bluuebunny/crossref_metadata_2025 huggingface dataset:
Four mutations in MITF, SOX10 and PAX3 genes were identified as genetic causes of waardenburg syndrome in four unrelated Iranian patients: case report
Authors: Zardadi Safoura , Rayat Sima , Doabsari Maryam Hassani , Alishiri Aliagha , Keramatipour Mohammad , Shahri Zeynab Javanfekr , Morovvati Saeid
Published: February 2021

Background

Waardenburg syndrome (WS) is a rare genetic disorder. The purpose of this study was to investigate clinical and molecular characteristics of WS in four probands from four different Iranian families.

Case presentation

The first patient was a 1-year-old symptomatic boy with congenital hearing loss and heterochromia iridis with a blue segment in his left iris. The second case was a 1.5-year-old symptomatic girl who manifested congenital profound hearing loss, brilliant blue eyes, and skin hypopigmentation on the abdominal region at birth time. The third patient was an 8-month-old symptomatic boy with developmental delay, mild atrophy, hypotonia, brilliant blue eyes, skin hypopigmentation on her hand and foot, Hirschsprung disease, and congenital profound hearing loss; the fourth patient was a 4-year-old symptomatic boy who showed dystopia canthorum, broad nasal root, synophrys, skin hypopigmentation on her hand and abdomen, brilliant blue eyes, and congenital profound hearing loss. Whole exome sequencing (WES) was used for each proband to identify the underlying genetic factor. Sanger sequencing was performed for validation of the identified mutations in probands and the available family members. A novel heterozygous frameshift mutation, c.996delT (p.K334Sfs*15), on exon 8 of the MITF gene was identified in the patient of the first family diagnosed with WS2A. Two novel de novo heterozygous mutations including a missense mutation, c.950G > A (p.R317K), on exon 8 of the MITF gene, and a frameshift mutation, c.684delC (p.E229Sfs*57), on the exon 3 of the SOX10 gene were detected in patients of the second and third families with WS2A and PCWH (Peripheral demyelinating neuropathy, Central dysmyelinating leukodystrophy, Waardenburg syndrome, Hirschsprung disease), respectively. A previously reported heterozygous frameshift mutation, c.1024_1040del AGCACGATTCCTTCCAA, (p.S342Pfs*62), on exon 7 of the PAX3 gene was identified in the patient of the fourth family with WS1.

Conclusions

An exact description of the mutations responsible for WS provides useful information to explain the molecular cause of clinical features of WS and contributes to better genetic counseling of WS patients and their families.
Posttraumatic stress disorder among adolescents in Brazil: a cross-sectional study
Authors: Avanci Joviana Quintes , Serpeloni Fernanda , de Oliveira Thiago Pires , de Assis Simone Gonçalves
Published: February 2021

Background

The frequency of trauma and different types of violence exposure in urban areas and their effects on the mental health of adolescents in developing countries are poorly investigated. Most information about traumatized young people comes from war scenarios or disasters. This study aimed to determine the prevalence of PTSD in trauma-exposed students in a low-resource city of the state of Rio de Janeiro, Brazil. The effects of sociodemographic and individual and family factors in the development of PTSD were also investigated.

Methods

Through multi-stage cluster sampling, 862 adolescents (Mage = 15 years old, 65% female) from public and private schools in the city of São Gonçalo were selected for the study. Self-rating structured questionnaires were applied to assess sociodemographic profile, exposure to physical and psychological violence (family, school, community), sexual abuse, social support, social functional impairment, resilience, and posttraumatic stress disorder. The data were grouped in blocks regarding sociodemographic, individual, family, and community variables. For statistical analysis, chi-square, Fisher’s exact test, and logistic regression were performed.

Results

The PTSD prevalence was 7.8% among adolescents. Boys were exposed to significantly higher number of events of community violence, while girls to family violence. The adjusted odds ratio (OR) for PTSD were statistically significant for age (OR, 1.45, [95% CI, 1.043–2.007]), social functional impairment (OR, 4.82, [95% CI, 1.77–13.10]), severe maternal physical violence (OR, 2.79, [95% CI, 0.79–9.93]), psychological violence by significant people (OR, 3.96, [95% CI, 1.89–8.31]) and a high number of episodes of community violence (OR, 3.52, [95% CI, 1.47–8.40).

Conclusions

There was a high prevalence of PTSD within this population associated with exposure to violence. Not only physical, but also psychological violence contributed to PTSD. The results also raise awareness to the differences in life trajectories between boys and girls regarding violence. These differences need to be better understood in order to enable the development of effective preventative interventions. Treating and preventing mental health disorders presents a challenge for countries, especially those with a lower degree of social and economic development and high community violence.
Dysplasia epiphysealis hemimelica of the radial head: a rare case report
Authors: Mohamadian Hiva , Bahardoust Mansour , Sanandaji Borzouyeh Molazem , Saberi Saba
Published: February 2021

Background

Dysplasia epiphysealis hemimelica (DEH) is a rare benign overgrowth generally affecting the epiphyses and short bones of the lower limbs. DEH in the elbow joint is extremely rare, and to date, only three cases of DEH have been reported in the radial head.

Case presentation

In this study, we report a case of DEH located in the radial head of the right elbow of a 10-year-old boy, which was presented with elbow pain and limited range of motion. In clinical examination, an asymmetrical enlargement was observed over the elbow. The lesion was resected surgically, and the patient’s symptoms resolved afterward. The histologic analysis of the lesion confirmed the diagnosis of DEH.

Conclusion

This report highlights the role of DEH in the differential diagnosis of elbow pathologies, particularly its differentiation from osteochondroma.
Utilising the perspectives of patients with lower-limb osteoarthritis on prescribed physical activity to develop a theoretically informed physiotherapy intervention
Authors: Willett Matthew , Greig Carolyn , Fenton Sally , Rogers David , Duda Joan , Rushton Alison
Published: February 2021

Background

Lower-limb osteoarthritis (OA) causes high levels of pain and disability. Physiotherapists are the primary healthcare provider of non-pharmacological treatments, and incorporate strategies to optimise physical activity (PA) to aid patients with lower-limb OA to moderate their clinical symptoms. However, patients with lower-limb OA have low adherence to PA recommendations both during treatment and after discharge. This study aimed to use knowledge of identified barriers and facilitators to physiotherapy prescribed PA (during treatment and post-discharge) to develop a theoretically informed intervention to optimise adherence to PA for patients with lower-limb OA during treatment and post-discharge.

Methods

1) A purposive sample of 13 patients with lower-limb OA participated in semi-structured interviews following physiotherapy treatment. Inductive analysis identified themes/subthemes reflecting barriers and facilitators to physiotherapist prescribed PA, which were organised deductively according to personal factors, treatment and post-discharge phases.

2) Themes/subthemes were mapped onto the theoretical domains framework (TDF).

3) Behaviour change techniques (BCTs) were coded from the key identified domains and a theoretically informed physiotherapy intervention addressing barriers and using facilitators, was developed.

Results

Themes of patient confidence, mind-set, motivation, OA symptoms and PA experiences were primary personal factors that influenced PA adherence; with the TDF domain ‘Beliefs about capabilities’ most important to target. During treatment, the theme of routine formation was the major driver of personal factors; and primarily influenced by developing a positive physiotherapist-patient relationship. Post-discharge, physical factors, psychosocial factors and ongoing access to resources were important themes influencing PA maintenance. ‘Environmental context and resources’ and ‘social influences’ emerged as the key TDF domains to target during treatment and post-discharge. The proposed theoretically informed intervention included 26 BCTs delivered across conceptual phases of adoption, routine formation, and maintenance.

Conclusion

A theoretically informed physiotherapy intervention was proposed to optimise PA adherence in patients with lower-limb OA. The included BCTs primarily target patients’ perceived beliefs about their capabilities, by developing a PA routine during treatment and facilitating appropriate psychosocial support and access to resources for PA maintenance post-discharge. The feasibility of delivering the intervention in clinical practice will now be evaluated.
Identification of S1PR3 gene signature involved in survival of sepsis patients
Authors: Feng Anlin , Ma Wenli , Faraj Reem , Kelly Gabriel T. , Black Stephen M. , Fallon Michael B. , Wang Ting
Published: February 2021

Background

Sepsis is a life-threatening complication of infection that rapidly triggers tissue damage in multiple organ systems and leads to multi-organ deterioration. Up to date, prognostic biomarkers still have limitations in predicting the survival of patients with sepsis. We need to discover more prognostic biomarkers to improve the sensitivity and specificity of the prognosis of sepsis patients. Sphingosine-1-phosphate (S1P) receptor 3 (S1PR3), as one of the S1P receptors, is a prospective prognostic biomarker regulating sepsis-relevant events, including compromised vascular integrity, antigen presentation, and cytokine secretion. Until now, no S1PR3-related prognostic gene signatures for sepsis patients have been found.

Methods

This study intends to obtain an S1PR3-associated gene signature from whole blood samples to be utilized as a probable prognostic tool for patients with sepsis.

Results

We obtained an 18-gene S1PR3-related molecular signature (S3MS) from the intersection of S1PR3-associated genes and survival-associated genes. Numerous important immunity pathways that regulate the progression of sepsis are enriched among our 18 genes. Significantly, S3MS functions greatly in both the discovery and validation cohort. Furthermore, we demonstrated that S3MS obtains significantly better classification performance than random 18-gene signatures.

Conclusions

Our results confirm the key role of S1PR3-associated genes in the development of sepsis, which will be a potential prognostic biomarker for patients with sepsis. Our results also focus on the classification performance of our S3MS as biomarkers for sepsis, which could also provide an early warning system for patients with sepsis.
Rapid ethnographic assessment for potential anti-malarial mass drug administration in an outbreak area of Santo Domingo, Dominican Republic
Authors: Keys Hunter , Ureña Keyla , Reyes Jhefres , Bardosh Kevin , Pell Christopher , Puello Jose , Blount Stephen , Noland Gregory S.
Published: February 2021

Background

In the Dominican Republic, a recent outbreak of malaria in the capital, Santo Domingo, threatens efforts to eliminate the disease. Mass drug administration (MDA) has been proposed as one strategy to reduce transmission. The success of MDA is contingent upon high levels of acceptance among the target population. To inform the design of future MDA campaigns, this rapid ethnographic assessment examined malaria-related knowledge and attitudes toward malaria MDA among residents of a transmission focus in Santo Domingo.

Methods

In October 2019, a rapid ethnographic assessment was conducted in the Los Tres Brazos transmission focus, which had not previously received MDA. National malaria programme staff conducted 61 structured interviews with key informants, recorded observations, and held 72 informal conversations. Using a grounded theory approach, data were analysed during three workshop sessions with research team members.

Results

Among those who had heard of malaria in the structured interviews (n = 39/61; 64%), understanding of the disease was largely based on personal experience from past outbreaks or through word-of-mouth. Community health workers (promotores) were trusted for health information and malaria diagnosis more so than professional clinicians. No participant (0%) was familiar with malaria MDA. After learning about MDA, almost all study participants (92%) said that they would participate, seeing it as a way to care for their community. Reasons for not participating in future MDA included not trusting drug administrators, feeling reluctant to take unprescribed medicine, and fear of missing work. Additional identified challenges to MDA included reaching specific demographic groups, disseminating effective MDA campaign messages, and managing misinformation and political influence.

Conclusion

Residents appear accepting of MDA despite a lack of prior familiarity. Successful MDA will depend on several factors: fostering relationships among community-based health workers, clinicians, community leaders, and others; developing clear health messages that use local terms and spreading them through a variety of media and social networks; and contextualizing MDA as part of a broader effort to promote community health.
Low prevalence of highly sulfadoxine‐resistant dihydropteroate synthase alleles in Plasmodium falciparum isolates in Benin
Authors: Svigel Samaly Souza , Adeothy Adicath , Kpemasse Augustin , Houngbo Ernest , Sianou Antoine , Saliou Ramani , Patton Monica E. , Dagnon Fortune , Halsey Eric S. , Tchevoede Alexis , Udhayakumar Venkatachalam , Lucchi Naomi W.
Published: February 2021

Background

In 2004, in response to high levels of treatment failure associated with sulfadoxine-pyrimethamine (SP) resistance, Benin changed its first-line malaria treatment from SP to artemisinin-based combination therapy for treatment of uncomplicated Plasmodium falciparum malaria. Resistance to SP is conferred by accumulation of single nucleotide polymorphisms (SNPs) in P. falciparum genes involved in folate metabolism, dihydrofolate reductase (Pfdhfr) and dihydropteroate synthase (Pfdhps), targeted by pyrimethamine and sulfadoxine, respectively. Because SP is still used for intermittent preventive treatment in pregnant women (IPTp) and seasonal malaria chemoprevention (SMCP) in Benin, the prevalence of Pfdhfr and Pfdhps SNPs in P. falciparum isolates collected in 2017 were investigated.

Methods

This study was carried out in two sites where the transmission of P. falciparum malaria is hyper-endemic: Klouékanmey and Djougou. Blood samples were collected from 178 febrile children 6–59 months old with confirmed uncomplicated P. falciparum malaria and were genotyped for SNPs associated with SP resistance.

Results

The Pfdhfr triple mutant IRN (N51I, C59R, and S108N) was the most prevalent (84.6%) haplotype and was commonly found with the Pfdhps single mutant A437G (50.5%) or with the Pfdhps double mutant S436A and A437G (33.7%). The quintuple mutant, PfdhfrIRN/PfdhpsGE (A437G and K540E), was rarely observed (0.8%). The A581G and A613S mutant alleles were found in 2.6 and 3.9% of isolates, respectively. Six isolates (3.9%) were shown to harbour a mutation at codon I431V, recently identified in West African parasites.

Conclusions

This study showed that Pfdhfr triple IRN mutants are near fixation in this population and that the highly sulfadoxine-resistant Pfdhps alleles are not widespread in Benin. These data support the continued use of SP for chemoprevention in these study sites, which should be complemented by periodic nationwide molecular surveillance to detect emergence of resistant genotypes.
Four decades of measuring stillbirths and neonatal deaths in Demographic and Health Surveys: historical review
Authors: Akuze Joseph , Cousens Simon , Lawn Joy E. , Waiswa Peter , Gordeev Vladimir Sergeevich , Arnold Fred , Croft Trevor , Baschieri Angela , Blencowe Hannah
Published: February 2021

Background

Worldwide, an estimated 5.1 million stillbirths and neonatal deaths occur annually, 98% in low- and middle-income countries. Limited coverage of civil and vital registration systems necessitates reliance on women’s retrospective reporting in household surveys for data on these deaths. The predominant platform, Demographic and Health Surveys (DHS), has evolved over the last 35 years and differs by country, yet no previous study has described these differences and the effects of these changes on stillbirth and neonatal death measurement.

Methods

We undertook a review of DHS model questionnaires, protocols and methodological reports from DHS-I to DHS-VII, focusing on the collection of information on stillbirth and neonatal deaths describing differences in approaches, questionnaires and geographic reach up to December 9, 2019. We analysed the resultant data, applied previously used data quality criteria including ratios of stillbirth rate (SBR) to neonatal mortality rate (NMR) and early NMR (ENMR) to NMR, comparing by country, over time and by DHS module.

Results

DHS has conducted >320 surveys in 90 countries since 1984. Two types of maternity history have been used: full birth history (FBH) and full pregnancy history (FPH). A FBH collecting information only on live births has been included in all model questionnaires to date, with data on stillbirths collected through a reproductive calendar (DHS II-VI) or using additional questions on non-live births (DHS-VII). FPH collecting information on all pregnancies including live births, miscarriages, abortions and stillbirths has been used in 17 countries. We found no evidence of variation in stillbirth data quality assessed by SBR:NMR over time for FBH surveys with reproductive calendar, some variation for surveys with FBH in DHS-VII and most variation among the surveys conducted with a FPH. ENMR:NMR ratio increased over time, which may reflect changes in data quality or real epidemiological change.

Conclusion

DHS remains the major data source for pregnancy outcomes worldwide. Although the DHS model questionnaire has evolved over the last three and half decades, more robust evidence is required concerning optimal methods to obtain accurate data on stillbirths and neonatal deaths through household surveys and also to develop and test standardised data quality criteria.
Termination of pregnancy data completeness and feasibility in population-based surveys: EN-INDEPTH study
Authors:   the Every Newborn-INDEPTH Study Collaborative Group, Enuameh Yeetey Akpe Kwesi , Dzabeng Francis , Blencowe Hannah , Thysen Sanne M. , Abebe Solomon Mekonnen , Asante Kwaku Poku , Tawiah Charlotte , Gordeev Vladimir Sergeevich , Adeapena Wisdom , Kwesiga Doris , Kasasa Simon , Zandoh Charles , Imam Md. Ali , Amenga-Etego Seeba , Newton Sam K. , Owusu-Agyei Seth , Lawn Joy E. , Waiswa Peter , Cresswell Jenny A.
Published: February 2021

Background

Termination of pregnancy (TOP) is a common cause of maternal morbidity and mortality in low- and middle-income countries. Population-based surveys are the major data source for TOP data in LMICs but are known to have shortcomings that require improving. The EN-INDEPTH multi-country survey employed a full pregnancy history approach with roster and new questions on TOP and Menstrual Restoration. This mixed methods paper assesses the completeness of responses to questions eliciting TOP information from respondents and reports on practices, barriers, and facilitators to TOP reporting.

Methods

The EN-INDEPTH study was a population-based cross-sectional study. The Full Pregnancy History arm of the study surveyed 34,371 women of reproductive age between 2017 and 2018 in five Health and Demographic Surveillance System (HDSS) sites of the INDEPTH network: Bandim, Guinea-Bissau; Dabat, Ethiopia; IgangaMayuge, Uganda; Kintampo, Ghana; and Matlab, Bangladesh. Completeness and time spent in answering TOP questions were evaluated using simple tabulations and summary statistics. Exact binomial 95% confidence intervals were computed for TOP rates and ratios. Twenty-eight (28) focus group discussions were undertaken and analysed thematically.

Results

Completeness of responses regarding TOP was between 90.3 and 100.0% for all question types. The new questions elicited between 2.0% (1.0–3.4), 15.5% (13.9–17.3), and 11.5% (8.8–14.7) lifetime TOP cases over the roster questions from Dabat, Ethiopia; Matlab, Bangladesh; and Kintampo, Ghana, respectively. The median response time on the roster TOP questions was below 1.3 minutes in all sites. Qualitative results revealed that TOP was frequently stigmatised and perceived as immoral, inhumane, and shameful. Hence, it was kept secret rendering it difficult and uncomfortable to report. Miscarriages were perceived to be natural, being easier to report than TOP. Interviewer techniques, which were perceived to facilitate TOP disclosure, included cultural competence, knowledge of contextually appropriate terms for TOP, adaptation to interviewee’s individual circumstances, being non-judgmental, speaking a common language, and providing detailed informed consent.

Conclusions

Survey roster questions may under-represent true TOP rates, since the new questions elicited responses from women who had not disclosed TOP in the roster questions. Further research is recommended particularly into standardised training and approaches to improving interview context and techniques to facilitate TOP reporting in surveys.
Stillbirth outcome capture and classification in population-based surveys: EN-INDEPTH study
Authors:   the Every Newborn-INDEPTH Study Collaborative Group, Blencowe Hannah , Bottecchia Matteo , Kwesiga Doris , Akuze Joseph , Haider M. Moinuddin , Galiwango Edward , Dzabeng Francis , Fisker Ane B. , Enuameh Yeetey Akpe Kwesi , Geremew Bisrat Misganaw , Nareeba Tryphena , Woodd Susannah , Beedle Alexandra , Peven Kimberly , Cousens Simon , Waiswa Peter , Lawn Joy E.
Published: February 2021

Background

Household surveys remain important sources of stillbirth data, but omission and misclassification are common. Classifying adverse pregnancy outcomes as stillbirths requires accurate reporting of vital status at birth and gestational age or birthweight for every pregnancy. Further categorisation, e.g. by sex, or timing (intrapartum/antepartum) improves data to understand and prevent stillbirth.

Methods

We undertook a cross-sectional population-based survey of women of reproductive age in five health and demographic surveillance system sites in Bangladesh, Ethiopia, Ghana, Guinea-Bissau and Uganda (2017–2018). All women answered a full birth history with pregnancy loss questions (FBH+) or a full pregnancy history (FPH). A sub-sample across both groups were asked additional stillbirth questions. Questions were evaluated using descriptive measures. Using an interpretative paradigm and phenomenology methodology, focus group discussions with women exploring barriers to reporting birthweight for stillbirths were conducted. Thematic analysis was guided by an a priori codebook.

Results

Overall 69,176 women reported 98,483 livebirths (FBH+) and 102,873 pregnancies (FPH). Additional questions were asked for 1453 stillbirths, 1528 neonatal deaths and 12,620 surviving children born in the 5 years prior to the survey. Completeness was high (> 99%) for existing FBH+/FPH questions on signs of life at birth and gestational age (months). Discordant responses in signs of life at birth between different questions were common; nearly one-quarter classified as stillbirths on FBH+/FPH were reported born alive on additional questions. Availability of information on gestational age (weeks) (58.1%) and birthweight (13.2%) was low amongst stillbirths, and heaping was common. Most women (93.9%) were able to report the sex of their stillborn baby. Response completeness for stillbirth timing (18.3–95.1%) and estimated proportion intrapartum (15.6–90.0%) varied by question and site. Congenital malformations were reported in 3.1% stillbirths. Perceived value in weighing a stillborn baby varied and barriers to weighing at birth a nd knowing birthweight were common.

Conclusions

Improving stillbirth data in surveys will require investment in improving the measurement of vital status, gestational age and birthweight by healthcare providers, communication of these with women, and overcoming reporting barriers. Given the large burden and effect on families, improved data must be made available to end preventable stillbirths.
Stillbirth maternity care measurement and associated factors in population-based surveys: EN-INDEPTH study
Authors:   and the Every Newborn-INDEPTH Study Collaborative Group, Di Stefano Lydia , Bottecchia Matteo , Yargawa Judith , Akuze Joseph , Haider M. Moinuddin , Galiwango Edward , Dzabeng Francis , Fisker Ane B. , Geremew Bisrat Misganaw , Cousens Simon , Lawn Joy E. , Blencowe Hannah , Waiswa Peter
Published: February 2021

Background

Household surveys remain important sources of maternal and child health data, but until now, standard surveys such as Demographic and Health Surveys (DHS) have not collected information on maternity care for women who have experienced a stillbirth. Thus, nationally representative data are lacking to inform programmes to address the millions of stillbirths which occur annually.

Methods

The EN-INDEPTH population-based survey of women of reproductive age was undertaken in five Health and Demographic Surveillance System sites in Bangladesh, Ethiopia, Ghana, Guinea-Bissau and Uganda (2017–2018). All women answered a full birth history with additional questions on pregnancy losses (FBH+) or full pregnancy history (FPH). A sub-sample, including all women reporting a recent stillbirth or neonatal death, was asked additional maternity care questions. These were evaluated using descriptive measures. Associations between stillbirth and maternal socio-demographic characteristics, babies’ characteristics and maternity care use were assessed using a weighted logistic regression model for women in the FBH+ group.

Results

A total of 15,591 women reporting a birth since 1 January 2012 answered maternity care questions. Completeness was very high (> 99%), with similar proportions of responses for both live and stillbirths. Amongst the 14,991 births in the FBH+ group, poorer wealth status, higher parity, large perceived baby size-at-birth, preterm or post-term birth, birth in a government hospital compared to other locations and vaginal birth were associated with increased risk of stillbirth after adjusting for potential confounding factors. Regarding association with reported postnatal care, women with a stillbirth were more likely to report hospital stays of > 1 day. However, women with a stillbirth were less likely to report having received a postnatal check compared to those with a live birth.

Conclusions

Women who had experienced stillbirth were able to respond to questions about pregnancy and birth, and we found no reason to omit questions to these women in household surveys. Our analysis identified several potentially modifiable factors associated with stillbirth, adding to the evidence-base for policy and action in low- and middle-income contexts. Including these questions in DHS-8 would lead to increased availability of population-level data to inform action to end preventable stillbirths.
Efficient CRISPR/Cas9 mediated Pooled-sgRNAs assembly accelerates targeting multiple genes related to male sterility in cotton
Authors: Ramadan Mohamed , Alariqi Muna , Ma Yizan , Li Yanlong , Liu Zhenping , Zhang Rui , Jin Shuangxia , Min Ling , Zhang Xianlong
Published: February 2021

Background

Upland cotton (Gossypium hirsutum), harboring a complex allotetraploid genome, consists of A and D sub-genomes. Every gene has multiple copies with high sequence similarity that makes genetic, genomic and functional analyses extremely challenging. The recent accessibility of CRISPR/Cas9 tool provides the ability to modify targeted locus efficiently in various complicated plant genomes. However, current cotton transformation method targeting one gene requires a complicated, long and laborious regeneration process. Hence, optimizing strategy that targeting multiple genes is of great value in cotton functional genomics and genetic engineering.

Results

To target multiple genes in a single experiment, 112 plant development-related genes were knocked out via optimized CRISPR/Cas9 system. We optimized the key steps of pooled sgRNAs assembly method by which 116 sgRNAs pooled together into 4 groups (each group consisted of 29 sgRNAs). Each group of sgRNAs was compiled in one PCR reaction which subsequently went through one round of vector construction, transformation, sgRNAs identification and also one round of genetic transformation. Through the genetic transformation mediated Agrobacterium, we successfully generated more than 800 plants. For mutants identification, Next Generation Sequencing technology has been used and results showed that all generated plants were positive and all targeted genes were covered. Interestingly, among all the transgenic plants, 85% harbored a single sgRNA insertion, 9% two insertions, 3% three different sgRNAs insertions, 2.5% mutated sgRNAs. These plants with different targeted sgRNAs exhibited numerous combinations of phenotypes in plant flowering tissues.

Conclusion

All targeted genes were successfully edited with high specificity. Our pooled sgRNAs assembly offers a simple, fast and efficient method/strategy to target multiple genes in one time and surely accelerated the study of genes function in cotton.
Rapid tannin profiling of tree fodders using untargeted mid-infrared spectroscopy and partial least squares regression
Authors: Ortuño Jordi , Stergiadis Sokratis , Koidis Anastasios , Smith Jo , Humphrey Chris , Whistance Lindsay , Theodoridou Katerina
Published: February 2021

Background

The presence of condensed tannins (CT) in tree fodders entails a series of productive, health and ecological benefits for ruminant nutrition. Current wet analytical methods employed for full CT characterisation are time and resource-consuming, thus limiting its applicability for silvopastoral systems. The development of quick, safe and robust analytical techniques to monitor CT’s full profile is crucial to suitably understand CT variability and biological activity, which would help to develop efficient evidence-based decision-making to maximise CT-derived benefits. The present study investigates the suitability of Fourier-transformed mid-infrared spectroscopy (MIR: 4000–550 cm−1) combined with multivariate analysis to determine CT concentration and structure (mean degree of polymerization—mDP, procyanidins:prodelphidins ratio—PC:PD and cis:trans ratio) in oak, field maple and goat willow foliage, using HCl:Butanol:Acetone:Iron (HBAI) and thiolysis-HPLC as reference methods.

Results

The MIR spectra obtained were explored firstly using Principal Component Analysis, whereas multivariate calibration models were developed based on partial least-squares regression. MIR showed an excellent prediction capacity for the determination of PC:PD [coefficient of determination for prediction (R2P) = 0.96; ratio of prediction to deviation (RPD) = 5.26, range error ratio (RER) = 14.1] and cis:trans ratio (R2P = 0.95; RPD = 4.24; RER = 13.3); modest for CT quantification (HBAI: R2P = 0.92; RPD = 3.71; RER = 13.1; Thiolysis: R2P = 0.88; RPD = 2.80; RER = 11.5); and weak for mDP (R2P = 0.66; RPD = 1.86; RER = 7.16).

Conclusions

MIR combined with chemometrics allowed to characterize the full CT profile of tree foliage rapidly, which would help to assess better plant ecology variability and to improve the nutritional management of ruminant livestock.
Single institutional outcomes of whole brain radiotherapy for metastatic melanoma brain metastases
Authors: Jiang Cecilia , Kleber Troy J. , Switchenko Jeffrey M. , Khan Mohammad K.
Published: February 2021

Background

The management of melanoma with brain metastases (MBM) is increasingly complex, especially given recent improvements in targeted agents, immunotherapy, and radiotherapy. Whole brain radiation therapy (WBRT) is a longstanding radiotherapy technique for which reported patient outcomes and experiences are limited. We sought to report our institutional outcomes for MBM patients receiving WBRT and assess whether other clinical factors impact prognosis.

Methods

A retrospective review of a single institution database was performed. Patients diagnosed with MBM from 2000 to 2018 treated with WBRT, with or without other systemic treatments, were included. Post-WBRT brain MRI scans were assessed at timed intervals for radiographic response. Clinical and treatment variables associated with overall survival (OS), distant failure-free survival (DFFS), local failure-free survival (LFFS), and progression-free survival (PFS) were assessed. Data on radiation-induced side effects, including radionecrosis, hemorrhage, and memory deficits, was also captured.

Results

63 patients with MBM were ultimately included in our study. 69% of patients had 5 or more brain metastases at the time of WBRT, and 68% had extracranial disease. The median dose of WBRT was 30 Gy over 10 fractions. Median follow-up was 4.0 months. Patients receiving WBRT had a median OS of 7.0 months, median PFS of 2.2 months, median DFFS of 6.1 months, and median LFFS of 4.9 months. Performance status correlated with OS on both univariate and multivariable analysis. BRAF inhibitor was the only systemic therapy to significantly impact OS on univariate analysis (HR 0.24, 95% CI 0.07–0.79, p = 0.019), and this effect extended to multivariable analysis as well. Post-WBRT intralesional hemorrhage decreased DFFS on both univariate and multivariable analysis. Of patients with post-treatment brain scans available, there was a 16% rate of radionecrosis, 32% rate of hemorrhage, and 19% rate of memory deficits.

Conclusions

Outcomes for MBM patients receiving WBRT indicate that WBRT remains an effective treatment strategy to control intracranial disease. Treatment-related toxicities such as intralesional hemorrhage, necrosis, or neurocognitive side effects are limited. With continued innovations in WBRT technique and systemic therapy development, MBM outcomes may continue to improve. Further trials should evaluate the role of WBRT in the modern context.
Analysis of patient access to orphan drugs in Turkey
Authors: Koçkaya Güvenç , Atalay Sibel , Oğuzhan Gülpembe , Kurnaz Mustafa , Ökçün Selin , Sar Gedik Çiğdem , Şaylan Mete , Şencan Nazlı
Published: February 2021

Background

Rare diseases are life-threatening, serious, and chronic conditions that require complex care and have a low prevalence. An estimated one in 15 people worldwide are affected by rare diseases. This study aims to analyze the accessibility, reimbursement status, licensed status, and Anatomical Therapeutic Chemical (ATC) codes of drugs that the European Medicines Agency (EMA) in Turkey considers to be “orphan” pharmaceuticals.

Methods

The drugs included in this analysis were obtained from the list of orphan drugs published by the EMA. Orphan drugs’ accessibility and licensing status in Turkey were obtained from the Health Implementation Communiqué published by the Social Security Institution (SGK) and the List of Abroad Active Substance and List of Licensed Products published by the Turkey Pharmaceuticals and Medical Devices Agency (TİTCK). Descriptive analysis was applied to determine the accessibility status of orphan drugs identified by the EMA in Turkey.

Results

Based on the EMA, 105 pharmaceuticals were approved with “orphan drug” status except for drugs that have lost orphan drug status, decommissioned in the European Union and withdrawn from the European Community Register by January 2020. Of the 105 rare drugs on the EMA list, 34 were inaccessible in Turkey. Of the 71 available drugs, 23 (32%) were licensed and 48 (68%) were unlicensed in Turkey. 17 (74%) of licensed products and 17 (35%) of unlicensed products were covered by reimbursement. When orphan drugs’ ATC codes were examined, the most common ATC group was found to be “L—Antineoplastic and Immunomodulatory” agents.

Conclusion

An orphan drug incentive policy is very important to ensure early access to the drugs used to treat rare diseases. Considering the capacity and prices for orphan drugs in Turkey, it can be said that many patients with rare diseases have difficulty in their treatment. It is obvious that such a policy must prepare for the regulation of orphan drugs in Turkey.
Novel bone repairing scaffold consisting of bone morphogenetic Protein-2 and human Beta Defensin-3
Authors: He Wei , Wei Daixu , Zhang Jun , Huang Xiaonan , He Da , Liu Bo , Wang Qilong , Liu Mingming , Liu Ling , Liu Yajun , Tian Wei
Published: February 2021

Background

Synthetic biomaterials assist in modulating the vascular response in an injured bone by serving as delivery vehicles of pro-angiogenic molecules to the site of injury or by serving as mimetic platforms which offer support to cell growth and proliferation.

Methods

This study applied natural phospholipid modified protein technologies together with low temperature three-dimensional printing technology to develop a new model of three-dimensional artificial bone scaffold for potential use in repairing body injuries. The focus was to create a porous structure (PS) scaffold of two components, Bone Morphogenetic Protein-2 and Human Beta Defensin-3 (BMP2 and hBD3), which can synchronously realize directional bone induction, angiogenesis and postoperative antibacterial effects. BMP2 induces osteogenesis, whereas hBD3 is antibacterial.

Results

Our data showed that in the BMP2-hBD3-PS or hBD3-PS scaffolds, BMP2 had a slow-release rate of about 40% in 30 days, ensuring that BMP2 could penetrate into stem cells for osteogenic differentiation for a long time. The scaffolds promoted cell growth when in combination with BMP2, thus showing its importance in promoting cell growth. Alkaline Phosphatase (ALP) staining showed that the ALP content of BMP2-hBD3-PS and BMP2-PS had a significant increase in samples that contained BMP2, thus showing that these scaffolds promoted osteogenic differentiation. In all the constructs that had hBD3, they displayed antibacterial properties with hBD3, having a slow release of about 35% in 30 days, thus ensuring they provided protection.

Conclusion

Based on this study, the 3D printed BMP2 scaffolds show a great potential for the development of biodegradable bone implants.

Level of evidence

Level II, experimental comparative design.
2-Pentadecyl-2-oxazoline ameliorates memory impairment and depression-like behaviour in neuropathic mice: possible role of adrenergic alpha2- and H3 histamine autoreceptors
Authors: Boccella Serena , Guida Francesca , Iannotta Monica , Iannotti Fabio Arturo , Infantino Rosmara , Ricciardi Flavia , Cristiano Claudia , Vitale Rosa Maria , Amodeo Pietro , Marabese Ida , Belardo Carmela , de Novellis Vito , Paino Salvatore , Palazzo Enza , Calignano Antonio , Di Marzo Vincenzo , Maione Sabatino , Luongo Livio
Published: February 2021

Neuropathic pain (NP) remains an untreatable disease due to the complex pathophysiology that involves the whole pain neuraxis including the forebrain. Sensory dysfunctions such as allodynia and hyperalgesia are only part of the symptoms associated with neuropathic pain that extend to memory and affectivity deficits. The development of multi-target molecules might be a promising therapeutic strategy against the symptoms associated with NP. 2-pentadecyl-2-oxazoline (PEA-OXA) is a plant-derived agent, which has shown effectiveness against chronic pain and associated neuropsychiatric disorders. The molecular mechanisms by which PEA-OXA exerts its effects are, however, only partially known. In the current study, we show that PEA-OXA, besides being an alpha2 adrenergic receptor antagonist, also acts as a modulator at histamine H3 receptors, and report data on its effects on sensory, affective and cognitive symptoms associated with the spared nerve injury (SNI) model of neuropathic pain in mice. Treatment for 14 days with PEA-OXA after the onset of the symptoms associated with neuropathic pain resulted in the following effects: (i) allodynia was decreased; (ii) affective/cognitive impairment associated with SNI (depression, spatial, and working memories) was counteracted; (iii) long-term potentiation in vivo in the lateral entorhinal cortex-dentate gyrus (perforant pathway, LPP) was ameliorated, (iv) hippocampal glutamate, GABA, histamine, norepinephrine and dopamine altered levels after peripheral nerve injury were reversed, (v) expression level of the TH positive neurons in the Locus Coeruleus were normalized. Thus, a 16-day treatment with PEA-OXA alleviates the sensory, emotional, cognitive, electrophysiological and neurochemical alterations associated with SNI-induced neuropathic pain.
The de novo CACNA1A pathogenic variant Y1384C associated with hemiplegic migraine, early onset cerebellar atrophy and developmental delay leads to a loss of Cav2.1 channel function
Authors: Gandini Maria A. , Souza Ivana A. , Ferron Laurent , Innes A. Micheil , Zamponi Gerald W.
Published: February 2021

CACNA1A pathogenic variants have been linked to several neurological disorders including familial hemiplegic migraine and cerebellar conditions. More recently, de novo variants have been associated with severe early onset developmental encephalopathies. CACNA1A is highly expressed in the central nervous system and encodes the pore-forming CaVα1 subunit of P/Q-type (Cav2.1) calcium channels. We have previously identified a patient with a de novo missense mutation in CACNA1A (p.Y1384C), characterized by hemiplegic migraine, cerebellar atrophy and developmental delay. The mutation is located at the transmembrane S5 segment of the third domain. Functional analysis in two predominant splice variants of the neuronal Cav2.1 channel showed a significant loss of function in current density and changes in gating properties. Moreover, Y1384 variants exhibit differential splice variant-specific effects on recovery from inactivation. Finally, structural analysis revealed structural damage caused by the tyrosine substitution and changes in electrostatic potentials.
Narrative enhancement and cognitive therapy (NECT) to improve social functioning in people with serious mental illness: study protocol for a stepped-wedge cluster randomized controlled trial
Authors: Dubreucq J. , Faraldo M. , Abbes M. , Ycart B. , Richard-Lepouriel H. , Favre S. , Jermann F. , Attal J. , Bakri M. , Cohen T. , Cervello C. , Chereau I. , Cognard C. , De Clercq M. , Douasbin A. , Giordana J. Y. , Giraud-Baro E. , Guillard-Bouhet N. , Legros-Lafarge E. , Polosan M. , Pouchon A. , Rolland M. , Rainteau N. , Roussel C. , Wangermez C. , Yanos P. T. , Lysaker P. H. , Franck N.
Published: February 2021

Background

Self-stigma is highly prevalent in serious mental illness (SMI) and is associated with poorer clinical and functional outcomes. Narrative enhancement and cognitive therapy (NECT) is a group-based intervention combining psychoeducation, cognitive restructuring and story-telling exercises to reduce self-stigma and its impact on recovery-related outcomes. Despite evidence of its effectiveness on self-stigma in schizophrenia-related disorders, it is unclear whether NECT can impact social functioning.

Methods

This is a 12-centre stepped-wedge cluster randomized controlled trial of NECT effectiveness on social functioning in SMI, compared to treatment as usual. One hundred and twenty participants diagnosed with schizophrenia, bipolar disorder or borderline personality disorder will be recruited across the 12 sites. The 12 centres participating to the study will be randomized into two groups: one group (group 1) receiving the intervention at the beginning of the study (T0) and one group (group 2) being a control group for the first 6 months and receiving the intervention after (T1). Outcomes will be compared in both groups at T0 and T1, and 6-month and 12-month outcomes for groups 1 and 2 will be measured without a control group at T2 (to evaluate the stability of the effects over time). Evaluations will be conducted by assessors blind to treatment allocation. The primary outcome is personal and social performance compared across randomization groups. Secondary outcomes include self-stigma, self-esteem, wellbeing, quality of life, illness severity, depressive symptoms and personal recovery.

Discussion

NECT is a promising intervention for reducing self-stigma and improving recovery-related outcomes in SMI. If shown to be effective in this trial, it is likely that NECT will be implemented in psychiatric rehabilitation services with subsequent implications for routine clinical practice.

Trial registration

ClinicalTrials.gov NCT03972735. Trial registration date 31 May 2019.
Poly(4-amino-3-hydroxynaphthalene-1-sulfonic acid) modified glassy carbon electrode for square wave voltammetric determination of amoxicillin in four tablet brands
Authors: Kassa Adane , Amare Meareg
Published: February 2021

Background

Amoxicillin (AMX), which is one of the β-lactam antibiotics used in the treatment of bacterial infections, is known to have a serious mechanism of resistance necessitating continuous monitoring of its level in pharmaceutical and serum samples.

Results

In this study, we presented selective, accurate, and precise square wave voltammetric method based on poly(4-amino-3-hydroxynaphthalene-1-sulfonic acid) modified glassy carbon electrode (poly(AHNSA/GCE)) for determination of amoxicillin in four selected tablet brands. Appearance of a peak in the oxidative scan direction without a peak in the reductive direction of cyclic voltammograms of both bare GCE and poly(AHNSA/GCE) with four folds current and much reduced potential on the modified electrode showed catalytic property of the modifier towards oxidation of AMX. While cyclic voltammetric studies of effect of scan rate showed predominantly diffusion controlled oxidation of AMX with one electron participation, effect of pH revealed participation of protons and electrons in a 1:1 ratio. The square wave voltammetric peak current response of the modified electrode for AMX showed linear dependence on the concentration of the spiked standard AMX in the range 10–150 µmol L−1 with 9.9 nmol L−1 LOD. The AMX content of the studied tablet brands were found in the range 97.84–100.78% of the labeled value. Spike recovery results of 99.6–100.5%, and interference recovery results of 95.4–100.8% AMX in the presence of 50–200% of ampicillin and cloxicillin validated the applicability of the method for determination of amoxicillin in tablet formulation.

Conclusion

In contrast to the previously reported works on determination of amoxicillin, the present method showed an excellent performance making it a potential method for determination of amoxicillin in real samples including serum samples.
Histone crotonylation-centric gene regulation
Authors: Li Kun , Wang Ziqiang
Published: February 2021

Histone crotonylation is a recently described post-translational modification that occurs at multiple identified histone lysine crotonylation sites. An increasing number of studies have demonstrated that histone crotonylation at DNA regulatory elements plays an important role in the activation of gene transcription. However, among others, we have shown that elevated cellular crotonylation levels result in the inhibition of endocytosis-related gene expression and pro-growth gene expression, implicating the complexity of histone crotonylation in gene regulation. Therefore, it is important to understand how histone crotonylation is regulated and how it, in turn, regulates the expression of its target genes. In this review, we summarize the regulatory factors that control histone crotonylation and discuss the role of different histone crotonylation sites in regulating gene expression, while providing novel insights into the central role of histone crotonylation in gene regulation.