuyan , Yue Chao , Wu Juyou , Zhang Shaoling
Published: June 2021

Background

Pears and apples are both perennial deciduous trees of theRosaceaefamily, and both are important economic fruit trees worldwide. The emergence of many varieties in the market has been mostly domesticated from wild to cultivated and regulated by the differential expression of genes. However, the molecular process and pathways underlying this phenomenon remain unclear. Four typical wild and cultivar pear and apple trees at three developmental stages were used in our study to investigate the molecular process at the transcriptome level.

Result

Physiological observations indicated the obvious differences of size, weight, sugar acid content and peel color in wild and cultivar fruit among each developmental stage. Using next-generation sequencing based RNA-seq expression profiling technology, we produced a transcriptome in procession of a large fraction of annotated pear and apple genes, and provided a molecular basis underlying the phenomenon of wild and cultivar fruit tree differences. 5921 and 5744 differential expression genes were identified in pear and apple at three developmental stages respectively. We performed temporal and spatial differential gene expression profiling in developing fruits. Several key pathways such as signal transduction, photosynthesis, translation and many metabolisms were identified as involved in the differentiation of wild and cultivar fruits.

Conclusion

In this study, we reported on the next-generation sequencing study of the temporal and spatial mRNA expression profiling of pear and apple fruit trees. Also, we demonstrated that the integrated analysis of pear and apple transcriptome, which strongly revealed the consistent process of domestication inRosaceaefruit trees. The results will be great influence to the improvement of cultivar species and the utilization of wild resources.
Lack of association between vitamin D insufficiency and clinical outcomes of patients with COVID-19 infection
Authors: Davoudi Alireza , Najafi Narges , Aarabi Mohsen , Tayebi Atefeh , Nikaeen Roja , Izadyar Hamideh , Salar Zahra , Delavarian Leila , Vaseghi Narges , Daftarian Zahra , Ahangarkani Fatemeh
Published: June 2021

Background

A protective effect of vitamin D against COVID-19 infection is under investigation. We aimed to analyze the effect of vitamin D sufficiency on the clinical outcomes of patients infected with COVID-19.

Methods

In this cross-sectional study we analyzed the vitamin D levels of COVID-19 patients who were admitted to Razi Hospital (an infectious disease referral center in Mazandaran province in northern Iran) from February to March 2020. Overall, a cutoff point of 30 ng/mL was used for the definition of vitamin D sufficiency.

Results

One hundred fifty-three patients were analyzed in this study who had laboratory documentation of a 25(OH) D level at the time of hospitalization. The vitamin D levels of the patients were 27.19 ± 20.17 ng/mL. In total, 62.7% (n = 96) of the patients had a 25(OH) D level of less than 30 ng/mL and 37.25% (n = 57) had a 25(OH) D level of more than 30 ng/mL. In total, 49% (n = 75) of the patients suffered from at least one underlying disease. The univariate and multivariable regression showed that vitamin D sufficiency was not associated with a statistically significant lower risk of adverse clinical outcomes of COVID-19 such as duration of hospitalization and severity of infection (P > 0.05).

Conclusions

Sufficient vitamin D levels were not found to be protective against adverse clinical outcomes in patients infected with COVID-19. Chronic disorders in COVID-19 patients were found to have greater relevance than vitamin D levels in determining the adverse outcomes of the infection. Further studies are needed to determine the role of vitamin D level in predicting the outcomes of COVID-19 infection.
Therapeutic method for early-stage second primary non-small lung cancer: analysis of a population-based database
Authors: Chen Congcong , Wu Zixiang , Wu Ziheng , Wu Chuanqiang , Wang Qi , Zhan Tianwei , Dong Lingjun , Fang Shuai , Wu Ming
Published: June 2021

Background

Early-stage non-small lung cancer patients may survive long enough to develop second primary lung cancers. However, few studies have accurately described the therapeutic method, evaluation or prognostic factors for long-term survival in this complex clinical scenario.

Methods

Patients who had first and second primary non-small lung cancer in the Surveillance, Epidemiology, and End Results database between 2004 and 2015 were evaluated. Patients were included when their tumors were pathologically diagnosed as non-small lung cancer and in the early-stage (less than 3 cm and with no lymph node metastasis). Therapeutic methods were categorized as lobectomy, sublobectomy or no surgery. The influence of different therapeutic methods on the overall survival rate was compared.

Results

For the first primary tumor, patients who underwent lobectomy achieved superior survival benefits compared with patients who underwent sublobectomy. For the second primary tumor, long-term survival was similar in patients who underwent lobectomy and those who underwent sublobectomy treatment. The multivariate analysis indicated that age, disease-free time interval, sex, and first and second types of surgery were independent prognostic factors for long-term survival. Our results showed that the 5-year overall survival rate was 91.9% when the disease-free interval exceeded 24 months.

Conclusion

Lobectomy for the first primary tumor followed by sublobectomy for the second primary tumor may be a beneficial therapeutic method for patients. If the disease-free interval exceeds 24 months, the second primary tumor will have no influence on the natural course for patients diagnosed with a first primary non-small lung cancer.
To measure the amount of ocular deviation in strabismus patients with an eye-tracking virtual reality headset
Authors: Yeh Po-Han , Liu Chun-Hsiu , Sun Ming-Hui , Chi Sheng-Chu , Hwang Yih-Shiou
Published: June 2021

Purpose

To investigate the accuracy of a newly developed, eye-tracking virtual reality (VR)-based ocular deviation measurement system in strabismus patients.

Methods

A VR-based ocular deviation measurement system was designed to simulate the alternative prism cover test (APCT). A fixation target was made to alternate between two screens, one in front of each eye, to simulate the steps of a normal prism cover test. Patient’s eye movements were recorded by built-in eye tracking. The angle of ocular deviation was compared between the APCT and the VR-based system.

Results

This study included 38 patients with strabismus. The angle of ocular deviation measured by the VR-based system and the APCT showed good to excellent correlation (intraclass correlation coefficient, ICC = 0.897 (range: 0.810–0.945)). The 95% limits of agreement was 11.32 PD. Subgroup analysis revealed a significant difference between esotropia and exotropia (p < 0.001). In the esotropia group, the amount of ocular deviation measured by the VR-based system was greater than that measured by the APCT (mean = 4.65 PD), while in the exotropia group, the amount of ocular deviation measured by the VR-based system was less than that of the APCT (mean = − 3.01 PD). The ICC was 0.962 (range: 0.902–0.986) in the esotropia group and 0.862 (range: 0.651–0.950) in the exotropia group. The 95% limits of agreement were 6.62 PD and 11.25 PD in the esotropia and exotropia groups, respectively.

Conclusions

This study reports the first application of a consumer-grade and commercial-grade VR-based device for assessing angle of ocular deviation in strabismus patients. This device could provide measurements with near excellent correlation with the APCT. The system also provides the first step to digitize the strabismus examination, as well as the possibility for its application in telemedicine.
Coughing children in family practice and primary care: a systematic review of prevalence, aetiology and prognosis
Authors: Bergmann Milena , Haasenritter Jörg , Beidatsch Dominik , Schwarm Sonja , Hörner Kaja , Bösner Stefan , Grevenrath Paula , Schmidt Laura , Viniol Annika , Donner-Banzhoff Norbert , Becker Annette
Published: June 2021

Background

For evidence-based decision making, primary care physicians need to have specific and reliable information on the pre-test probabilities of underlying diseases and a symptom’s course. We performed a systematic review of symptom-evaluating studies in primary care, following three research questions: (1) What is the prevalence of the symptom cough in children consulting primary care physicians? (2) What are the underlying aetiologies of cough and the respective frequencies? (3) What is the prognosis of children with cough?

Methods

Following a pre-defined algorithm and independent double reviewer ratings we searched MEDLINE and EMBASE. All quantitative original research articles in English, French or German were included if they focused on unselected study populations of children consulting a primary care physician for cough. We used the random effects model for meta-analysis in subgroups, if justifiable in terms of heterogeneity.

Results

We identified 14 eligible studies on prevalence, five on aetiology and one on prognosis. Prevalence estimates varied between 4.7 and 23.3% of all reasons for an encounter, or up to estimates of 60% when related to patients or consultations. Cough in children is more frequent than in adults, with lowest prevalences in adolescents and in summer. Acute cough is mostly caused by upper respiratory tract infections (62.4%) and bronchitis (33.3%); subacute or chronic cough by recurrent respiratory tract infection (27.7%), asthma (up to 50.4% in cough persisting more than 3 weeks), and pertussis (37.2%). Potentially serious diseases like croup, pneumonia or tuberculosis are scarce. In children with subacute and chronic cough the total duration of cough ranged from 24 to 192 days. About 62.3% of children suffering from prolonged cough are still coughing two months after the beginning of symptoms.

Conclusion

Cough is one of the most frequent reasons for an encounter in primary care. Our findings fit in with current guideline recommendations supporting a thoughtful wait-and-see approach in acute cough and a special awareness in chronic cough of the possibility of asthma and pertussis. Further evidence of aetiological pre-test probabilities is needed to assess the diagnostic gain based on patient history and clinical signs for differential diagnoses of cough in children.
The role of sickness absence diagnosis for the risk of future inpatient- or specialized outpatient care in a Swedish population-based twin sample
Authors: Ropponen Annina , Wang Mo , Narusyte Jurgita , Kärkkäinen Sanna , Blom Victoria , Svedberg Pia
Published: June 2021

Background

Studies of consequences of sickness absence such as health and well-being have been rare whereas risk factors for sickness absence have been studied extensively. This study assumed the consequences of sickness absence would differ by diagnostic group or by patient care type. The aim was to investigate sickness absence due to various diagnosis groups as a predictor for subsequent inpatient- and specialized outpatient care while controlling for familial confounding.

Methods

We utilized the register data of 69,552 twin individuals between 16 and 80 years of age (48% women). The first incident sickness absence spell, from baseline year 2005, including diagnosis of sickness absence was our primary exposure of interest and we followed them until the first incident inpatient- and specialized outpatient care episode with main diagnosis code or until 31.12.2013.

Results

A total of 7464 incident sickness absence spells took place (11%), 42% had inpatient care and 83% specialized outpatient care (mean follow-up time 3.2 years, SD 3.1 years). All the main sickness absence diagnosis groups were associated with increased risk of future care in comparison to no sickness absence. Controlling for confounders attenuated the associations in magnitude but with retaining direction, and we could not confirm an effect of familial factors.

Conclusions

Sickness absence predicts both inpatient- and specialized outpatient care and the association is universal across diagnosis groups. The lower survival time and incidence rates of inpatient than specialized outpatient care point towards severity of diseases assumption. This finding was also universal across sickness absence diagnosis groups.
Do weight perception and bullying victimization account for links between weight status and mental health among adolescents?
Authors: Patte Karen A. , Livermore Maram , Qian Wei , Leatherdale Scott T.
Published: June 2021

Background

The purpose of this study was to explore whether the way youth perceive their weight and their experiences of bullying victimization account for the increased risk of depression and anxiety symptoms, and poor psychosocial well-being, associated with overweight/obesity in a large sample of Canadian secondary school students. We also explored if associations differed by gender.

Methods

We used cross-sectional survey data from year 7 (2018–19) of the COMPASS study. The sample included 57,059 students in grades 9–12 (Secondary III-V in Quebec) at 134 Canadian secondary schools (Alberta, British Columbia, Ontario, Quebec). First, multiple regression models tested associations between body mass index (BMI) classification and mental health outcomes (anxiety [GAD-7] and depression [CESD-10] symptoms, and psychosocial well-being [Diener’s Flourishing Scale]). Second, weight perception and bullying victimization were added to the models. Models were stratified by gender and controlled for sociodemographic covariates and school clustering.

Results

When weight perception and bullying victimization were added to the models, obesity BMI status no longer predicted internalizing symptoms and flourishing scores relative to normal-weight BMIs. Students with ‘overweight’ or ‘underweight’ perceptions, and experiences of bullying victimization in the past month, reported higher anxiety and depressive symptomatology, and lower flourishing levels, in comparison to students with ‘about right’ weight perceptions and without experiences of bullying victimization, respectively, controlling for BMI status. Results were largely consistent across boys and girls.

Conclusions

Results suggest perceptions of weight and experiences of bullying independently contribute to differences in mental health outcomes by weight status among youth. Continued efforts targeting weight-based bullying and weight bias, and the promotion of body size acceptance and positive body image, may help reduce the risk of mental illness and poor mental health among adolescents.
Risk and protective factors for anxiety during COVID-19 pandemic
Authors: Zhong Jingyi , Zhong Chenghui , Qiu Lan , Li Jiayi , Lai Jiayi , Lu Wenfeng , Wang Shuguang , Zhong Jiacai , Zhao Jing , Zhou Yun
Published: June 2021

Background

Coronavirus Disease 2019 (COVID-19) is a global pandemic and an anxiety-provoking event. There are few studies to identify potential risk and protective factors related to anxiety during COVID-19 pandemic.

Methods

We collected information on demographic data and lifestyles by a web-based survey of 19,802 participants from 34 provinces in China during COVID-19 pandemic. Level of anxiety was evaluated using the Self-Rating Anxiety Scale. We used ordinal multivariable logistic regression to estimate the associations of anxiety level with potential risk and protective factors. We further developed a new score to simplify the assessment of anxiety during COVID-19 crisis.

Results

Among 19,802 participants, we found that those who were front-line medical personnel, suffered from chronic disease, with present symptoms of SARS-CoV-2 infection or contact history had 112, 93, 40 and 15% increased risk of higher anxiety level; while those with knowledge about personal protective measures or wore masks had 75 and 29% lower risk of higher anxiety level respectively. We developed a risk score by calculating the sum of single score of 17 factors. Each one increase of the risk score was associated with a 297% increase in anxiety index score. In categorical analysis, low risk (the risk score between 1 to 2), the moderate risk group (the risk score of 3) and high risk group (the risk score ≥ 4) had − 0.40 (95% CI: − 1.55, 0.76), 1.44 (95% CI: 0.27, 2.61) and 9.18 (95% CI: 8.04, 10.33) increase in anxiety index score, and 26% (95% CI: − 7, 72%), 172% (95% CI: 100, 270%), and 733% (95% CI: 516, 1026%) higher risk of anxiety respectively, when compared with the very low risk group (the risk score of 0). The AUC was 0.73 (95% CI, 0.72, 0.74) for the model fitted the developed risk score, with the cut-off point of 3.5.

Conclusions

These findings revealed protective and risk factors associated with anxiety, and developed a simple method of identifying people who are at an increased risk of anxiety during COVID-19 pandemic.
Suspension sutures facilitate single-incision laparoscopic-assisted rectal pull-through for Hirschsprung disease
Authors: Nguyen Liem Thanh , Nguyen Anh Tho , Nguyen Quang Thanh , Tran Quynh Anh , Bui Hau Duc , Pham Hien Duy
Published: June 2021

Background

To present a surgical technique of single-incision laparoscopic-assisted endorectal pull-through (SILEP) with suspension sutures using conventional instruments for Hirschsprung disease (HD) and its long-term follow-up outcomes.

Methods

The procedure began with a 1 cm transumbilical skin incision. Three separate punctures were made in the fascia with a 5 mm scope in the middle and 5 mm and 3 mm ports for working instruments on the left and right, respectively. The first suspension suture was placed to secure the sigmoid colon to the abdominal wall. A window was created through the rectal mesentery, and dissection around the rectum was carried out. The second suspension suture was performed to suspend the rectovesical peritoneal fold or the rectovaginal peritoneal fold to the abdominal wall. Dissection around the rectum was continued downward to approximately 1 cm below the peritoneal fold. Then, the operation was completed by a transanal approach.

Results

Forty patients underwent SILEP from March 2013 to April 2015. The median age was 2.7 months (ranging from 1 to 17 months). The mean operative time was 96 ± 23 min. No conversion to an open operation was required. The average hospitalization time was 4.5 ± 2 days. There were no intraoperative or perioperative complications. Long-term follow-up results were obtained from 38 patients. A frequency of defecation from every other day to twice a day was noted for 33 patients (86.8%) and more often for 5 patients (13.2%). Two patients had enterocolitis (5.2%).

Conclusion

Single-incision laparoscopic rectal pull-through with suspension sutures using conventional instruments is feasible and safe for HD with good long-term outcomes.
Low bone mineral density following gastric bypass is not explained by lifestyle and lack of exercise
Authors: Stevens Katharina , Hultin Hella , Hellman Per , Sundbom Magnus
Published: June 2021

Background

Bariatric surgery, Roux-en-Y gastric bypass (RYGBP) in particular, is associated with weight loss as well as low bone mineral density. Bone mineral density relies upon multiple factors, some of which are lifestyle factors. The aim of this study was to compare lifestyle factors in order to eliminate them as culprits of the suspected difference in BMD in RYGBP operated and controls.

Materials and methods

Study participants included 71 RYGBP-operated women (42.3 years, BMI 33.1 kg/m2) and 94 controls (32.4 years, BMI 23.9 kg/m2). Each completed a DEXA scan, as well as survey of lifestyle factors (e.g. physical activity in daily life, corticosteroid use, and calcium intake). All study participants were premenopausal Caucasian women living in the same area. Blood samples were taken in RYGBP-patients.

Results

BMD was significantly lower in RYGBP, femoral neck 0.98 vs. 1.04 g/cm2 compared to controls, despite higher BMI (present and at 20 years of age) and similar physical activity and calcium intake. In a multivariate analysis, increased time since surgery and age were negatively associated with BMD of the femoral neck and total hip in RYGBP patients.

Conclusion

Despite similar lifestyle, RYGBP was followed by a lower BMD compared to controls. Thus, the reduced BMD in RYGBP cannot be explained, seemingly nor prevented, by lifestyle factors. As the reduction in BMD was associated with time since surgery, strict follow-up is a lifelong necessity after bariatric surgery, and especially important in younger bariatric patients.
The effect of Humanitude care methodology on improving empathy: a six-year longitudinal study of medical students in Japan
Authors: Fukuyasu Yusuke , Kataoka Hitomi U. , Honda Miwako , Iwase Toshihide , Ogawa Hiroko , Sato Masaru , Watanabe Mayu , Fujii Chikako , Wada Jun , DeSantis Jennifer , Hojat Mohammadreza , Gonnella Joseph S.
Published: June 2021

Background

Empathy, which involves understanding another person’s experiences and concerns, is an important component for developing physicians’ overall competence. This longitudinal study was designed to test the hypothesis that medical students’ empathy can be enhanced and sustained by Humanitude Care Methodology, which focuses on perception, emotion and speech.

Methods

This six-year longitudinal observational study examined 115 students who entered Okayama University Medical School in 2013. The study participants were exposed to two empathy-enhancing programs: (1) a communication skills training program (involving medical interviews) and (2) a Humanitude training program aimed at enhancing their empathy. They completed the Jefferson Scale of Empathy (JSE) seven times: when they entered medical school, before participation in the first program (medical interview), immediately after the first program, before the second program (Humanitude exercise), immediately after the second program, and in the 5th and 6th year (last year) of medical school. A total of 79 students (69% of the cohort) completed all seven test administrations of the JSE.

Results

The mean JSE scores improved significantly after participation in the medical interview program (p < 0.01) and the Humanitude training program (p = 0.001). However, neither program showed a sustained effect.

Conclusions

The Humanitude training program as well as medical interview training program, had significant short-term positive effects for improving empathy among medical students. Additional reinforcements may be necessary for a long-term sustained effect.
Membrane recruitment of Atg8 by Hfl1 facilitates turnover of vacuolar membrane proteins in yeast cells approaching stationary phase
Authors: He Cheng-Wen , Cui Xue-Fei , Ma Shao-Jie , Xu Qin , Ran Yan-Peng , Chen Wei-Zhi , Mu Jun-Xi , Li Hui , Zhu Jing , Gong Qingqiu , Xie Zhiping
Published: June 2021

Background

The vacuole/lysosome is the final destination of autophagic pathways, but can also itself be degraded in whole or in part by selective macroautophagic or microautophagic processes. Diverse molecular mechanisms are involved in these processes, the characterization of which has lagged behind those of ATG-dependent macroautophagy and ESCRT-dependent endosomal multivesicular body pathways.

Results

Here we show that as yeast cells gradually exhaust available nutrients and approach stationary phase, multiple vacuolar integral membrane proteins with unrelated functions are degraded in the vacuolar lumen. This degradation depends on the ESCRT machinery, but does not strictly require ubiquitination of cargos or trafficking of cargos out of the vacuole. It is also temporally and mechanistically distinct from NPC-dependent microlipophagy. The turnover is facilitated by Atg8, an exception among autophagy proteins, and an Atg8-interacting vacuolar membrane protein, Hfl1. Lack of Atg8 or Hfl1 led to the accumulation of enlarged lumenal membrane structures in the vacuole. We further show that a key function of Hfl1 is the membrane recruitment of Atg8. In the presence of Hfl1, lipidation of Atg8 is not required for efficient cargo turnover. The need for Hfl1 can be partially bypassed by blocking Atg8 delipidation.

Conclusions

Our data reveal a vacuolar membrane protein degradation process with a unique dependence on vacuole-associated Atg8 downstream of ESCRTs, and we identify a specific role of Hfl1, a protein conserved from yeast to plants and animals, in membrane targeting of Atg8.
Mechanochemical preparation of chrysomycin A self-micelle solid dispersion with improved solubility and enhanced oral bioavailability
Authors: Xu Zhuomin , Zheng Shanshan , Gao Xin , Hong Yulu , Cai Yue , Zhang Qiuqin , Xiang Jiani , Xie Dehui , Song Fuxing , Zhang Huawei , Wang Hong , Sun Xuanrong
Published: June 2021

Background

Chrysomycin A (CA) has been reported as numerous excellent biological activities, such as antineoplastic and antibacterial. Though, poor solubility of CA limited its application in medical field. Due to good amphiphilicity and potential anticancer effect of disodium glycyrrhizin (Na2GA) as an excipient, an amorphous solid dispersion (Na2GA/CA-BM) consisting of CA and Na2GA was prepared in the present study by mechanochemical technology (roll mill ML-007, zirconium balls, 30 rpm, 2.5 h) to improve the solubility and oral bioavailability of CA. Then, Na2GA/CA-BM was self-assembled to micelles in water. The interaction of CA and Na2GA in solid state were investigated by X-ray diffraction studies, polarized light microscopy, and scanning electron microscope. Meanwhile, the properties of the sample solution were analyzed by dynamic light scattering and transmission electron. Furthermore, the oral bioavailability and antitumor ability of Na2GA/CA-BM in vivo were tested, providing a theoretical basis for future application of CA on cancer therapy.

Results

CA encapsulated by Na2GA was self-assembled to nano-micelles in water. The average diameter of nano-micelle was 131.6 nm, and zeta potential was − 11.7 mV. Three physicochemical detections showed that CA was transformed from crystal into amorphous form after treated with ball milling and the solubility increased by 50 times. Na2GA/CA-BM showed a significant increase of the bioavailability about two time that of free CA. Compared with free CA, the in-vivo antitumor studies also exhibited that Na2GA/CA-BM had an excellent inhibition of tumor growth.

Conclusions

Na2GA/CA-BM nanoparticles (131.6 nm, − 11.7 mV) prepared by simple and low-cost mechanochemical technology can improve oral bioavailability and antitumor efficacy of CA in vivo, suggesting a potential formulation for efficient anticancer treatment.

Graphic abstract
The phosphatase Shp1 interacts with and dephosphorylates cortactin to inhibit invadopodia function
Authors: Varone Alessia , Amoruso Chiara , Monti Marcello , Patheja Manpreet , Greco Adelaide , Auletta Luigi , Zannetti Antonella , Corda Daniela
Published: June 2021

Background

Invadopodia are actin-based cell-membrane protrusions associated with the extracellular matrix degradation accompanying cancer invasion. The elucidation of the molecular mechanisms leading to invadopodia formation and activity is central for the prevention of tumor spreading and growth. Protein tyrosine kinases such as Src are known to regulate invadopodia assembly, little is however known on the role of protein tyrosine phosphatases in this process. Among these enzymes, we have selected the tyrosine phosphatase Shp1 to investigate its potential role in invadopodia assembly, due to its involvement in cancer development.

Methods

Co-immunoprecipitation and immunofluorescence studies were employed to identify novel substrate/s of Shp1AQ controlling invadopodia activity. The phosphorylation level of cortactin, the Shp1 substrate identified in this study, was assessed by immunoprecipitation, in vitro phosphatase and western blot assays. Short interference RNA and a catalytically-dead mutant of Shp1 expressed in A375MM melanoma cells were used to evaluate the role of the specific Shp1-mediated dephosphorylation of cortactin. The anti-invasive proprieties of glycerophosphoinositol, that directly binds and regulates Shp1, were investigated by extracellular matrix degradation assays and in vivo mouse model of metastasis.

Results

The data show that Shp1 was recruited to invadopodia and promoted the dephosphorylation of cortactin at tyrosine 421, leading to an attenuated capacity of melanoma cancer cells to degrade the extracellular matrix. Controls included the use of short interference RNA and catalytically-dead mutant that prevented the dephosphorylation of cortactin and hence the decrease the extracellular matrix degradation by melanoma cells. In addition, the phosphoinositide metabolite glycerophosphoinositol facilitated the localization of Shp1 at invadopodia hence promoting cortactin dephosphorylation. This impaired invadopodia function and tumor dissemination both in vitro and in an in vivo model of melanomas.

Conclusion

The main finding here reported is that cortactin is a specific substrate of the tyrosine phosphatase Shp1 and that its phosphorylation/dephosphorylation affects invadopodia formation and, as a consequence, the ability of melanoma cells to invade the extracellular matrix. Shp1 can thus be considered as a regulator of melanoma cell invasiveness and a potential target for antimetastatic drugs.
NIPU: a randomised, open-label, phase II study evaluating nivolumab and ipilimumab combined with UV1 vaccination as second line treatment in patients with malignant mesothelioma
Authors: Haakensen Vilde Drageset , Nowak Anna K. , Ellingsen Espen Basmo , Farooqi Saima Jamil , Bjaanæs Maria Moksnes , Horndalsveen Henrik , Mcculloch Tine , Grundberg Oscar , Cedres Susana M. , Helland Åslaug
Published: June 2021

Background

Malignant pleural mesothelioma (MPM) is a rare and aggressive tumour. For patients with inoperable disease, few treatment options are available after first line chemotherapy. The combination of ipilimumab and nivolumab has recently shown increased survival compared to standard chemotherapy, but most patients do not respond and improvements are called for. Telomerase is expressed in mesothelioma cells, but only sparsely in normal tissues and is therefore an attractive target for therapeutic vaccination. Vaccination against telomerase is tolerable and has shown to induce immune responses associated with increased survival in other cancer types. There is a well-founded scientific rationale for the combination of a telomerase vaccine and checkpoint inhibition to improve treatment response in MPM patients.

Methods

NIPU is a randomized, multi-centre, open-label, phase II study comparing the efficacy and safety of nivolumab and ipilimumab with or without telomerase vaccine in patients with inoperable malignant pleural mesothelioma after first-line platinum-based chemotherapy. Participants (n = 118) are randomized 1:1 into two treatment arms. All participants receive treatment with nivolumab (240 mg every 2 weeks) and ipilimumab (1 mg/kg every 6 weeks) until disease progression, unacceptable toxicity or for a maximum of 2 years. Patients randomised to the experimental arm receive 8 intradermal injections of UV1 vaccine during the first three months of treatment. Tumour tissue, blood, urine, faeces and imaging will be collected for biomarker analyses and exploration of mechanisms for response and resistance to therapy.

Discussion

Checkpoint inhibition is used for treatment of mesothelioma, but many patients still do not respond. Increasing therapy response to immunotherapy is an important goal. Possible approaches include combination with chemotherapy, radiotherapy, targeted therapy and other immunotherapeutic agents. Predictive biomarkers are necessary to ensure optimal treatment for each patient and to prevent unnecessary side effects. This trial seeks to improve treatment response by combining checkpoint inhibition with a telomerase vaccine and also to explore mechanisms for treatment response and resistance. Knowledge gained in the NIPU study may be transferred to the first line setting and to other cancers with limited benefit from immunotherapy.

Trial registration: ClinicalTrials.gov: NCT04300244, registered March 8th, 2020, https://clinicaltrials.gov/ct2/show/NCT04300244?term=NIPU&draw=2&rank=1.
The effect of revascularization on recovery of mitochondrial respiration in peripheral artery disease: a case control study
Authors: Gratl Alexandra , Pesta Dominik , Gruber Leonhard , Speichinger Fiona , Raude Ben , Omran Safwan , Greiner Andreas , Frese Jan Paul
Published: June 2021

Background

Peripheral arterial disease (PAD) is accompanied by myopathy characterized by mitochondrial dysfunction. The aim of this experimental study was to investigate the effect of revascularization procedures on mitochondrial function in ischemic and non-ischemic muscle.

Methods

Muscle biopsies from patients with symptomatic stage IIB/III PAD caused by isolated pathologies of the superficial femoral artery were obtained from muscle regions within the chronic ischemic muscle (gastrocnemius) and from non-ischemic muscle (vastus lateralis) before and 6 weeks after invasive revascularization. High-resolution respirometry was used to investigate mitochondrial function and results were normalized to citrate synthase activity (CSA). Results are given in absolute values and fold over basal (FOB).

Results

Respiratory states (OXPHOS (P) and electron transfer (E) capacity) normalized to CSA decreased while CSA was increased in chronic ischemic muscle after revascularization. There were no changes in in non-ischemic muscle. The FOB of chronic ischemic muscle was significantly higher for CSA (chronic ischemic 1.37 (IQR 1.10–1.64) vs. non-ischemic 0.93 (IQR 0.69–1.16) p = 0.020) and significantly lower for respiratory states normalized to CSA when compared to the non-ischemic muscle (P per CSA chronic ischemic 0.64 (IQR 0.46–0.82) vs non-ischemic 1.16 (IQR 0.77–1.54) p = 0.011; E per CSA chronic ischemic 0.61 (IQR 0.47–0.76) vs. non-ischemic 1.02 (IQR 0.64–1.40) p = 0.010).

Conclusions

Regeneration of mitochondrial content and function following revascularization procedures only occur in muscle regions affected by malperfusion. This indicates that the restoration of blood and oxygen supply are important mediators aiding mitochondrial recovery.
Necrotizing Funisitis as an Intrauterine manifestation of Cryopyrin-Associated Periodic Syndrome: a case report and review of the literature
Authors: Yokoi Kyoko , Minamiguchi Sachiko , Honda Yoshitaka , Kobayashi Mizuho , Kobayashi Satoru , Nishikomori Ryuta
Published: June 2021

Background

Cryopyrin-associated periodic syndrome (CAPS) is a life-long, autoinflammatory disease associated with a gain-of-function mutation in the nucleotide-binding domain, leucine-rich repeat family, pyrin domain containing 3 (NLRP3) gene, which result in uncontrolled production of IL-1β and chronic inflammation. Chronic infantile neurologic cutaneous and articular (CINCA) syndrome/neonatal-Onset multisystem inflammatory disease (NOMID) is the most severe form of CAPS. Although the first symptoms may be presented at birth, there are few reports on the involvement of the placenta and umbilical cord in the disease. Therefore, we present herein a preterm case of CINCA/NOMID syndrome and confirms intrauterine-onset inflammation with conclusive evidence by using fetal and placental histopathological examination.

Case presentation

The female patient was born at 33weeks of gestation by emergency caesarean section and weighted at 1,514 g. The most common manifestations of CINCA/NOMID syndrome including recurrent fever, urticarial rash, and ventriculomegaly due to aseptic meningitis were presented. She also exhibited atypical symptoms such as severe hepatosplenomegaly with cholestasis. The genetic analysis of NLRP3 revealed a heterozygous c.1698 C > G (p.Phe566Leu) mutation, and she was diagnosed with CINCA/NOMID syndrome. Further, a histopathological examination revealed necrotizing funisitis, mainly inflammation of the umbilical artery, along with focal neutrophilic and lymphocytic villitis.

Conclusions

The necrotizing funisitis, which only involved the artery, was an unusual observation for chorioamnionitis. These evidences suggest that foetal inflammation, probably due to overproduction of IL-1β, caused tissue damage in utero, and the first symptom of a newborn with CINCA/NOMID.
Impact of COVID-19 pandemic on HIV viremia: a single-center cohort study in northern Italy
Authors: Izzo Ilaria , Carriero Canio , Gardini Giulia , Fumarola Benedetta , Chiari Erika , Castelli Francesco , Quiros-Roldan Eugenia
Published: June 2021

Background

Brescia Province, northern Italy, was one of the worst epicenters of the COVID-19 pandemic. The division of infectious diseases of ASST (Azienda Socio Sanitaria Territoriale) Spedali Civili Hospital of Brescia had to face a great number of inpatients with severe COVID-19 infection and to ensure the continuum of care for almost 4000 outpatients with HIV infection actively followed by us. In a recent manuscript we described the impact of the pandemic on continuum of care in our HIV cohort expressed as number of missed visits, number of new HIV diagnosis, drop in ART (antiretroviral therapy) dispensation and number of hospitalized HIV patients due to SARS-CoV-2 infection. In this short communication, we completed the previous article with data of HIV plasmatic viremia of the same cohort before and during pandemic.

Methods

We considered all HIV-patients in stable ART for at least 6 months and with at least 1 available HIV viremia in the time window March 01–November 30, 2019, and another group of HIV patients with the same two requisites but in different time windows of the COVID-19 period (March 01–May 31, 2020, and June 01–November 30, 2020). For patients with positive viremia (PV) during COVID-19 period, we reported also the values of viral load (VL) just before and after PV. Results: the percentage of patients with PV during COVID-19 period was lower than the previous year (2.8% vs 7%). Only 1% of our outpatients surely suffered from pandemic in term of loss of previous viral suppression.

Conclusions

Our efforts to limit the impact of pandemic on our HIV outpatients were effective to ensure HIV continuum of care.
Lymphocyte–monocyte–neutrophil index: a predictor of severity of coronavirus disease 2019 patients produced by sparse principal component analysis
Authors: Qi Yingjie , Jia Jian-an , Li Huiming , Wan Nagen , Zhang Shuqin , Ma Xiaoling
Published: June 2021

Background

It is important to recognize the coronavirus disease 2019 (COVID-19) patients in severe conditions from moderate ones, thus more effective predictors should be developed.

Methods

Clinical indicators of COVID-19 patients from two independent cohorts (Training data: Hefei Cohort, 82 patients; Validation data: Nanchang Cohort, 169 patients) were retrospected. Sparse principal component analysis (SPCA) using Hefei Cohort was performed and prediction models were deduced. Prediction results were evaluated by receiver operator characteristic curve and decision curve analysis (DCA) in above two cohorts.

Results

SPCA using Hefei Cohort revealed that the first 13 principal components (PCs) account for 80.8% of the total variance of original data. The PC1 and PC12 were significantly associated with disease severity with odds ratio of 4.049 and 3.318, respectively. They were used to construct prediction model, named Model-A. In disease severity prediction, Model-A gave the best prediction efficiency with area under curve (AUC) of 0.867 and 0.835 in Hefei and Nanchang Cohort, respectively. Model-A’s simplified version, named as LMN index, gave comparable prediction efficiency as classical clinical markers with AUC of 0.837 and 0.800 in training and validation cohort, respectively. According to DCA, Model-A gave slightly better performance than others and LMN index showed similar performance as albumin or neutrophil-to-lymphocyte ratio.

Conclusions

Prediction models produced by SPCA showed robust disease severity prediction efficiency for COVID-19 patients and have the potential for clinical application.
Lithium inhibits NF-κB nuclear translocation and modulate inflammation profiles in Rift valley fever virus-infected Raw 264.7 macrophages
Authors: Makola Raymond Tshepiso , Kgaladi Joe , More Garland Kgosi , Jansen van Vuren Petrus , Paweska Janusz Tadeusz , Matsebatlela Thabe Moses
Published: June 2021

Introduction

Rift Valley fever virus (RVFV) is a zoonotic life-threatening viral infection endemic across sub-Saharan African countries and the Arabian Peninsula; however, there is a growing panic of its spread to non-endemic regions. This viral infection triggers a wide spectrum of symptoms that span from fibril illnesses to more severe symptoms such as haemorrhagic fever and encephalitis. These severe symptoms have been associated with dysregulated immune response propagated by the virulence factor, non-structural protein (NSs). Thus, this study investigates the effects of lithium on NF-κB translocation and RFVF-induced inflammation in Raw 264.7 macrophages.

Methods

The supernatant from RVFV-infected Raw 264.7 cells, treated with lithium, was examined using an ELISA assay kit to measure levels of cytokines and chemokines. The H2DCF-DA and DAF-2 DA florigenic assays were used to determine the levels of ROS and RNS by measuring the cellular fluorescence intensity post RVFV-infection and lithium treatment. Western blot and immunocytochemistry assays were used to measure expression levels of the inflammatory proteins and cellular location of the NF-κB, respectively.

Results

Lithium was shown to stimulate interferon-gamma (IFN-γ) production as early as 3 h pi. Production of the secondary pro-inflammatory cytokine and chemokine, interleukin-6 (IL-6) and regulated on activation, normal T cell expressed and secreted (RANTES), were elevated as early as 12 h pi. Treatment with lithium stimulated increase of production of tumor necrosis factor-alpha (TNF-α) and Interleukin-10 (IL-10) in RVFV-infected and uninfected macrophages as early as 3 h pi. The RVFV-infected cells treated with lithium displayed lower ROS and RNS production as opposed to lithium-free RVFV-infected control cells. Western blot analyses demonstrated that lithium inhibited iNOS expression while stimulating expression of heme oxygenase (HO) and IκB in RVFV-infected Raw 264.7 macrophages. Results from immunocytochemistry and Western blot assays revealed that lithium inhibits NF-κB nuclear translocation in RVFV-infected cells compared to lithium-free RVFV-infected cells and 5 mg/mL LPS controls.

Conclusion

This study demonstrates that lithium inhibits NF-kB nuclear translocation and modulate inflammation profiles in RVFV-infected Raw 264.7 macrophage cells.