bluuebunny/crossref_metadata_2025 huggingface dataset:
A novel genomic panel as an adjunctive diagnostic tool for the characterization and profiling of breast Fibroepithelial lesions
Authors: Sim Yirong , Ng Gwendolene Xin Pei , Ng Cedric Chuan Young , Rajasegaran Vikneswari , Wong Suet Far , Liu Wei , Guan Peiyong , Nagarajan Sanjanaa , Ng Wai Yee , Thike Aye Aye , Lim Jeffrey Chun Tatt , Nasir Nur Diyana Binte Md , Tan Veronique Kiak Mien , Madhukumar Preetha , Yong Wei Sean , Wong Chow Yin , Tan Benita Kiat Tee , Ong Kong Wee , Teh Bin Tean , Tan Puay Hoon
Published: October 2019

Background

Known collectively as breast fibroepithelial lesions (FELs), the common fibroadenomas (FAs) and the rarer phyllodes tumors (PTs) are a heterogenous group of biphasic neoplasms. Owing to limited tissue availability, inter-observer variability, overlapping histological features and heterogeneity of these lesions, diagnosing them accurately on core biopsies is challenging. As the choice management option depends on the histological diagnosis; a novel 16-gene panel assay was developed to improve the accuracy of preoperative diagnosis on core biopsy specimens.

Methods

Using this 16-gene panel, targeted amplicon-based sequencing was performed on 275 formalin-fixed, paraffin-embedded (FFPE) breast FEL specimens, archived at the Singapore General Hospital, from 2008 to 2012.

Results

In total, 167 FAs, 24 benign, 14 borderline and 6 malignant PTs, were profiled. Compared to FAs, PTs had significantly higher mutation rates in theTERTpromoter (p <  0.001),RARA(p <  0.001),FLNA,RB1andTP53(p = 0.002, 0.020 and 0.018, respectively). In addition to a higher mutational count (p <  0.001),TERTpromoter (p <  0.001), frameshift, nonsense and splice site (p = 0.001, < 0.001 and 0.043, respectively) mutations were also frequently observed in PTs.

A multivariate logistic regression model was built using these as variables and a predictive scoring system was developed. It classifies a FEL at low or high risk (score <  1 and ≥ 1, respectively) of being a PT. This scoring system has good discrimination (ROC area = 0.773, 95% CI: 0.70 to 0.85), calibration (p = 0.945) and is significant in predicting PTs (p <  0.001).

Conclusion

This novel study demonstrates the ability to extract DNA of sufficient quality and quantity for targeted sequencing from FFPE breast core biopsy specimens, along with their successful characterization and profiling using our customized 16-gene panel. Prospective work includes validating the utility of this promising 16-gene panel assay as an adjunctive diagnostic tool in clinical practice.
Characterization of a highly stable α-galactosidase from thermophilic Rasamsonia emersonii heterologously expressed in a modified Pichia pastoris expression system
Authors: An Jian-Lu , Zhang Wei-Xin , Wu Wei-Ping , Chen Guan-Jun , Liu Wei-Feng
Published: October 2019

Background

Structurally stable α-galactosidases are of great interest for various biotechnological applications. More thermophilic α-galactosidases with high activity and structural stability have therefore to be mined and characterized. On the other hand, few studies have been performed to prominently enhance the AOX1 promoter activity in the commonly used Pichia pastoris system, in which production of some heterologous proteins are insufficient for further study.

Results

ReGal2 encoding a thermoactive α-galactosidase was identified from the thermophilic (hemi)cellulolytic fungus Rasamsonia emersonii. Significantly increased production of ReGal2 was achieved when ReGal2 was expressed in an engineered Pastoris pichia expression system with a modified AOX1 promoter and simultaneous fortified expression of Mxr1 that is involved in transcriptionally activating AOX1. Purified ReGal2 exists as an oligomer and has remarkable thermo-activity and thermo-tolerance, exhibiting maximum activity of 935 U/mg towards pNPGal at 80 °C and retaining full activity after incubation at 70 °C for 60 h. ReGal2 is insensitive to treatments by many metal ions and exhibits superior tolerance to protein denaturants. Moreover, ReGal2 efficiently hydrolyzed stachyose and raffinose in soybeans at 70 °C in 3 h and 24 h, respectively.

Conclusion

A modified P. pichia expression system with significantly enhanced AOX1 promoter activity has been established, in which ReGal2 production is markedly elevated to facilitate downstream purification and characterization. Purified ReGal2 exhibited prominent features in thermostability, catalytic activity, and resistance to protein denaturants. ReGal2 thus holds great potential in relevant biotechnological applications.
Evaluating the psychometric properties of the mental health continuum-short form (MHC-SF) in Dutch adolescents
Authors: Luijten Chantie C. , Kuppens Sofie , van de Bongardt Daphne , Nieboer Anna P.
Published: October 2019

Background

Mental health is increasingly viewed as the presence of various aspects of well-being rather than just the absence of mental illness. The Mental Health Continuum-Short Form (MHC-SF) is a 14-item instrument that assesses mental health, focusing on emotional, psychological, and social well-being. The present study examined for the first time the psychometric properties of the Dutch version of the MHC-SF among adolescents, focusing on its factor structure, internal consistency, construct validity, and gender and age factorial invariance.

Methods

Data were collected from a school-based sample of 1175 adolescents (53.4% girls) aged 11–17 years (M = 13.7; SD = 1.1). Participants completed an online questionnaire in the classroom during regular school hours. Statistical analyses to evaluate the factor structure, internal consistency, construct validity, and gender and age factorial invariance were performed in SPSS and R.

Results

Using confirmatory factor analyses, a satisfactory-to-good fit was obtained for the three-factor model (emotional, psychological, and social well-being). The MHC-SF scores showed good internal consistency (Cronbach’s alpha = .91) and results supported convergent and divergent validity. Finally, the MHC-SF showed gender and age factorial invariance.

Conclusion

The current psychometric evaluation indicates the MHC-SF is a reliable and valid instrument to assess multiple dimensions of well-being among Dutch adolescents. The instrument can be applied for research purposes and in clinical practice.
Notch pathway: a bistable inducer of biological noise?
Authors: Vujovic Filip , Hunter Neil , Farahani Ramin M.
Published: October 2019

Notch signalling pathway is central to development of metazoans. The pathway codes a binary fate switch. Upon activation, downstream signals contribute to resolution of fate dichotomies such as proliferation/differentiation or sub-lineage differentiation outcome. There is, however, an interesting paradox in the Notch signalling pathway. Despite remarkable predictability of fate outcomes instructed by the Notch pathway, the associated transcriptome is versatile and plastic. This inconsistency suggests the presence of an interface that compiles input from the plastic transcriptome of the Notch pathway but communicates only a binary output in biological decisions. Herein, we address the interface that determines fate outcomes. We provide an alternative hypothesis for the Notch pathway as a biological master switch that operates by induction of genetic noise and bistability in order to facilitate resolution of dichotomous fate outcomes in development.

Graphical abstract
A comparison of sexual desire in opiate-dependent men receiving methadone and buprenorphine maintenance treatment
Authors: Yee Anne , Loh Huai Seng , Loh Huai Heng , Riahi Shahrzad , Ng Chong Guan , Sulaiman Ahmad Hatim bin
Published: October 2019

Background

Methadone is an effective therapy for opiate dependence. However, one of the commonest side effects is sexual dysfunction among male patients. Buprenorphine is an alternative to methadone. This study aimed to compare sexual desire among opiate-dependent male patients on buprenorphine (BMT) and methadone maintenance therapy (MMT).

Methods

This cross-sectional study involved 126 male opiate-dependent patient who were tested for total testosterone (TT) and prolactin levels, and were interviewed and completed the Sexual Desire Inventory-2 (SDI-2), Malay language of International Index of Erectile Function (Mal-IIEF-15) and the Malay version of the self-rated Montgomery-Asberg Depression Rating Scale (MADRS-BM) questionnaires.

Results

There were 95 (75.4%) patients on MMT and 31 (24.6%) on BMT. Patients on MMT scored significantly lower in the sexual desire domain (Mal-IIEF-15 scores) (p < 0.01), dyadic sexual desire (p = 0.04) and TT plasma level (p < 0.01) when compared to BMT group after controlling all the confounders.

Conclusions

Patients on MMT are associated with lower sexual desire when compared with patients on BMT. Smoking may further lower testosterone and, hence, sexual desire in those already on methadone.
Clinicopathologic and microenvironmental analysis of primary cutaneous CD30-positive lymphoproliferative disorders: a 26 year experience from an academic medical center in Brazil
Authors: Ferreira Cristiane Rúbia , Zhao Shuchun , Sanches José Antonio , Miyashiro Denis , Cury-Martins Jade , Azevedo Raymundo Soares , Zerbini Maria C. N. , Natkunam Yasodha , Gratzinger Dita
Published: October 2019

Background

Primary cutaneous CD30+ lymphoproliferative disorders (pc-CD30-LPD) are a group of clonal T cell lymphoproliferative disorders that despite very similar tumor histology follow different and characteristic clinical courses, suggesting a homeostatic role of the tumor microenvironment. Little is known about tumor microenvironment and there is almost no literature about PD-L1 expression in pc-CD30-LPD.

Methods

This retrospective study presents a fully clinicopathologically characterized series of pc-CD30-LPDs from an academic medical center in Brazil, including 8 lymphomatoid papulomatosis (LyP), 9 primary cutaneous anaplastic large cell lymphoma (pcALCL) and 4 borderline lesions. All the cases were scored for FOXP3+ regulatory T-cells (Treg) and CD8+ cytotoxic tumor infiltrating lymphocytes (TIL) densities, as well as PD-L1 expression in tumor cells and tissue associated macrophages. The CD8+/FOXP3+ ratio was also evaluated.

Results

Among the 21 cases of pc-CD30-LPD, PD-L1 expression is frequent in both tumor cells and tissue associated macrophages in pc-CD30-LPD across categories, suggesting that the PD-L1 axis may be a common feature of pc-CD30-LPDs. While reactive T cell infiltrates vary widely from case to case, a common feature across pc-CD30-LPDs is higher density of CD8 than FOXP3 + T cells. The distribution of T cells within the lesions however differed between LyP and pcALCL: we found that LyP lesions tend to be permeated by CD8+ and FOXP3+ T cells, whereas pcALCL tend to be surrounded by a rim of CD8+ TIL and FOXP3+ Tregs with relatively lower density infiltrates in the center of the lesion.

Conclusions

LyP has a trend to have denser immune cells throughout the lesion, with higher FOXP3+ Treg and CD8+ TIL in the center than the edge comparing with pcALCL. PD-L1+ is frequent in tumor cells and tissue associated macrophages in pc-CD30-LPD. The differential distribution of CD8+ and FOXP3+ TILs in LyP as compared to pcALCL could provide a clue to the relapsing/remitting course of LyP as compared to the less frequent spontaneous regression of pcALCL.
Assessing plant performance in the Enviratron
Authors: Bao Yin , Zarecor Scott , Shah Dylan , Tuel Taylor , Campbell Darwin A. , Chapman Antony V. E. , Imberti David , Kiekhaefer Daniel , Imberti Henry , Lübberstedt Thomas , Yin Yanhai , Nettleton Dan , Lawrence-Dill Carolyn J. , Whitham Steven A. , Tang Lie , Howell Stephen H.
Published: October 2019

Background

Assessing the impact of the environment on plant performance requires growing plants under controlled environmental conditions. Plant phenotypes are a product of genotype × environment (G × E), and the Enviratron at Iowa State University is a facility for testing under controlled conditions the effects of the environment on plant growth and development. Crop plants (including maize) can be grown to maturity in the Enviratron, and the performance of plants under different environmental conditions can be monitored 24 h per day, 7 days per week throughout the growth cycle.

Results

The Enviratron is an array of custom-designed plant growth chambers that simulate different environmental conditions coupled with precise sensor-based phenotypic measurements carried out by a robotic rover. The rover has workflow instructions to periodically visit plants growing in the different chambers where it measures various growth and physiological parameters. The rover consists of an unmanned ground vehicle, an industrial robotic arm and an array of sensors including RGB, visible and near infrared (VNIR) hyperspectral, thermal, and time-of-flight (ToF) cameras, laser profilometer and pulse-amplitude modulated (PAM) fluorometer. The sensors are autonomously positioned for detecting leaves in the plant canopy, collecting various physiological measurements based on computer vision algorithms and planning motion via “eye-in-hand” movement control of the robotic arm. In particular, the automated leaf probing function that allows the precise placement of sensor probes on leaf surfaces presents a unique advantage of the Enviratron system over other types of plant phenotyping systems.

Conclusions

The Enviratron offers a new level of control over plant growth parameters and optimizes positioning and timing of sensor-based phenotypic measurements. Plant phenotypes in the Enviratron are measured in situ—in that the rover takes sensors to the plants rather than moving plants to the sensors.
Critique of the concept of motivation and its implications for healthcare practices
Authors: Sampaio Leonardo Augusto Negreiros Parente , de Carvalho Mesquita Ayres José Ricardo
Published: October 2019

Background

Motivation is a crucial and widespread theme within medicine. From clinical to surgical scenarios, acquiescence in taking a pill or coming to a consultation is imperative for medical treatment to thrive. The “decade of the brain” gave practitioners substantial neuroscientific data on human behavior, helped to explain why people do what they do and created the concept of “motivated brain”. Findings from empirical psychology stratified motivation into stages of change, which became more complex over the decades. This research seeks to improve the understanding of how people make decisions about their health, and how to better understand strategies and techniques to help them resolve ambivalence in an effective goal-oriented way.

Methods

We establish a dialogue with Ricoeur’s phenomenology of the will in order to understand the meaning of these scientific findings. Starting from Husserlian phenomenology, Paul Ricoeur developed his thoughts away from transcendental idealism, through emancipating the intentional structures of the will from the realm of perception.

Results

Through introducing the concepts of the voluntary and the involuntary, Ricoeur deviated from Cartesian dualism, which renders the body as an object body, a target of natural vicissitudes. The new dualism of the voluntary and the involuntary is dealt with by reference to what Ricoeur called the central mystery of incarnate existence, which considers man “double inhumanity, simple invitality”. This duality makes it possible to consider the brain to be the natural organ of behavior in the human body, and to use empirical psychology as a path to escape from shallow subjectivations of concepts.

Conclusions

Paul Ricoeur’s simplicity (or unity) of existence provides an invitation for medicine to rethink some of its philosophical assumptions, such that patients can be considered to be autonomous subjects with authorial life projects. Ricoeurian anthropology has a deep ethical impact on how medicine should use technology, which arises from empirical psychology findings. The usage of this new knowledge also needs to be thoroughly inspected, since it shifts the social role of medical science.
Association between cam-type femoroacetabular impingement and osteitis pubis in non-athletic population on magnetic resonance imaging
Authors: Akgün Ayşe Serap , Agirman Mehmet
Published: October 2019

Background

Osteitis pubis (OP) is a common source of groin and extra-articular hip pain and is associated with intra-articular hip pathology. In this study, we aimed to determine the prevalence of osteitis pubis on magnetic resonance imaging (MRI) in non-athletic patients with cam-type femoroacetabular impingement (FAI).

Methods

This retrospective cross-sectional study included 178 subjects: 90 patients with cam-type FAI diagnosed by MRI and 88 subjects used as a control group. Additionally, their MRI data were analyzed for the characteristics of osteitis pubis, with severity graded from minimal to severe on a four-point scale.

Results

A total of 98 patients and 88 controls were studied. Seventy-two males (80%) and 18 females (20%) were the patient group, whereas 71 males (80.68%) and 17 females (19.32%) were the control group. The mean alpha angle of the patients with FAI was 65.8 ± 3.3° in the right side and 66.2 ± 3.2° in the left side, whereas in the control group, it was 47 ± 5.6° in the right side and 47.8 ± 5.2° in the left side. Alpha angle measurements were significantly higher in the patient group than the control group (p < 0.001). A statistically significant increase in the prevalence of osteitis pubis was found in patients with cam-type FAI (45.56%) compared to control subjects (5.68%) (p < 0.001).

Conclusions

This study demonstrated that the frequency of osteitis pubis was increased in non-athletic patients with FAI syndrome. Further studies are required to determine whether these findings reflect the clinical symptoms in patients with hip pain.
Clinical results and surgery tactics of spinal osteotomy for ankylosing spondylitis kyphosis: experience of 428 patients
Authors: Xin Zhijun , Zheng Guoquan , Huang Peng , Zhang Xuesong , Wang Yan
Published: October 2019

Objective

To report the clinical results and surgical tactics of spinal osteotomy for ankylosing spondylitis (AS) kyphosis based on the experiences of 428 patients.

Methods

From January 2003 to January 2015, a total of 428 patients suffering from AS kyphosis who underwent a one- or two-level pedicle subtraction osteotomy (PSO) or vertebral column decancellation (VCD) osteotomy in our hospital were reviewed. Pre- and postoperative radiological parameters and the chin-brow vertical angle (CBVA) were measured. Intraoperative, postoperative, and general complications were recorded.

Results

All patients could walk with horizontal vision and lie on their backs postoperatively. The pre- and postoperative average global kyphosis (GK) angles were corrected from 82.6 to 12.7° (p = 0.000) in the two-level group and from 55.8 to 9.6° (p = 0.000) in the one-level group, respectively. The mean sagittal vertical axis (SVA) improved from 29.4 to 8 cm (p = 0.000) in the two-level group and from 18.0 to 4.3 cm (p = 0.000) in the one-level group. The CBVA improved from 68.3 to 8.2° (p = 0.000) in the two-level group and from 46.2 to 4.2° (p = 0.000) in the one-level group. Although no major acute complications such as death or complete paralysis occurred, the complication rate was 6.5% in the one-level group and 23.6% in the two-level group.

Conclusion

Spinal osteotomy, such as PSO and VCD, can improve the quality of life of AS patients as well as correct kyphotic deformities. The one-level spinal osteotomy showed a lower complication rate, while two-level spinal osteotomy was a relatively aggressive procedure that was more suitable in correcting severe AS kyphotic deformities.
Influence of bone density on morphologic cement penetration in minimally invasive tibial unicompartmental knee arthroplasty: an in vitro cadaver study
Authors: Scheele Christian B. , Pietschmann Matthias F. , Schröder Christian , Lazic Igor , Grupp Thomas M. , Müller Peter E.
Published: October 2019

Background

Unicompartmental knee arthroplasty is an established treatment option for anteromedial osteoarthritis. However, large registry studies report higher rates of aseptic loosening compared to total knee arthroplasty. The objective of this study was to assess the impact of bone density on morphological cement penetration. Moreover, an alternative regional bone density measuring technique was validated against the established bone mineral density assessment.

Methods

Components were implanted on the medial side of 18 fresh-frozen cadaver knees using a minimally invasive approach. Bone density has been quantified prior to implantation using Hounsfield units and bone mineral density. Morphological cement penetration has been assessed in different areas and was correlated with local bone density.

Findings

A highly significant correlation between Hounsfield units and trabecular bone mineral density was detected (r = 0.93;P < 0.0001), and local bone density was significantly increased in the anterior and posterior area (P = 0.0003). The mean cement penetration depth was 1.5 (SD 0.5 mm), and cement intrusion into trabecular bone was interrupted in 31.8% (SD 23.7%) of the bone-cement interface. Bone density was correlated significantly negative with penetration depth (r = − 0.31;P = 0.023) and positive with interruptions of horizontal interdigitating (r = + 0.33;P = 0.014). Cement penetration around the anchoring peg was not significantly correlated with bone density.

Interpretation

Areas with high bone density were characterized by significantly lower penetration depths and significantly higher areas without cement penetration. Anchoring pegs facilitate cement intrusion mechanically. Regional quantification of bone density using Hounsfield units is a simple but valuable extension to the established determination of bone mineral density.
The risk factors for postoperative cerebral complications in patients with Stanford type a aortic dissection
Authors: Lin Yong , Chen Mei-Fang , Zhang Hui , Li Ruo-Meng , Chen Liang-Wan
Published: October 2019

Background

Postoperative cerebral complications (PCC) are common and serious postoperative complications for patients with Stanford type A aortic dissection (AAD). The aim of this study was to evaluate the risk factors for PCC in these patients and to provide a scientific basis for effective prevention of PCC.

Methods

In this retrospective case-control study, 125 patients with AAD who underwent thoracotomy in our department from October 2017 to October 2018 in the department of cardiovascular surgery, Fujian Medical University Union Hospital were divided into two groups: patients with PCC (n = 12), and patients without PCC (n = 113). The general clinical data, the types of corrective surgeries, the intraoperative situations, the postoperative complications, and the midterm outcomes of the patients were analyzed.

Results

The patients with PCC were significantly older than the patients without PCC (P = 0.016), and the incidence of the preoperative cerebral disease history in the patients with PCC was significantly higher than those of the PCC (−) group (P = 0.024). The Euro SCORE II of patients with PCC was dramatically higher than the patients without PCC (P = 0.005). There were significant differences between the two groups in terms of the duration of cardiopulmonary bypass (CPB) (P = 0.010) and the length of moderate hypothermic circulatory arrest (MHCA) combined with selective cerebral perfusion (SCP) (P = 0.000). The monitoring of rcSO2 indicated that there was significant difference between the two groups in terms of the bilateral baseline (P = 0.000). Patients with PCC were observed to have experienced significantly longer intubation times (P = 0.000), ICU stays (P = 0.001), and postoperative hospital stays (P = 0.009), and they also had dramatically higher rates of pulmonary infection (P = 0.000), multiple organ dysfunction syndrome (P = 0.041) and tracheotomy (P = 0.022) after surgeries. The duration of MHCA+SCP (OR:9.009, P = 0.034) and the average baseline value of rcSO2 (OR:0.080, P = 0.009) were ultimately identified as significant risk factors.

Conclusions

PCC has a serious influence on the prognoses of patients following surgical treatment with AAD. The duration of MHCA+SCP and the average baseline value of rcSO2 were the independent risk factors for PCC.
A placebo-controlled trial of folic acid and betaine in identical twins with Angelman syndrome
Authors: Han Julia , Bichell Terry Jo , Golden Stephanie , Anselm Irina , Waisbren Susan , Bacino Carlos A. , Peters Sarika U. , Bird Lynne M. , Kimonis Virginia
Published: October 2019

Background

Angelman syndrome (AS) is a neurodevelopmental disorder that is caused by maternal genetic deficiency of a gene that encodes E6-AP ubiquitin-protein ligase (gene symbol UBE3A) mapping to chromosome 15q11-q13. AS leads to stiff and jerky gait, excess laughter, seizures, and severe intellectual disability. In some parts of the brain, the paternally inherited UBE3A gene is subject to genomic imprinting by the action of the UBE3A-antisense transcript (UBE3A-ATS) on the paternally inherited allele. Consequently, only the maternally inherited UBE3A gene is expressed in mature neurons. AS occurs due to deletions of the maternal 15q11 − 13 region, paternal uniparental disomy (UPD), imprinting center defects, mutations in the maternal UBE3A gene, or other unknown genetic malfunctions that result in a silenced maternal UBE3A gene in the specific imprinted regions of the brain.

Results

A potential treatment strategy for AS is to increase methylation of UBE3A-ATS to promote expression of the paternal UBE3A gene and thus ameliorate the clinical phenotypes of AS. We treated two sets of male identical twins with class I deletions with a 1 year treatment trial of either betaine and folic acid versus placebo. We found no statistically significant changes in the clinical parameters tested at the end of the 1 year trial, nor did we find any significant adverse events.

Conclusions

This study tested the hypothesis that by increasing the methylation of the UBE3A-antisense transcript in Angelman syndrome to promote expression of the silenced paternal UBE3A gene we may ameliorate the clinical phenotypes of AS. We treated two sets of identical twins with placebo versus betaine and folic acid. Although this study represented a novel approach to treating Angelman syndrome, the differences in the developmental testing results was not significant. This paper also discusses the value of monozygotic twin studies in minimizing confounding variables and its utility in conducting small treatment studies.

Trial registration

NCT00348933. Registered 6 July 2006.
Targeting tauopathy with engineered tau-degrading intrabodies
Authors: Gallardo Gilbert , Wong Connie H. , Ricardez Sara M. , Mann Carolyn N. , Lin Kent H. , Leyns Cheryl E. G. , Jiang Hong , Holtzman David M.
Published: October 2019

Background

The accumulation of pathological tau is the main component of neurofibrillary tangles and other tau aggregates in several neurodegenerative diseases, referred to as tauopathies. Recently, immunotherapeutic approaches targeting tau have been demonstrated to be beneficial in decreasing tauopathy in animal models. We previously found that passive immunotherapy with anti-tau antibody to human tau or expression of an anti-tau secreted single-chain variable fragment (scFv) in the central nervous system of a mouse model of tauopathy decreased but did not remove all tau-associated pathology. Although these and other studies demonstrate that conventional immunotherapeutic approaches targeting tau can influence tau pathogenesis, the majority of pathological tau remains in the cytosol of cells, not typically accessible to an extracellular antibody. Therefore, we reasoned targeting intracellular tau might be more efficacious in preventing or decreasing tauopathy.

Methods

By utilizing our anti-tau scFv, we generated anti-tau intrabodies for the expression in the cytosol of neurons. To enhance the degradation capacity of conventional intrabodies, we engineered chimeric anti-tau intrabodies fused to ubiquitin harboring distinct mutations that shuttle intracellular tau for either the proteasome or lysosomal mediated degradation. To evaluate the efficacy in delaying or eliminating tauopathy, we expressed our tau degrading intrabodies or controls in human tau transgenic mice by adeno-associated virus prior to overt tau pathology and after tau deposition.

Results

Our results demonstrate, the expression of chimeric anti-tau intrabodies significantly reduce tau protein levels in primary neuronal cultures expression human tau relative to a non-modified anti-tau intrabody. We found the expression of engineered tau-degrading intrabodies destined for proteasomal-mediated degradation are more effective in delaying or eliminating tauopathy than a conventional intrabody in aged human tau transgenic mice.

Conclusion

This study, harnesses the strength of intrabodies that are amendable for targeting specific domains or modifications with the cell-intrinsic mechanisms that regulate protein degradation providing a new immunotherapeutic approach with potentially improved efficacy.
Demographic and clinical characteristics of children seeking psychiatric services in the Nile Delta region: an observational retrospective study
Authors: Seleem Mohammad A. , Amer Reham A. , Romeh Amr H. , Hamoda Hesham M.
Published: October 2019

Background

Epidemiological studies, describing both community and clinical samples of youth in need for psychiatric help, are rare in the middle east. To our knowledge, this is the first study that aims to investigate the demographic and clinical characteristics of a sample of children suffering from emotional and behavioral problems seeking psychiatric services in the Nile Delta region and the largest clinical sample to date in Egypt.

Methods

The files of all new cases who presented for care in the outpatient service for children and adolescents between August 2016 and July 2018 were reviewed. Ninety-six files were excluded due to missing data while another 18 files were found to be for adults (ages > 18 years old), so the sample included 886 cases.

Results

The ages of our sample (n = 886) ranged from 18 months to 18 years with an average of 7.5 (± 3.8) years. Most of our cases were male, school aged children, living within low-income households and predominantly coming from rural areas. The most common diagnoses were attention deficit hyperactivity disorder (ADHD) (22.6%), intellectual disability (ID) (13.7%), depressive disorders (13.3%), and disruptive behavior disorders (DBD) (12.3%). Strong protective effects were found for family integrity and stability. Corporal punishment and physical and sexual abuse were noted to be significant risk factors for internalizing and externalizing disorders in children and adolescents.

Conclusions

Except for males being a majority in our sample of children seeking psychiatric consultation, demographic patterns and prevalence of psychiatric disorders are comparable to other tertiary clinical samples in other parts of the world.
Executive functioning and neurodevelopmental disorders in early childhood: a prospective population-based study
Authors: Otterman D. Louise , Koopman-Verhoeff M. Elisabeth , White Tonya J. , Tiemeier Henning , Bolhuis Koen , Jansen Pauline W.
Published: October 2019

Background

Executive functioning deficits are common in children with neurodevelopmental disorders. However, prior research mainly focused on clinical populations employing cross-sectional designs, impeding conclusions on temporal neurodevelopmental pathways. Here, we examined the prospective association of executive functioning with subsequent autism spectrum disorder (ASD) traits and attention-deficit/hyperactivity disorder (ADHD) traits.

Methods

This study included young children from the Generation R Study, a general population birth cohort. The Brief Rating Inventory of Executive Function-Preschool Version was used to assess parent-reported behavioral executive functioning when the children were 4 years old. ASD traits were assessed at age 6 (n = 3938) using the parent-reported Social Responsiveness Scale. The Teacher Report Form was used to assess ADHD traits at age 7 (n = 2749). Children with high scores were screened to determine possible clinical ASD or ADHD diagnoses. We were able to confirm an ASD diagnosis for n = 56 children by retrieving their medical records and established an ADHD diagnosis for n = 194 children using the Diagnostic Interview Schedule for Children-Young Child version (DISC-YC). Data were analyzed using hierarchical linear and logistic regressions.

Results

Impaired executive functioning was associated with more ASD and ADHD traits across informants (for ASD traits and diagnoses: β = 0.33, 95% CI [0.30–0.37]; OR = 2.69, 95% CI [1.92–3.77], respectively; for ADHD traits and diagnoses: β = 0.12, 95% CI [0.07–0.16]; OR = 2.32, 95% CI [1.89–2.85], respectively). Deficits in all subdomains were associated with higher levels of ASD traits, whereas only impaired inhibition, working memory, and planning/organization were associated with more ADHD traits.

Conclusions

The findings of the current study suggest a graded association of executive functioning difficulties along the continuum of ASD and ADHD and that problems in executive functioning may be a precursor of ASD and ADHD traits from an early age onwards.
Process development for pandemic influenza VLP vaccine production using a baculovirus expression system
Authors: Lai Chia-Chun , Cheng Yu-Chieh , Chen Pin-Wen , Lin Ting-Hui , Tzeng Tsai-Teng , Lu Chia-Chun , Lee Min-Shi , Hu Alan Yung-Chih
Published: October 2019

Background

Influenza viruses cause hundreds of thousands of respiratory diseases worldwide each year, and vaccination is considered the most effective approach for preventing influenza annual epidemics or pandemics. Since 1950, chicken embryonated eggs have been used as the main method for producing seasonal influenza vaccines. However, this platform has the main drawback of a lack of scale-up flexibility, and thus, egg-based vaccine manufacturers cannot supply sufficient doses within a short period for use for pandemic prevention. As a result, strategies for reducing the manufacturing time and increasing production capacity are urgently needed. Non-virion vaccine methods have been considered an alternative strategy against an influenza pandemic, and the purpose of maintaining an immunogenic capsule structure with infectious properties appears to be met by the virus-like particle (VLP) platform.

Results

An influenza H7N9-TW VLP production platform using insect cells, which included the expression of hemagglutinin (HA), NA, and M1 proteins, was established. To scale up H7N9-TW VLP production, several culture conditions were optimized to obtain a higher production yield. A high level of dissolved oxygen (DO) could be critical to H7N9-TW VLP production. If the DO was maintained at a high level, the HA titer obtained in the spinner flask system with ventilation was similar to that obtained in a shake flask. In this study, the HA titer in a 5-L bioreactor with a well-controlled DO level was substantially improved by 128-fold (from 4 HA units (HAU)/50 μL to 512 HAU/50 μL).

Conclusions

In this study, a multigene expression platform and an effective upstream process were developed. Notably, a high H7N9-TW VLP yield was achieved using a two-step production strategy while a high DO level was maintained. The upstream process, which resulted in high VLP titers, could be further used for large-scale influenza VLP vaccine production.
The high-affinity IgG receptor FcγRI modulates peripheral nerve injury-induced neuropathic pain in rats
Authors: Liang Yingxia , Zhang Zhiyu , Juan Zhaodong , Zhang Rui , Zhang Can
Published: October 2019

The Fc gamma receptor I (FcγRI; CD64) is the high-affinity receptor of the immunoglobulin G protein (IgG). It is usually expressed in immune cells and has recently been identified to distribute in the nervous system and play critical roles in various neurological disorders. Presently, the impacts of FcγRI in neuropathic pain was largely unknown. Here, we aimed to investigate the impacts of FcγRI in neuropathic pain through pain-related neurobehavioral studies and underlying mechanisms by biochemical methods in animal and cell models. Specifically, we first utilized the chronic constriction injury (CCI) rat model that displayed neuropathic pain related symptoms and signs, including thermal hyperalgesia and mechanical allodynia. These neurobehavioral defects were significantly attenuated by the anti-FcγRI antibody, which was associated with reduced levels of neuropeptide substance P, C3, and TNF-α. Furthermore, we validated our animal findings using the embryonically neural crest-originated PC12 cell model. We found that stimulation of the IgG immune complex led to increased levels of FcγRI and inflammatory mediators, which were attenuated by the anti-FcγRI antibody in these cells. Collectively, our results from animal and cell-based studies suggest that FcγRI is a critical player for peripheral nerve injury-induced neuropathic pain by mediating pain-related immunological events, which therefore may provide a new therapeutic target for protection against chronic pain.
A pilot study of alternative TrkAIII splicing in Merkel cell carcinoma: a potential oncogenic mechanism and novel therapeutic target
Authors: Cappabianca Lucia , Guadagni Stefano , Maccarone Rita , Sebastiano Michela , Chiominto Alessandro , Farina Antonietta Rosella , Mackay Andrew Reay
Published: October 2019

Background

Merkel cell carcinomas (MCCs) are rare, aggressive, therapeutically-challenging skin tumours that are increasing in incidence and have poor survival rates. The majority are caused by genomic Merkel cell polyomavirus (MCPyV) integration and MCPyV T-antigen expression. Recently, a potential oncogenic role for the tropomyosin-related tyrosine kinase A receptor (TrkA) has been proposed in MCC. Alternative TrkAIII splicing is a TrkA oncogenic activation mechanism that can be promoted by SV40 large T-antigen, an analogue of MCPyV large T-antigen. In this pilot study, therefore, we have evaluated TrkAIII splicing as a novel potential oncogenic mechanism and therapeutic target in MCPyV positive MCC.

Methods

Formalin-fixed paraffin-embedded MCC tissues, consisting of 10 stage IV, 1 stage IIIB, 1 stage IIB, 4 stage IIA and 2 stage I tumours, from patients diagnosed and treated from September 2006 to March, 2019, at the University of L’Aquila, L’Aquila, Italy, were compared to 3 primary basal cell carcinomas (BCCs), 3 primary squamous cell carcinomas (SCCs) and 2 normal skin samples by RT-PCR for MCPyV large T-antigen, small T-antigen, VP-1 expression and alternative TrkAIII splicing and by indirect IF for evidence of intracellular TrkA isoform expression and activation.

Results

9 of 10 Recurrent stage IV MCCs were from patients (P.1–3) treated with surgery plus loco-regional Melphalan chemotherapy and remaining MMCs, including 1 stage IV tumour, were from patients treated with surgery alone (P. 4–11). All MCPyV positive MCCs exhibiting MCPyV large T-antigen expression (17 of 18MCCs, 90%) exhibited alternative TrkAIII mRNA splicing (100%), which was exclusive in a significant number and predominant (> 50%) in all stage IV MCCs and the majority of stage 1-III MCCs. MCCs with higher TrkAIII to 18S rRNA expression ratios also exhibited strong or intermediate immunoreactivity to anti-TrkA antibodies, consistent with cytoplasmic TrkAIII expression and activation. In contrast, the MCPyV negative MCC, BCCs, SCCs and normal skin tissues all exhibited exclusive fully-spliced TrkA mRNA expression, associated with variable immunoreactivity for non-phosphorylated but not phosphorylated TrkA.

Conclusions

MCPyV positive MCCs but not MCPyV negative MCC, BCCs and SCCs exhibit predominant alternative TrkAIII splicing, with evidence of intracellular TrkAIII activation. This establishes a new potential MCC subset, unveils a novel potential MCPyV oncogenic mechanism and identifies TrkAIII as a novel potential therapeutic target in MCPyV positive MCC.
Prevalence and spectrum of MLH1, MSH2, and MSH6 pathogenic germline variants in Pakistani colorectal cancer patients
Authors: Rashid Muhammad Usman , Naeemi Humaira , Muhammad Noor , Loya Asif , Lubiński Jan , Jakubowska Anna , Yusuf Muhammed Aasim
Published: October 2019

Background

Pathogenic germline variants in MLH1, MSH2 and MSH6 genes account for the majority of Lynch syndrome (LS). In this first report from Pakistan, we investigated the prevalence of pathogenic MLH1/MSH2/MSH6 variants in colorectal cancer (CRC) patients.

Methods

Consecutive cases (n = 212) were recruited at the Shaukat Khanum Memorial Cancer Hospital and Research Centre (SKMCH&RC), between November 2007 to March 2011. Patients with a family history of > 3 or 2 HNPCC-associated cancers were classified as HNPCC (n = 9) or suspected-HNPCC (n = 20), respectively (group 1; n = 29). Cases with no family history were designated as non-HNPCC (group 2; n = 183). MLH1/MSH2/MSH6 genes were comprehensively screened in group 1. Pathogenic/likely pathogenic variants identified in group 1 were subsequently evaluated in group 2.

Results

Eight distinct pathogenic/likely pathogenic MLH1/MSH2 variants were found in group 1 (10/29; 34.5%), belonging to HNPCC (5/9; 55.6%) and suspected-HNPCC (5/20; 25%) families and in group 2 (2/183; 1.1%) belonging to non-HNPCC. Overall, three recurrent variants (MSH2 c.943-1G > C, MLH1 c.1358dup and c.2041G > A) accounted for 58.3% (7/12) of all families harboring pathogenic/likely pathogenic MLH1/MSH2 variants. Pathogenic MSH6 variants were not detected.

Conclusion

Pathogenic/likely pathogenic MLH1/MSH2 variants account for a substantial proportion of CRC patients with HNPCC/suspected-HNPCC in Pakistan. Our findings suggest that HNPCC/suspected-HNPCC families should be tested for these recurrent variants prior to comprehensive gene screening in this population.
Enhanced recovery after surgery (ERAS) pathway for primary hip and knee arthroplasty: study protocol for a randomized controlled trial
Authors: Li Jingyi , Zhu Haibei , Liao Ren
Published: October 2019

Background

With the substantially growing trend of the aging populations in China and the rest of the world, the number of total hip and total knee arthroplasty (THA and TKA) cases are increasing dramatically. It is important to develop practical strategies to improve the quality of healthcare and better outcome for patients undergoing THA and TKA. Enhanced recovery after surgery (ERAS) pathways have been reported to promote earlier recovery and be beneficial for patients. We propose the hypothesis that the ERAS pathway could reduce the length of stay (LOS) in hospital for patients undergoing primary THA or TKA.

Methods/Design

This trial is a prospective, open-labelled, multi-centered, randomized controlled trial that will test the superiority of the ERAS pathway in term of LOS in hospital for the patients undergoing primary THA or TKA compared to current non-ERAS clinical practice. A total of 640 patients undergoing primary THA or TKA will be randomly allocated to either ERAS pathway (ERAS group) or conventional care according to individual participating center (non-ERAS group). The primary outcome is the total LOS in hospital; the secondary outcomes include postoperative LOS, all-cause mortality by 30 days after operation, in-hospital complications, early mobilization, postoperative pain control, total in-hospital cost, and readmission rate by 30 days after discharge from the hospital.

Discussion

This trial is designed to evaluate the superiority of the ERAS pathway to conventional non-ERAS clinical practice in reducing the LOS. The results may provide new insight into the clinical applications of the ERAS pathway for THA and TKA.

Trial registration

National Institutes of Health Clinical Trials Registry, NCT03517098. Registered on 4 May 2018.
Molecular investigation and phylogeny of species of the Anaplasmataceae infecting animals and ticks in Senegal
Authors: Dahmani Mustapha , Davoust Bernard , Sambou Masse , Bassene Hubert , Scandola Pierre , Ameur Tinhinene , Raoult Didier , Fenollar Florence , Mediannikov Oleg
Published: October 2019

Background

Our study aimed to assess the diversity of the species of Anaplasmataceae in Senegal that infect animals and ticks in three areas: near Keur Momar Sarr (northern region), Dielmo and Diop (Sine Saloum, central region of Senegal), and in Casamance (southern region of Senegal).

Methods

A total of 204 ticks and 433 blood samples were collected from ruminants, horses, donkeys and dogs. Ticks were identified morphologically and by molecular characterization targeting the 12S rRNA gene. Molecular characterization of species of Anaplasmataceae infecting Senegalese ticks and animals was conducted using the 23S rRNA, 16S rRNA, rpoB and groEL genes.

Results

Ticks were identified as Rhipicephalus evertsi evertsi (84.3%), Hyalomma rufipes (8.3%), Hyalomma impeltatum (4.9%), R. bursa (1.5%) and R. muhsamae (0.9%). The overall prevalence of Anaplasmataceae infection in ticks was 0.9%, whereas 41.1% of the sampled animals were found infected by one of the species belonging to this family. We identified the pathogen Anaplasma ovis in 55.9% of sheep, A. marginale and A. centrale in 19.4% and 8.1%, respectively, of cattle, as well as a putative new species of Anaplasmataceae. Two Anaplasma species commonly infecting ruminants were identified. Anaplasma cf. platys, closely related to A. platys was identified in 19.8% of sheep, 27.7% of goats and 22.6% of cattle, whereas a putative new species, named here provisionally “Candidatus Anaplasma africae”, was identified in 3.7% of sheep, 10.3% of goats and 8.1% of cattle. Ehrlichia canis and Anaplasma platys were identified only from dogs sampled in the Keur Momar Sarr area. Ehrlichia canis was identified in 18.8% of dogs and two R. e. evertsi ticks removed from the same sheep. Anaplasma platys was identified in 15.6% of dogs. Neither of the dogs sampled from Casamance region nor the horses and donkeys sampled from Keur Momar Sarr area were found infected by an Anaplasmataceae species.

Conclusions

This study presents a summary of Anaplasmataceae species that infect animals and ticks in three areas from the northern, central and southern regions of Senegal. To our knowledge, our findings demonstrate for the first time the presence of multiple Anaplasmataceae species that infect ticks and domestic animals in Senegal. We recorded two potentially new species commonly infecting ruminants named here provisionally as Anaplasma cf. platys and “Candidatus Anaplasma africae”. However, E. canis was the only species identified and amplified from ticks. None of the other Anaplasmataceae species identified in animals were identified in the tick species collected from animals.