Ninety-one percent of global Human Immunodeficiency Virus (HIV) infection in children occurs in sub-Saharan Africa. Provider Initiated Testing and Counselling (PITC) Strategy is a means of reducing missed opportunities for HIV exposed or infected children. The present study determined the prevalence of HIV infection using PITC Strategy among children seen at the Paediatric Emergency Unit of Federal Medical Centre (FMC), Ido-Ekiti, and the possible route of transmission.Cross-sectional study on prevalence of HIV infection using PITC model. 530 new patients whose HIV serostatus were unknown and aged 15 years or below were recruited consecutively and offered HIV testing. Serial algorithm testing for HIV infection using Determine HIV-1/2 and Uni-Gold rapid test kits was adopted. Seropositive patients younger than eighteen months had HIV Deoxyribonucleic Acid Polymerase Chain Reaction (HIV DNA PCR) test for confirmation.Twenty-four (4.5%) of the 530 patients were confirmed to have HIV infection; of whom 19 (79.2%) were less than 18 months of old; with age range of 5 to 156 months. Fifteen (62.5%) of the infected children were females; likewise, the gender specific infection rate was higher (%) among the females compared with (%) among the males. Two of the HIV infected children’s mothers were late, while the remaining 22 mothers (%) were HIV seropositive. Mother-to-child-transmission was the most likely route of transmission in the children.PITC strategy is vital to the early diagnosis and effective control of HIV infection in children. However, this cannot be totally effective if PMTCT is not optimized. Knowledet al.  in a pro [et al.  in an ou [et al. . It is h [et al. . Most stStudy design: the study was a prospective, hospital-based, descriptive cross-sectional study.Study setting: it was carried out at the Paediatric Emergency Unit of FMC, Ido-Ekiti over a period of six months (April-September 2012); a tertiary hospital that serves as a referral centre for the neighbouring towns in Ekiti State and neighbouring Ondo, Osun and Kwara States. It is a 180-bedded hospital that runs general and specialist clinical services in twenty departments. There is an ongoing HIV infection treatment, care and support programme in the hospital. The Paediatric Emergency Unit is a 13-bedded ward with about 100 patients seen monthly (unpublished data).Study participants: the subjects were consecutive new paediatric patients with unknown HIV serostatus, aged 0-15 years who presented in the PEU with any illness. The patients were recruited after signing or thumb-printing an informed consent form by the parents/caregivers. The assent of the patients who were seven years and older was sought by explaining the purpose of the study and details of the sample collection to them in a manner they would understand. Patients with documented HIV status at presentation were excluded from the study. Each patient was recruited once until the desired sample size was attained.Sample size determination: the minimum sample size required for the study was determined using the formula:P = 0.5 (no similar study had been done in the region); d = 0.05. The estimated minimum sample size was 385. However, a total of five hundred and thirty patients were however tested.Data collection: caregivers and patients were given HIV pre-test counselling using WHO guideline on PITC with the choice of “opting out” [ing out” . Counseling out”  was adopThe testing protocol for HIV infection: rapid test kits used for the HIV test were determine and Unigold. The HIV test was carried out and read by the researcher at the side-laboratory attached to the PEU according to the manufacturer’s instructions. Serial algorithm testing for HIV infection was used [was used . The tesPatient management: patients that had HIV infection were immediately enrolled into the paediatric HIV/AIDS treatment, care and support programme at the hospital for full evaluation and treatment as recommended by the National Guidelines for the treatment of Paediatric HIV infection in Nigeria [ Nigeria .Data analysis: the data were entered into a personal microcomputer and analysed using the software, Statistical Package for Social Sciences (SPSS) version 15.0 Inc. Chicago, Illinois-USA. Categorical variables were expressed in proportions, ratios and percentages, while statistical test was done using Chi-square (χ2) test. Statistical significance was set at p-value less than 0.05.Ethical consideration: Institutional Ethical Approval was obtained from the Ethics and Research Committee of FMC, Ido-Ekiti. A written informed consent form detailing the study purpose, benefit, and possible risks to participants and their caregivers was duly signed by each caregiver. In addition, assent was obtained from children aged seven years and above who were in stable clinical condition.A total of 530 patients consisting of 296 (55.8%) males and 234 (44.2%) females participated in the study. The ages of the patients ranged between one day and 180 months, with a median age of 14 months. More than half (59.8%) of their caregivers were in social class III . Twenty-et al. [et al. [The prevalence of HIV infection has been shown to vary with locality and population subgroups. The prevalence of undiagnosed HIV infection among new patients in the present study using PITC Strategy was 4.5%. This was close to the overall national prevalence of 4.1% which also employed PITC Strategy though in a different clinical setting . It was et al.  reported [et al.  in 2001  [et al. .et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [The prevalence rates of paediatric HIV infection in earlier reports from Nigeria were also higher than the prevalence rate in the present study. Akpede et al.  in 1997  [et al.  in 1998  [et al.  reported [et al.  observed [et al.  in their [et al.  however  [et al. , 24, 25. [et al.  found a  [et al.  in Awka  [et al.  in Lagos [et al.  from Lag [et al.  from Awket al., [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [The differences between the prevalence rates in the previous reports and findings in the present study may have also been influenced by the documented HIV prevalence in pregnant women attending antenatal care settings in the various locations since studies have shown that a higher proportion of paediatric HIV infection is acquired through MTCT. The prevalence of HIV infection using PITC Strategy as compared with clinical criteria based-screening tends to be lower because relatively low risk populations are being screened. The role of gender as a risk factor for MTCT of HIV is not clear . The malet al.,  Oniyangi [et al.  and Ogun [et al.  which al [et al. , 28. The [et al.  among pa [et al.  also who [et al. . Althoug [et al.  in 1998  [et al.  Ugochukw [et al.  and Angy [et al.  reportedet al. [et al. [et al. [et al. [et al. [et al. [These findings reflect the importance of maternal HIV infection in paediatric HIV infection in Nigeria and therefore paediatric HIV control measures. It is therefore instructive that twenty-one (95.5%) of the 22 mothers of the patients with HIV infection were newly diagnosed after the primary diagnosis of HIV infection in their children in the present study. The diagnosis of HIV infection in a child usually leads to the diagnosis in a parent with unsuspected HIV infection as it was the case in the present study. Such a discovery would enhance the linking of mothers to HIV infection prevention and treatment programme. While this is complementary in the effort at combating HIV/AIDS in Nigeria, a situation where most infected mothers are detected before or during pregnancy is most desired. The contribution of blood transfusion to the prevalence of HIV infection has been documented , 23, 28.et al.  Ugochukw [et al.  and Angy [et al.  reportedThe prevalence of HIV infection among patients presenting to the Paediatric Emergency Unit of the hospital was high with the predominant route of transmission being MTCT. It is recommended that Provider-Initiated Testing and Counselling Strategy be offered to all children presenting in health facilities; especially immunization clinics, under 5 well child clinics and also other wards that admits children. This will provide opportunity for early diagnosis and treatment in order to reduce the high mortality in children with HIV infection.Provider initiated testing and counselling for HIV leads to significantly higher rates of detection of new cases of HIV infection compared with screening based on clinical suspicion;Maternal to child transmission is the most common route of HIV infection in children;In Nigeria, Ekiti state has one of the least prevalence rates based on the sentinel survey among pregnant women attending ANC clinics in 2010.The prevalence of undiagnosed HIV infection among children presenting in an emergency care setting in Ekiti state, South-western Nigeria is higher than the prevalence rate documented among pregnant women attending ANC clinics in the same study area;All HIV seropositive infants above nine months were confirmed HIV infected using HIV DNA PCR test;Majority of the HIV infected children were less than two years.The authors declare no competing interests.


Gene function, including that of coding and noncoding genes, can be difficult to identify in molecular wet laboratories. Therefore, computational methods, often including machine learning, can be a useful tool to guide and predict function. Although machine learning has been considered as a “black box” in the past, it can be more accurate than simple statistical testing methods. In recent years, deep learning and big data machine learning techniques have developed rapidly and achieved an amazing level of performance in many areas, including image classification and speech recognition. This Research Topic explores the potential for machine learning applied to gene function prediction.We are pleased to see that authors brought the latest machine learning techniques on gene function prediction. Submissions came from an open call for paper, and they were accepted for publication with the assistance of professional referees. Forty-six papers are finally selected from a total of 72 submissions after rigorous reviews. They were presented from different countries and regions, including China, USA, Poland, Taiwan, Korea, Saudi Arabia, India, and so on. According to the topics, we categorize three subtopics for our special issue.Su et al. proposed a novel method called GPSim to effectively deduce the semantic similarity of diseases. Yu et al. constructed a weighted four-layer disease–disease similarity network to characterize the associations at different levels between diseases. Three papers paid attention to miRNA and disease relationship. Qu et al. proposed a novel method to predict miRNA–disease associations based on Locality-constrained Linear Coding. Zhao et al. proposed a novel computational model of SNMFMDA (Symmetric Nonnegative Matrix Factorization for MiRNA-Disease Association prediction) to reveal the relation of miRNA–disease pairs. He et al. proposed an NRLMFMDA (neighborhood regularized logistic matrix factorization method for miRNA–disease association prediction) by integrating miRNA functional similarity, disease semantic similarity, Gaussian interaction profile kernel similarity, and experimental validation of disease–miRNA association. Besides miRNA, there is still a paper on lncRNA–disease relationship prediction. A dual-convolutional neural networks with attention mechanism–based method are presented for predicting the candidate disease lncRNAs .The first part of this special issue discusses the gene and disease relationship. Six papers included in this part are focused on general diseases. These papers propose novel methods to predict disease and gene/miRNA/long noncoding RNA (lncRNA) associations. Liu et al. classified muscle-invasive bladder cancer into two conservative subtypes using miRNA, mRNA, and lncRNA expression data; investigated subtype-related biological pathways; and evaluated the subtype classification performance using machine learning methods. Jiang et al. employed spectral clustering and a novel kernel to predict cancer subtypes. Two papers are focused on breast cancer. Abou Tabl et al. present a hierarchical machine learning system that predicts the 5-year survivability of the patients who went through specific therapy. Li et al. employed machine learning methods to select 54 novel breast cancer oncogenes and proved their findings with GO and KEGG. Three papers researched on other kinds of cancer. Liu et al. found lncRNA LINC00941 as a potential biomarker of gastric cancer. Gao et al. proposed an ensemble strategy to predict prognosis in ovarian cancer. Guo et al. developed rigorous bioinformatics and statistical procedures to identify tumor-infiltrating bacteria associated with colorectal cancer.There are seven papers on cancer and oncogenes. Two papers paid attention to cancer subtypes. Zhuang et al. employed a two-sample Mendelian randomization method to analyze the causal relationships between interleukin 18 (IL-18) plasma levels and type 2 diabetes using IL-18–related SNPs (Single Nucleotide Polymorphism) as genetic instrumental variables. Sun et al. establish a multilevel comparative framework across three insulin target tissues  to provide a better understanding of type 2 diabetes. Zhong et al. identified potential prognostic genes for neuroblastoma. Wang et al. predicted chronic kidney disease susceptibility gene PRKAG2 by comprehensive bioinformatics analysis. Lu et al. employed the Laplacian heat diffusion algorithm to infer novel genes with functions related to uveitis. Li et al. analyzed the blood gene expression signature for osteoarthritis with advanced feature selection methods.Two papers focused on type 2 diabetes and four papers paid attention to other diseases. Oubounyt et al. employed deep learning techniques to predict gene promoter regions. Dao et al. gave a review for detecting DNA replication origins in eukaryotic genomics with machine learning methods. Exons skipping is an important issue in gene structure research. Chen, Feng et al. and Chen, Song et al. analyzed the relationship between histone modifications and exons skipping. Two papers performed researches on RNA secondary structure prediction, which is a classical problem in computational biology. Wang et al. and Zhang et al. employed deep learning to predict RNA secondary structure, especially on pseudoknots.The second part focused on gene structure and function prediction. Four papers were involved in gene elements, and two papers researched RNA structure. Zhang et al. predicted noncoding RNA function with deep learning network. Zhao and Ma employed Multiple Partial Regularized Nonnegative Matrix Factorization for Predicting Ontological Functions of lncRNAs. Deng et al. proposed an integrated model to infer the gene ontology functions of miRNAs The work was supported by the National Key R&D Program of China (2018YFC0910405), the Natural Science Foundation of China (No. 61771331). by integrating multiple data sources. Zou et al. predicted enzyme function with hierarchical multilabel deep learning.Besides gene structure prediction, four papers focused on the gene function prediction, and five papers paid attention to gene identification. Due to the GO- and KEGG-rich knowledge for gene function, researchers would like to pay attention to noncoding RNA function prediction. Han et al. predicted ion channels genes and their types. Chen et al. paid attention to MADS-box gene classification and clustering. Liu et al. predicted gene expression patterns with a generalized linear regression model. Fu et al. identified microRNA genes with sequence and structure information. Qiang et al. predicted RNA N6-methyladenosine sites with machine learning and sequence features.There are also five papers on gene identification, expression pattern prediction, and sites modification. They are all involved with machine learning techniques. Zhu et al. predicted drug–gene interactions with Metapath2vec. Xuan et al. resolve this problem with the latest machine learning technique gradient boosting decision tree. Four papers researched lncRNA–protein interaction prediction. Xie et al. predicted this problem with improved bipartite network recommender algorithm. Zhan et al. combined sequence and evolutionary information on this problem. Zhao et al. employed random walk and neighborhood regularized logistic matrix factorization approach. Dai et al. paid attention to complex features for ncRNA–protein interaction prediction. Three papers are focused on plant researches. Qu et al. found effective sequence features for classifying plant pentatricopeptide repeat proteins. Jiang et al. identified rice yield-related candidate genes by walking on the functional network. Zhang et al. mined Magnaporthe oryzae sRNAs with potential transboundary regulation of rice genes associated with growth and defense through expression profile analysis of the pathogen-infected rice. Three papers paid attention to RNA-seq data analysis. McDermaid et al. proposed a new machine learning–based framework for mapping uncertainty analysis in RNA-seq read alignment and gene expression estimation. Wang et al. gave a systems analysis of the relationships between anemia and ischemic stroke rehabilitation based on RNA-seq data. Niu et al. developed rSeqTU, which is a machine learning–based R package for predicting bacterial transcription units from RNA-seq data.Other researches were categorized as the third part of our special issue. There are 12 papers in total in this part. Two papers are focused on drugs. To conclude, papers in this special issue cover several emerging topics of advanced learning techniques and applications for bioinformatics. We highly hope this special issue can attract concentrated attention in the related fields. We thank the reviewers for their efforts to guarantee the high quality of this special issue. Finally, we thank all the authors who have contributed to this special issue.ZQ wrote the manuscript draft. DM helped to revise the text. AKS gave some helpful suggestions. The work was supported by the National Key R&D Program of China (2018YFC0910405), the Natural Science Foundation of China , Statutory Research funds of Institute of Informatics, Silesian University of Technology, Gliwice, Poland (BK/204/RAU2/2019), and the professorship grant of the Rector of the Silesian University of Technology (02/020/RGPL9/0184).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Acomys(spiny mouse) is incorrectly referred to as a hamster strain. It is a member of therodent family (Muridae). Also, the reference cited for Acomysregeneration was inadvertently modified during publication. Seifert et al. (2012a) wasincorrectly referenced as Seifert et al. (2012b). Finally, a reference for thediscussion on partial versus complete ear closure was accidentally omitted . The main text and references have now been corrected online.In the originally published version of this article, the The authors regret these errors.


This study aimed to determine the prevalence, and relationship between dry eye and glycosylated haemoglobin (HbA1c) among patients with diabetes mellitus.this was a descriptive hospital-based study conducted among patients diagnosed with diabetes mellitus and attending the Diabetic Clinic at a tertiary health facility in Ibadan, south-western Nigeria. Dry eye was assessed using the standardized Ocular Surface Disease Index Questionnaire administered to the eligible respondents on dry eye symptoms. Detailed ocular examination including the tear break-up time (TBUT) and Schirmer I test were carried out and a recent glycosylated haemoglobin value was also obtained.one hundred and eighty-nine Type 2 diabetic patients were studied, with 68.8% female and a mean age of 60.2 ± 10.3 years. The frequency of dry eye among patients was 21.7% . The most commonly reported symptoms of dry eye were “feeling of gritty sensation”  and “blurred vision”  while “discomfort in windy areas”  was the most common environmental trigger. No statistically significant correlation was noted between dry eye and HbA1c , and age dry eye is fairly common among patients with diabetes mellitus with most frequent symptoms being gritty sensation and blurred vision. No significant correlation was noted between dry eye and glycosylated haemoglobin (HbA1c). Dry eye is defined as a multifactorial disease of the tears and ocular surface resulting in symptoms of discomfort, visual disturbance, and tear film instability with potential damage to the ocular surface. It is accompanied by increased osmolarity of the tear film and inflammation of the ocular surface . Many paThis was a descriptive hospital-based study carried out in the ophthalmology and diabetic clinics of a tertiary hospital in Ibadan, Nigeria between December 2014 and January 2015. A sample size of 189 was calculated based on a previously reported prevalence of dry eye among diabetics of 52%,  95% confLastly, the tear break-up time (TBUT) was done followed by Schirmer I test with topical anaesthesia 30 minutes later to avoid any interference of results. TBUT was done by instilling a drop of 2% fluorescein strip wetted with sterile water into the conjunctival sac of each eye. The time interval between the last complete blink and the appearance of a random dark spot on the cornea under the cobalt blue filter of the slit-lamp was recorded with a stopwatch, and the mean of three timings was noted. A value of 10 seconds or less was considered as abnormal . SchirmeData management and analysis: data collected was analyzed using the Statistical Package for Social Sciences (SPSS) software . Summary statistics were presented using frequency tables, charts, means and rates. Chi-square and Fishers exact tests were used for categorical variables. Spearman rank-order correlation co-efficient was used to determine the relationship between dry eye and HbA1c. Level of statistical significance was set at < 5%.One hundred and eighty-nine patients participated in the study of which 59(31.2%) were males,  M: F = . One hunet al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [et al. [The prevalence of dry eye in this study was 21.7% , similar to findings of Kaiserman et al.  (20.6%) et al. , 25 . This is similar to previous studies , 13, 16 67. This et al. . These set al. , 33-35.  [et al.  reported [et al.  and this [et al. . LimitatIn conclusion, dry eye is fairly common among patients with type 2 diabetes mellitus in our black African population with most of the affected patients experiencing the mild form of the disease. No significant correlation was noted between dry eye and glycosylated haemoglobin (HbA1c).Dry eye affects the ocular surface and results in tear film instability;Prevalence of dry eye increases with age and is higher among females;Patients with diabetes mellitus have higher prevalence of dry eye disease.A prevalence value for dry eye was derived for the region which can be used in further studies;No significant gender predilection for dry eye was noted in this study;No significant correlation was also noted between dry eye and glycosylated hemoglobin.


A vast body of research demonstrates that many ecological and evolutionary processes can only be understood from a tri‐trophic viewpoint, that is, one that moves beyond the pairwise interactions of neighbouring trophic levels to consider the emergent features of interactions among multiple trophic levels. Despite its unifying potential, tri‐trophic research has been fragmented, following two distinct paths. One has focused on the population biology and evolutionary ecology of simple food chains of interacting species. The other has focused on bottom‐up and top‐down controls over the distribution of biomass across trophic levels and other ecosystem‐level variables. Here, we propose pathways to bridge these two long‐standing perspectives. We argue that an expanded theory of tri‐trophic interactions (TTIs) can unify our understanding of biological processes across scales and levels of organisation, ranging from species evolution and pairwise interactions to community structure and ecosystem function. To do so requires addressing how community structure and ecosystem function arise as emergent properties of component TTIs, and, in turn, how species traits and TTIs are shaped by the ecosystem processes and the abiotic environment in which they are embedded. We conclude that novel insights will come from applying tri‐trophic theory systematically across all levels of biological organisation. Tri‐trophic research has followed two distinct paths: One has focused on the population biology and evolutionary ecology of simple food chains of interacting species; the other has focused on bottom‐up and top‐down controls over the distribution of biomass across trophic levels and ecosystem‐level variables. Here, we provide a roadmap to bridge these two long‐standing perspectives and identify two key challenges. First, determining whether and how ecosystem‐level TTIs emerge from food chain‐ and community‐level TTIs. Second, determining whether and how ecosystem‐level processes and abiotic factors feedback to shape the species traits that drive TTIs. Ecological and evolutionary outcomes of species interactions can only be fully understood after considering the multi‐trophic setting in which species are embedded. For example, phytophagous insects in terrestrial ecosystems go through periodic outbreaks in North America and Europe, destroying millions of hectares of forest each year . Early research treated pairwise interactions among trophic levels as constants and assumed that multi‐trophic systems could be understood by stringing together these pairwise interactions in an additive fashion  and, in doing so, indirectly influence patterns and the amount of herbivory. These indirect effects alter plant trait evolution, population dynamics, and community structure. Natural enemies may also directly influence plant traits  and this can affect plant relative allocation to direct vs. indirect defences and in turn herbivores.Plants alter herbivore–natural enemy interactions. Plant traits  influence herbivores, which indirectly affects natural enemies. Plants may directly influence natural enemies through the production of cues (volatile organic compounds), rewards (food) or morphological  traits to alter natural enemy behaviours in ways that reduce or enhance herbivory.Herbivores alter plant–natural enemy interactions. Natural enemy indirect effects on plants are contingent on herbivore traits influencing risk of predation or parasitism . Conversely, the expression of plant traits that attract predators or parasitoids is contingent upon the presence, type, and amount of herbivory via plant‐induced responses to damage.Under this perspective, non‐additive effects can be broadly classified into three types of interactions, each of which highlights diverse phenomena and are common to any type of tri‐trophic food chain:Ecosystem perspectiveBottom‐up and top‐down control. Feedbacks between the bottom‐up effect of plant productivity and the top‐down effects of natural enemies, where increasing productivity increases top‐down control though higher natural enemy density but may also reduce top‐down control by extending food chain length, resulting in secondary predators or parasitoids suppressing primary predators or parasitoids.Plant community composition. Increased herbivory following reduction in predation and parasitism (i.e. trophic cascades) leads to changes in plant communities from herbivore tolerant to herbivore resistant species, thus altering plant–herbivore interactions.Herbivore behaviour. Natural enemies induce changes in herbivore behaviours through plastic responses or shifts in species composition that reduce herbivory.Research on TTIs from the ecosystem perspective has considered non‐additive effects in three separate contexts.et al. et al. et al. et al. From an ecological standpoint, TTIs have been studied within the context of indirect interactions mediated by changes in both the density and traits (i.e. plasticity) of trophically intermediate species  have clearly selected for the sophisticated traits used by predators to locate these prey Abrams . Similaret al. et al. et al. et al. et al. et al. et al. Parallel to the species interactions perspective, a separate ‘ecosystem perspective’ on TTIs has also developed. Ecosystem‐level TTIs include processes underlying the distribution of biomass among trophic levels, as well as the direct and indirect effects of TTIs on ecosystem‐level processes . Trophic cascades occur from the top‐down when natural enemies indirectly control plant biomass  and green food webs Allison . These cet al. et al. et al. et al. et al. et al. et al. et al. et al. The search for generalities in how trophic interactions affect ecosystem properties has relied on syntheses and meta‐analyses from multiple systems , but rigorous evolutionary tests are lacking , omnivory and intra‐guild predation (IGP) address non‐additive effects emerging from diverse natural enemy communities  and their consequences for community stability , herbivore guilds (grasshoppers and sap‐feeders), and natural enemy guilds (active hunting and sit‐and‐wait spiders)  because of the challenges of selectively manipulating small‐bodied arthropods  because of the ability to selectively manipulate  large‐bodied herbivores and predators  and evolutionary (micro and macro) effects of abiotic forcing on species traits taking part in component tri‐trophic food chains Fig. , arrow 8et al. et al. et al. et al. et al. sensu McGill et al. et al. Although findings from elevational gradients reveal exciting first steps towards linking spatial variation in abiotic forcing and TTIs, key challenges remain. First, patterns of variation in species traits are likely driven not only by the direct influence of the abiotic environment acting on each trophic level but also by indirect effects acting among trophic levels (Rosenblatt & Schmitz Our understanding of the mechanisms underlying TTIs within and across levels of organisation will be accelerated through multiple new technologies, especially as they are applied in the context of experimental manipulations under field conditions and in combination with novel approaches for analysing large, complex datasets Fig. , arrow 9et al. et al. et al. et al. et al. New technologies in analytical chemistry now allow for increased resolution and sensitivity in sampling of volatile and non‐volatile compounds produced by plants and animals. For example, methods such as untargeted metabolomics analyses (e.g. Clancy et al. et al. et al. New genomic technologies provide insight into the genetic basis of the phenotypes underlying TTIs. Such techniques are revealing the genetic architecture of traits and the processes underlying their evolution (e.g. Dobler et al. et al. et al. et al. et al. et al. et al. Genomics and sequencing techniques are also enabling ecologists to identify groups of microbes or individual taxa of importance to arthropod‐dominated food chains (Pineda et al. In addition to microbe mediation of plant and animal phenotypes, microbial community ecology is also revealing how trophic interactions among microbes can drive ecosystem function (Allison & Martini et al. et al. et al. Finally, advances in remote sensing and ecosystem‐level modelling can help connect TTIs with ecosystem processes. The availability of large databases is increasing not only for remote‐sensed plant diversity and traits (Asner & Martin Knowledge gained from research on TTIs has driven the development of much ecological and evolutionary theory, and empirical work demonstrates their importance for the function of both natural and managed systems. Nevertheless, the lens of tri‐trophic theory has not been applied to all levels of biological organisation. By doing so here, we point out gaps in our understanding and suggest novel ways to form linkages across scales of biological organisation by using TTIs. Our review suggests many novel questions that this proposed programme of research can address, but two key challenges subsume many of these finer points. First, determining whether and how ecosystem‐level TTIs emerge from food chain‐ and community‐level TTIs. Second, determining whether and how ecosystem‐level processes and abiotic factors feedback to shape the species traits that drive TTIs. Addressing these challenges will ultimately unite tri‐trophic perspectives under a single paradigm that guides future research in ecology and evolutionary biology.LAR and AP wrote the manuscript, KM and CB edited the manuscript, all content is based on discussions that took place during a workshop attended by all authors under the initiative of KM and CB. All co‐authors contributed substantially to revisions.No new data are associated with this manuscript.


Prevention of thrombotic disorders such as cardiovascular disease and stroke is an urgent and important task for society. Prevention by suitable diet and exercise is recommended by government guidelines in many countries. In order to get useful and practical results by the recommendations, tests employed to assess the thrombotic status of patients, their quality of diet and their exercise levels have conclusive importance.in vivo wall shear rates are not included in these tests.Assessment of thrombotic status has been performed over the years by quantifying thrombotic factors and by measuring function using anticoagulated blood samples. However, these approaches do not seem to be successful in assessing thrombotic status. This may be due in part to the belief that adding calcium to anticoagulated blood can restore properties of the original native blood and that individual quantification of thrombotic factors can reflect properties of overall multifactorial native (non-anticoagulated) blood. However, this assumption is wrong and needs to change, not least in particular because relevant ex vivo) and thrombosis was measured at various shear rates. The Baumgartner, Sakariassen and colleagues thrombosis tests are triggered by various thrombogenic surfaces, including human arterial subendothelium, human fibrillar collagen and human tissue factor/phospholipids at wall shear rates varying from 100 to 32,000 s-1. Blood is drawn directly from an antecubital vein over the prothrombotic surface at various controlled-wall shear rates, thus avoiding coagulation and platelet activation before reaching the prothrombotic surface [A different approach based on physiological  and biological (evolutionary) ideas was proposed by separate groups in the 1970s. The first group was made up of Baumgartner, Sakariassen and colleagues , and the surface . These tet al. introduced the helium–neon laser-induced in vivo thrombosis system established by Kovacs et al. Subsequently they began research with shear-induced thrombosis/thrombolysis (fibrinolysis) tests both in animals and humans using native blood (ex vivo). Yamamoto and colleagues have analyzed the matching results obtained by ex vivo and in vivo tests in animal experiments. Ex vivo and in vivo results were closely correlated, although with exception in rodents with severe endothelial dysfunction, recommending a simultaneous endothelial function test (flow-mediated vasodilation test) [A thrombus is formed by the interaction between blood and blood vessels under blood flow (Virchow's triad). Yamamoto on test) . No corron test) .et al. observed qualitative differences between fruit and vegetable varieties using shear-induced thrombosis/fibrinolysis ex vivo tests and He–Ne laser-induced thrombosis in vivo test. They demonstrated that the antithrombotic activity of fruits and vegetables varies within the same species, that is, there are varieties with antithrombotic activity, those with prothrombotic activity and those with neither effect [Yamamoto r effect . In healr effect . Furtherr effect . Yamamotr effect . Similarr effect ,17,18.ex vivo Global Thrombosis Test (GTT) is based on the principles of flow chamber techniques, first described by Baumgartner and Sakariassen, and their colleagues [et al. [ex vivo Global Thrombosis Test, could be useful in assessing thrombotic status and bleeding of patients, in developing antithrombotic drugs and diets and in proposing antithrombotic programs utlilizing physical exercise for the prevention of thrombotic disorders.The lleagues ,4. The ulleagues  and in plleagues . Tests ulleagues  and the  [et al. . GTT has [et al. . The slo [et al. . If bloo [et al. . GTT det [et al. . Shear-d


Critical Care [We read with great interest the article by Carlo Custodero et al. recently published in cal Care . The autcal Care , 3, whercal Care  and Robecal Care  indicate


Kraemer MUG, Faria NR, Reiner RC Jr, et al. Spread of yellow fever virus outbreak in Angola and the Democratic Republic of the Congo 2015–16: a modelling study. Lancet Infect Dis 17: 330–382017; —The appendix of this Article has been updated. This correction has been made to the online version as of Feb 15, 2019.


Henipavirus, is classified as a Biosafety Level-4 pathogen based on its high pathogenicity in humans and the lack of available vaccines or therapeutics. Since its initial emergence in 1998 in Malaysia, this virus has become a great threat to domestic animals and humans. Sporadic outbreaks and person-to-person transmission over the past two decades have resulted in hundreds of human fatalities. Epidemiological surveys have shown that NiV is distributed in Asia, Africa, and the South Pacific Ocean, and is transmitted by its natural reservoir, Pteropid bats. Numerous efforts have been made to analyze viral protein function and structure to develop feasible strategies for drug design. Increasing surveillance and preventative measures for the viral infectious disease are urgently needed.Nipah virus (NiV), a zoonotic paramyxovirus belonging to the genus  NiV was named after Kampung Sungai Nipah (Nipah River Village) in Malaysia, where it was first isolated in 1998, before its subsequent spread into Singapore via exported pigs in 1999, leading to the abattoir worker infections  infection was listed as a priority disease posing a public health risk by the World Health Organization (Henipavirus [the other pathogenic member of the genus is Hendra virus (HeV), reviewed in  or an alternative open reading frame (C protein) 2), whichet al.et al.et al.et al.et al.et al.et al.et al.et al.et al.et al.Understanding susceptible hosts and routes for the spread of viral disease raises knowledge to curb epidemics. Bats are the second largest order of mammals after rodents  of NiV could be dated to 1356 . The strains were divided into two lineages , with different clinical features and transmission routes in Bangladesh and Malaysia  to prevent it nonspecific binding to host RNA  surrounded by the viral envelope consists of its genome and the N protein, which is essential for the viral life cycle as a template for RNA-dependent RNA-polymerase (RdRp), composed of polymerase L and a polymerase cofactor P  of the N-terminal domain of P, is characterized by an asymmetric pea-like form composed of three heterodimers. N0 remains an open conformation in the complex due to P-mediated inhibition of the polymerization of N  and is then cleaved by cellular protease into F1 and F2 subunits linked by a disulfide bond. Two heptad-repeat regions (HR1 and HR2) in F1 contribute to the membrane merger  encircling the center in its receptor-unbound status and changes its form in the G/Ephrin-B3 complex. Each blade module in the β-propeller enhanced by a disulfide bond (C181–C601) contains a four-stranded (strands S1–S4) antiparallel β-sheet (Fig.  encircliet al.et al.et al.et al.Knowing the geographic distribution and transmission of a virus is the priority for the control of infection and resolving the structure and function of viral protein is the basis for anti-viral drug development. In this review, we are focusing on these aspects of the NiV. As a huge natural reservoir of viruses, including NiV, bats have been under renewed interest. Bats appear asymptomatic when infected by many viruses and play a pivotal role in viral spillover. Continually emerging and reemerging viruses from bats have been reported. In 2012, a novel rubula-like paramyxovirus from fruit bats was found to be responsible for a series of severe clinical symptoms appearing on a female wildlife biologist who performed a 6-week field exploration in South Sudan and Uganda (Albariño


The aim of this systematic review was to explore safety issues and adverse events arising from PRN prescription and administration. Electronic databases including Scopus, PubMed [including Medline], Embase, Cinahl, Web of Science and ProQuest were systematically searched to retrieve articles published from 2005 to 2017. Selection criteria: we included all randomized controlled trials (RCTs) and studies with comparison groups, comparing PRN prescription and administration with scheduled administration, where safety issues and adverse events were reported. The authors independently assessed titles, abstracts and full-texts of retrieved studies based on inclusion criteria and risk of bias. Results were summarised narratively. The search identified 7699 articles. Title, abstract and full-text appraisals yielded 5 articles. The included studies were RCTs with one exception, a pre-test post-test experimental design. Patient populations, interventions and outcomes varied. Studies compared patient-controlled or routine administration with PRN and one trial assessed the effect of a practice guideline on implementation of PRN administration. More analgesia was administered in the patient-controlled than the PRN arms but pain reduction was similar. However, there was little difference in administration of psychotropic medicines. No differences between patient-controlled and PRN groups were reported for adverse events. The PRN practice guideline improved PRN patient education but non-documentation of PRN administration increased. This systematic review suggests that PRN safety issues and adverse events are an under-researched area of healthcare practice. Variations in the interventions, outcomes and clinical areas make it difficult to judge the overall quality of the evidence. Well-designed RCTs are needed to identify any safety issues and adverse events associated with PRN administration.PRN is the acronym for ‘ The search strategy consisted of the keywords below, based on the authors’ experiences and controlled vocabularies such as the MeSH :pro re nata)” or “as needed” or “as required.”“Drug-Related Side Effects and Adverse Reactions” or “Adverse Drug Event” or “Adverse Drug Reaction” or “Drug Side Effects” or “Drug Toxicity” or “Side Effects of Drugs” or “Toxicity, Drug” and “PRN (pro re nata)” or “as needed” or “as required” and Nurs*.“Drug-Related Side Effects and Adverse Reactions” or “Adverse Drug Event” or “Adverse Drug Reaction” or “Drug Side Effects” or “Drug Toxicity” or “Side Effects of Drugs” or “Toxicity, Drug” and “PRN  independently screened titles and abstracts from the retrieved articles and decided which studies met the inclusion criteria: peer-reviewed RCTs in caring sciences, focus on PRN and published in online scientific journals. Next, two independent review authors (M.V. and S.J.) assessed the full-text of selected articles to ensure that they met the above-mentioned inclusion criteria using the methodological checklist developed by National Institute for Health and Care Excellence (NICE) . In caseTwo review authors (M.V. and S.A.) independently extracted the details of articles included in the review in terms of design, sample, intervention, prescription and administration and outcome measurement.Risk of bias is any error or deviation in the design, study process, analysis and reporting of RCTs, which can cause an underestimation or overestimation of results or inferences . Two autThe heterogeneity of the articles precluded a meta-analysis. Results are presented narratively.Since a meta-analysis could not be performed, no articles were excluded due to missing data and there was no assessment of heterogeneity.We used a theoretical framework of patient safety to accommodate the studies’ heterogeneity in terms of designs, participants and interventions.The authors employed the ‘grading of recommendations assessment, development and evaluation’ (GRADE) criteria ,28 to asThese could not be undertaken, due to inability to pool results.Although no language limitations were applied, all relevant articles were in English. Five articles on the safety and efficacy of PRN prescription and administration are included in this systematic review. The characteristics of the studies are presented in The search identified 7699 articles that could be potentially included in the review. From independent appraisal of the titles and abstracts of the articles by two authors (M.V. and S.J.) deleting duplicates (26 articles) and articles not meeting the inclusion criteria (7650 articles) led to the selection of 23 articles. Reading the full-text of the articles by two authors of this systematic review (M.V. and S.J.) for the inclusion criteria and the selection of RCTs over other study designs led to inclusion of 5 articles. Manual search in the references lists of the included studies identified no more articles. The process of the search is described using the preferred reporting items for systematic reviews and meta-analyses (PRISMA) flowchart in The included studies (n = 5) were published between 2005 and 2015. Two studies ,30 were Three studies ,31,32 weInterventions varied. Three studies considered analgesia ,30,31, oThe outcomes in the selected articles were diverse:Only Chibnall et al.  measuredIn Chibnall et al. , routineMorad et al.  measuredThe adverse effects of paracetamol ,Medication errors associated with PRN prescription and administration ,Neurologic deterioration, excessive sedation, nausea, vomiting, pruritus, insufficient analgesia, and/or respiratory insufficiency ,The adverse effects of the medicines, such as gastrointestinal bleeding or upset, as a secondary outcome ,Safety and ADRs using twelve-lead electrocardiograms at screening, after 8 weeks’ treatment and during the treatment-free follow up period .Diverse adverse events were monitored, including:Eighteen studies were excluded ,44,45,46The risk of bias varied between studies .There were variations in the processes of random sequence generation and concealment among the studies .Chibnall et al.  was blinAttrition is reported in All studies followed their protocols and reported their findings accordingly.Baseline characteristics of participants were similar in all studies. Cross-over design may have minimized the risk of allocation bias in the study of Chibnall et al. .p = 0.01), direct social interactions (p = 0.05) and work-like activity (p = 0.06) than during the placebo phase. They spent less time during the treatment phase engaged in independent self-care (p = 0.02). Emotional wellbeing, agitation, sleeping and independent walking did not differ between study phases (p = 0.80). No other studies reported psychological outcomes.Chibnall et al.  found thIn the cross-over trial by Chibnall et al. , the preBaker et al.  investigMorad et al.  exploredHajimaghsoudi et al.  exploredp = 0.003 and 54.8 vs. 29.9 g/h and p = 0.002).Morad et al.  found paHajimaghsoudi et al.  reportedPark et al.  reportedChibnall et al.  reportedBaker et al.  reportedMorad et al.  reportedHajimaghsoudi et al.  found thPark et al.  reportedpro re nata) prescription and administration in healthcare settings. Few randomized controlled trials compare PRN medication regimens with regular administration of the same drug [The authors aimed to investigate safety issues and adverse events associated with PRN (ame drug . We idename drug . Pain reame drug ,32. The ame drug .The paucity and size of relevant studies, diversity of designs, variations in populations and multiple interventions highlight the incompleteness of the evidence in this systematic review on patient safety and adverse events related to PRN administration. More studies are needed to explore whether PRN prescriptions and the associated transfer of decision-making to nurses or patients and reduced bureaucracy, affects patients’ well-being and quality of care. The safety of PRN prescriptions may depend on appropriate education for nurses ,38,44,46Low sample sizes, difficulties with blinding, absence of information on sampling, randomization and attrition in some trials, variations in the designs, interventions, outcomes and results suggest that the overall quality of evidence is very low. The issues affecting the quality of the included studies differed and mainly stemmed from a lack of detail regarding the methods and interventions in the individual studies. Detailed reporting of the signs and symptoms of ADRs or ‘undesirable effects’ as listed in manufacturers’ literature  is essenWe tried to reduce bias during this review by conducting a thorough literature search using different keywords and databases. The Cochrane Risk of Bias Assessment is provided in A previous systematic review  of PRN mInsufficient evidence for PRN administration and prescription suggests that PRN safety issues and adverse events are under-recognized. The development and implementation of PRN guidelines described in one of the studies  did littWell-designed RCTs of PRN prescription and administration are needed to explore patient safety. The efficacy and effectiveness of PRN with other methods of medication administration and prescription is under-explored but our diverse findings suggest that safety will depend on context, both clinical area and staff preparation. PRN practice guidelines should be developed and evaluated , with a


Plectropomus areolatus) from India and found no evidence to support their findings of alternative reproductive tactics, unique school-spawning involving a single male with multiple females, or inverse size-assortment. The study lacks scientific credibility due to a lack of rigor in the methodology used, misinterpretation of observed behaviors, misinterpretation of the literature, and insufficient data. Their approach led the authors to produce spurious results and profound, invalid conclusions that violate the most basic assumptions of mate choice and sexual selection theory as applied to mating systems in marine fishes.Courtship and spawning behaviors of coral reef fishes are very complex, and sufficient sampling effort and proper methods are required to draw informed conclusions on their mating systems that are grounded in contemporary theories of mate choice and sexual selection. We reviewed the recent study by Karkarey et al. (BMC Ecol 17:10,  Plectropomus areolatus) at a “pristine” site off Bitra, a remote atoll in the northern Lakshadweep archipelago off India. As part of their principal findings, Karkarey et al. [P. areolatus at Bitra showed a habitat-specific, inverse size-assortment in relationship to courtship in which “large males courted small females on the reef slope while small males courted equal-sized or larger females on the shelf.” Both of these reported mating behaviors would appear to violate the most basic assumptions of mate choice and sexual selection theory as applied to marine fishes, and thus the study demanded further scrutiny.In a recent issue in this journal, Karkarey et al.  conductey et al.  describeP. areolatus and other marine fishes and seminal literature on mating systems and sexual selection. Based upon the serious issues contained in the study, which we grouped into five categories described below, we concluded that the study by Karkarey et al. [After careful consideration, we report here that the results of Karkarey et al.  are unsuy et al.  lacks scP. areolatus from within a larger school of females. To support their claim, they provide a photograph in the manuscript  taken fg. 2d in ). The auEach of us has observed the video file numerous times, both in real time and in slow motion, and we can find no plausible evidence of spawning or of any of the behaviors described by Karkarey et al. . In direFurther inspection of the proposed “gamete cloud”  served P. areolatus schools. As early as 1999, Johannes et al. [P. areolatus swimming to, from or within spawning aggregations seem to be the only example of single-sex schooling behavior we know of among groupers within this genus”. Johannes et al. [Karkarey et al.  misrepres et al.  reporteds et al.  also dess et al.  proposeds et al.  but failP. areolatus at the site of their reported research. Descriptions by Johannes et al. [Karkarey et al.  also obss et al.  of inters et al.  and servs et al. ) swimmins et al. ).The sequence of behaviors in the video provided by Karkarey et al.  bears noP. areolatus provide evidence that the species does demonstrate two types of ARTs much like many other coral reef fishes with external fertilization  were highest, they reported that sex ratios were highly biased towards males at the core of the aggregation. The authors of the study reported that “courting behavior …seemed to reflect this shortage of females. Females were often harassed  by several males simultaneously and often fled from them.” In the areas where fish densities were highest, up to 40 males were observed engaging in this behavior. Johannes et al. [Previous studies on iewed by ) but notiewed by . Johanneiewed by  describeiewed by  describes et al.  indicates et al.  noted ths et al.  match ths et al. , 13.P. areolatus at Bitra exhibit a unique spawning tactic involving individual males simultaneously mating with multiple females holds serious implications for sexual selection theory and mating systems of groupers and other marine fishes. Anisogamy  generally leads to situations in which male gametes and individual males are in competition with each other to access and fertilize the eggs produced by females [From a broader perspective, the conclusions of Karkarey et al.  that P.  females –16. This females , 17, 18. females , 19.negatively correlated with population density [While the general conclusion by Karkarey et al.  that “sc density –22. That density , 21, 22, density . TherefoP. areolatus, rendering the principal finding of their study and all associated conclusions regarding the existence, costs, and benefits of alternative reproductive tactics in the species as unsupportable and lacking scientific merit.Contrary to prevailing theories supported by extensive empirical evidence and numerous case studies of coral reef fishes, Karkarey et al.  appear tP. areolatus are most commonly observed on the reef several days prior to spawning. During this time period, oocytes have not progressed to the point of hydration [Karkarey et al.  providedydration , 23, 25.ydration , 25.P. areolatus, the abdomens of females are remarkably distended with hydrated eggs  of actual spawning in ggs Fig. . The proP. areolatus showed “a habitat-specific inverse size-assortment”, in which “large males courted small females on the slope, while small males courted equal-sized or larger females on the shelf”. However, their methods and results suffer from a serious flaw: the authors cannot claim the behaviors they observed were actually courtship or representative of inverse-size assortative mating unless they are at least sometimes followed by a spawning event. The authors themselves state they never once observed a spawning event between a male–female pair of fish. The only evidence they presented to justify their findings were that these behaviors were observed at spawning aggregations of P. areolatus in previous studies [Karkarey et al.  contend  studies , 12.While it is unclear whether courtship was measured at all, it is never appropriate to measure courtship  as a “benefit” accruing to males, because it is unknown whether these behaviors led to successful spawning or whether these individuals remained in the observation arena until spawning commenced. The authors did not observe mating in either large or small individuals, so it is also unreasonable to draw conclusions about inverse size-assortative mating. Additionally, local sex ratio  contaminates the courtship rate measurements, because the “benefit” is multiplied by the number of nearby females; this leads to an estimate of higher “benefit” for males on the shelf, even though time spent in “courtship” was claimed as the same in both habitats. This is the approach taken by Karkarey et al. , as showIt is implausible to make sound inferences about mating rates, potential mating opportunities, or costs associated with intra-sexual selection when none of the measurements used to generate them were based on verified reproductive activity. The observations by Karkarey et al.  of male–Conducting comprehensive quantitative analyses of fish mating behavior, courtship and mating rates, and related factors requires careful, appropriate design and analyses. These practices can result in novel findings that propel our interest and understanding in these and other organisms; however, rarely do these findings contradict established theory. In these instances, clear, irrefutable evidence is required that is supported by rigorous methodology, observations, and analyses. While Karkarey et al.  should b


Hearing loss is a common sensory disorder that has been a serious concern globally. It is recently estimated that around 466 million people worldwide have disabling hearing loss, including 34 million children, and this number will increase to over 900 million by 2050 (according to WHO report 2018). The majority of hearing disorders occur due to the death of either inner ear hair cells (HCs) or spiral ganglion neurons (SGNs), thus leading to sensorial neural hearing loss (SNHL). SNHL is known as the most common form of hearing disorder that comprises about 85% of all hearing loss cases. This type of damage is induced by a variety of reasons such as inner ear trauma, ischemia, ototoxic drugs, noise exposure, inflammation, viral infections, genetic deficits, autoimmunologic reaction, and aging. SNHL is generally not reversible due to the lack of regeneration capacity of HCs and SGNs. However, various recent studies have determined that the HCs and SGNs hold a regenerative potential and it is possible to find the cure for SNHL in the near future. This is supported by the clear understanding of the genetical control and signaling pathways involved in the development of HCs and SGNs and their functions, the regenerative potential of residing adult stem cells, and the development of gene therapy and the clinical trials of new pharmaceutical compounds on damaged cochleae. Last year, we have published the first special issue of “Hearing Loss: Reestablish the Neural Plasticity in Regenerated Spiral Ganglion Neurons and Sensory Hair Cells”, and this year in this second special issue, we are presenting a new series of articles to report the most recent advances in several major areas as summarized below: HC development, HC damage and protection, HC regeneration, SGN development and protection, inherited hearing loss, and inner ear drug delivery.In Vivo”) for the first time experimentally demonstrate the backward traveling wave theory by measuring the phase spectra of the basilar membrane vibration at multiple longitudinal locations along the cochlea. X. Cheng et al. (“Modulation of Glucose Takeup by Glucose Transport on the Isolated OHCs”) report that glucose is transported into OHCs via glucose transporter 1 and 4, which are mainly expressed on the lateral wall of OHCs and the glucose antagonist and ATP regulate this energy transport mechanism. S. Liu et al.  determine that Pax2, Sox2, and Prox1 have differential and overlapping temporospatial expression patterns during the development of vestibular and auditory sensory organs in mice.A. Chang et al.  report that the early environmental sounds promote functional maturation of HCs. Acoustic environment significantly decreases the ABR threshold, increases prestin expression in outer HCs, and enhance maturation of ribbon synapses in the postnatal mouse cochleae. P. Chen et al.  demonstrate that the microglial-like cells are present in the developing mouse cochlea, and these cells go through the drastic morphological and distributional changes during the postnatal cochlear development. Also, these cells might participate in the maturation and remodeling of the cochlea. F. Chen et al.  explore the pathogenesis of superior semicircular canal dehiscence (SSCD) in the guinea pig model and report that the bony fenestration of the superior semicircular canal mimics the hearing loss pattern of SSCD patients. J. Hong et al.  investigate that the N-methyl-D-aspartate receptors regulate the number and distribution of inner HC ribbon synapses after gentamicin-induced ototoxicity and their inhibition by antagonist minimized the drug-induced ototoxicity, and thus maintain the integrity of ribbon synapses. L. Xia et al. (“Comparison of Acceptable Noise Level Generated Using Different Transducers and Response Modes”) compare the acceptable noise level (ANL) in 20 mandarin subjects with normal hearing. The author obtained ANL through different methods and determined that the ANL in normal hearing listeners may not be affected by different modes of presentation. M. Waqas et al.  provide a brief review about the mechanisms involved in the HC loss after noise-induced trauma and discuss the recent HC protection strategies to prevent and recover hearing function in mammals after noise-induced damage. X. Cheng et al.  report that the combination of electric and acoustic hearing significantly improves the perception of music and Mandarin tones in pediatric cochlear implant patients. X. Ding et al.  determine the association between tinnitus and sudden sensorineural hearing loss (SSNHL). The authors found that tinnitus can be ameliorated by the successful treatment of SSNHL. G. Li et al.  provide a brief review on the autoimmune mechanisms involved in SSHL and discuss the role of immunosuppressive drugs in immune therapy. N. Zhao et al.  report that the age-related hearing loss causes an increase in PnC sensitivity that in turn enhances acoustic startle responses in C57 mice. B. Li et al.  investigate the extent of SNHL at high frequencies influences the ability to recognize compressed speech of lower frequencies in hearing loss patients.M. Tang et al.  provide a comprehensive review to address the current challenges and problems in stem cell transplant-based treatments in the inner ear against deafness and present a critical viewpoint about electrical stimulations as a physical factor to modulate stem cell behavior and promote stem cell therapy to treat hearing loss.J. Li et al.  determine that the type II SGNs participate in the contralateral suppression of the medial olivocochlear reflex after selectively inducing apoptosis in the type I SGNs using ouabain treatment.MYO15A Mutations Identified in One Chinese Family with Autosomal Recessive Nonsyndromic Hearing Loss”) report three MYO15A variants c.3971C>A (p.A1324D), c.4011insA (p.Q1337Qfs∗22), and c.9690+1G>A. These variants are absent in 200 normal controls and cosegregated with hearing disability in this family. X. Wang et al. (“A Novel p.G141R Mutation in ILDR1 Leads to Recessive Nonsyndromic Deafness DFNB42 in Two Chinese Han Families”) identify a novel p.G141R homozygous mutation in ILDR1 gene that may be the genetic cause of deafness in two unrelated Chinese Han families. X. Wu et al.  report that the novel compound heterozygous missense mutation c.4472C>T p.T1491M and c.1973T>C p.V658A in PTPRQ gene is the genetic cause of recessively inherited sensorineural hearing loss in a Chinese family.H. Du et al. (“Identification of Binding Partners of Deafness-Related Protein PDZD7”) determine the eleven novel PDZD7-binding proteins through yeast two-hybrid screening that are expressed in the inner ear. Most of the new PDZD7-binding partners such as TRIM35, CADM1, AMOT, BLZF1, Numb, KCDT10, CCDC27, and TRIP11 have not been reported before and will help to understand the role of PDZD7 in hearing transduction. P. Li et al. (“Knock-In Mice with Myo3a Y137C Mutation Displayed Progressive Hearing Loss and Hair Cell Degeneration in the Inner Ear”) report that Myo3a kinase domain Y137C mutant mice have an elevated hearing threshold, degenerated inner ear HCs, and structural abnormality in HCs stereocilia after 6 months of age, thus Myo3a is essential for maintaining the intact structure of HC and normal hearing function. S. Hu et al. (“Genetic Etiology Study of Ten Chinese Families with Nonsyndromic Hearing Loss”) identify novel pathogenic variants in six Chinese families with a hereditary hearing loss by targeted next generation sequencing. F. Zhang et al.  synthesize a new nanoparticle capsule that can be used as a drug delivery route for the gentamicin transfer at the specific site in the inner ear. The authors also determine the sustained release capacity of gentamicin from the capsule by We believe that the studies included in this second special issue of “Hearing Loss: Reestablish the Neural Plasticity in Regenerated Spiral Ganglion Neurons and Sensory Hair Cells” provide important insights into cochlear physiology and pathology as well as the important progress in technology that can be translated into clinical application of the medical treatment of cochlear damage in SNHL. We wish that this special issue will represent a significant contribution in the effort to achieve effective protection and treatment of hearing loss in the near future.


Starch has been an inexhaustible subject of research for many decades. It is an inexpensive, readily-available material with extensive application in the food and processing industry. Researchers are continually trying to improve its properties by different modification procedures and expand its application. What is mostly applied in this view are their chemical modifications, among which organic acids have recently drawn the greatest attention, particularly with respect to the application of starch in the food industry. Namely, organic acids naturally occur in many edible plants and many of them are generally recognized as safe (GRAS), which make them ideal modification agents for starch intended for the food industry. The aim of this review is to give a short literature overview of the progress made in the research of starch esterification, etherification, cross-linking, and dual modification with organic acids and their derivatives. Numerous original articles regarding a starch modification by chemical, physical and/or enzymatic procedures have been published and starch has been extensively reviewed from different points of views. Novel reviews published within the last five years have mainly dealt with starch digestion and resistant starch ,4,5,6,7,et al. [et al. [In 2012 Kaur et al.  reviewed [et al.  reviewedAlthough starch has been reviewed in many aspects, and even though many reviews dealing with chemical modifications of starch have been published to date, the application of organic acids and their derivatives in starch modification has not been discussed in detail to the authors’ knowledge. This article will give a short review of the research and advancements regarding the application of organic acids and their derivatives in starch modification. Although the authors have tried to focus on the 2002–2015 period of research, some articles published prior to 2002 are also considered, due to their valuable contribution to the issue.Starch acetates are additives approved in the food industry under number E1420. They are commonly produced with acetic acid and acetic anhydride as starch esterification reagents. In addition, vinyl acetate can also be used for esterification.Concerning the reaction of the above-mentioned reagents with starch, part of hydroxyl groups on anhydro-glucose units is substituted with acetyl groups and, consequently, esters (starch acetates) are formed. The number of acetyl groups incorporated into the starch molecule is dependent on the reactant concentration, pH, reaction time, and presence of catalysts.et al. [Bello-Perez et al.  have proetc. [et al. [et al. [vice versa). However, the highest DS (2.934) was obtained with the 1:1 mixture, although total mmols were kept constant (180 mmol) and the lowest DS (1.837) was obtained when only anhydride was used. These results indicate that the reagent type does indeed influence the reaction efficiency.An increase in the reactant concentration positively influences the reaction efficiency. Namely, the reaction of starch with acetic anhydride in an aqueous medium with NaOH as the catalyst, results in a low degree of substitution, averaging from 0.01 to 0.2 [etc. . Babić e [et al.  have obt [et al.  have aceThe appertaining synthesis routes significantly influence starch acetylation, too, e.g., when producing highly-substituted starch acetate, the addition time of activator potassium carbonate is of great importance—it should be added after acetic anhydride is allowed to penetrate into the starch granules .2SO4 [−), which reacts with acetic anhydride to build a starch acetate and NaOAc.The reaction of acetic anhydride with starch favourably yields C-3 and C-6 esters . HoweverDicke  has repoIf acetylation is performed with vinyl acetate as the acetylating agent, C-2 esters are exclusively produced .et al. [Guan et al.  have repet al.  has repoet al. [Reaction time is an important factor for the acetylation efficiency. Han et al.  have revA temperature increase from 25 to 30 °C facilitates diffusion of acetylating agents and starch swelling, which results in higher yields of a substituted product. However, acetylation is an exothermic reaction and a further increase of the temperature would negatively impact the reaction .et al. [However, Chi et al.  have docWater is a commonly used reaction medium for acetylation. A higher water content aids dissociation, diffusion, and adsorption of the esterifying agent, which is favorable to the reaction. However, if the water to starch ratio exceeds 1.06:1, the reaction efficiency is reduced due to side reactions .Since a high water concentration is required to avoid mixing problems in industrial conditions, acetic anhydride hydrolyzes to acetic acid, which reduces the reaction efficiency. To overcome this problem, pyridine and DMSO can be used as solvents . In addiet al. [2 as a “green” solvent for low substituted starch acetate production. They have obtained potato starches substituted with acetic anhydride using NaOH as the catalyst with a range of the DS between 0.01 and 0.46, showing a high potential of the densified CO2 usage as a solvent medium. In addition, Shogren [A possible solution for these problems can be found in the research of Muljana et al.  who have Shogren  has repoSince an acetyl group is much bulkier than a hydroxyl group, it sterically hinders the structural organization of starch chains. Due to the repulsion between starch molecules, water percolation between chains is facilitated . Thus, tet al. [et al. [et al. [et al. [et al. [et al. [Sodhi and Singh  have reget al.  after acet al. . However [et al.  have obs [et al.  for acet [et al.  for acet [et al.  have pos [et al.  have demStarch paste clarity and freeze-thaw stability are increased by starch acetylation ,26. Due etc.  after gelatinization at 90 °C for 45 min . The reaet al. [Biswas et al.  have preet al. [Thermoplastic starch-maleate esters have been prepared by an extrusion, with glycerol as plasticizer, as reported by Raquez et al. . FTIR anet al. [Tay et al.  have preOrganic acids are extensively researched in terms of starch modification. Most studies are dedicated to commonly used acids and their derivatives, such as acetic anhydride, succinic anhydride, and OSA . HoweverThe variety of modified starches can be obtained not only by the selection of starting starch type, but also by the careful selection of modifying agents, catalysts, reaction temperature, and time. Reactions can be facilitated by the combined application of physical modifications, such as extrusion or pre-gelatinization. The complexity of modification is, therefore, high and by changing only one reaction parameter, it is possible to obtain a new product with significantly different properties. This allows much space for further research on reactions of starch with organic acids.In addition, a very important aspect is that some of these modified starches should be additionally researched in order to determine their safety for the consumption. On the other hand, a lot of them can already be characterized as good functional food ingredients or good bio-based packaging materials. Since starch and organic acids are easily available and are low-cost materials, the authors do not doubt that starch modification with organic acids and their derivatives will attract even greater attention in future studies.


Toxoplasma gondii Using CRISPR/CAS9” by Bang Shen et al.  and “Efficient Genome Engineering of Toxoplasma gondii using CRISPR/CAS9” by Saima M. Sidik et al.  made an impact on him by successfully implementing strategies to genetically manipulate T. gondii using CRISPR/CAS9 gene editing technology.Alfredo J. Guerra works in the field of molecular parasitology and structural biology. In this mSphere of Influence article, he reflects on how “Efficient Gene Disruption in Diverse Strains of  Toxoplasma gondii Using CRISPR/CAS9” by Bang Shen et al.  and “Efficient Genome Engineering of Toxoplasma gondii using CRISPR/CAS9” by Saima M. Sidik et al.  made an impact on him by successfully implementing strategies to genetically manipulate T. gondii using CRISPR/CAS9 gene editing technology.Alfredo J. Guerra works in the field of molecular parasitology and structural biology. In this mSphere of Influence article, he reflects on how “Efficient Gene Disruption in Diverse Strains of  Toxoplasma gondii: “Efficient Gene Disruption in Diverse Strains of Toxoplasma gondii Using CRISPR/CAS9” by Shen et al.  pathway but can also be used to tag specific genes and also generate point mutations in a site-specific manner.It is difficult to overstate the extent to which CRISPR/CAS9  gene editing technology has changed the landscape of genome editing. The field of apicomplexan biology is no exception to this trend. As a relative newcomer to the field of molecular parasitology, there are two papers that strongly influenced my current research in n et al.  and “Effk et al. . AlthougT. gondii. Shen et al. adapted a CRISPR/CAS9 system with a single guide RNA (sgRNA) to target genes in T. gondii for efficient gene disruption. Taking advantage of a resistance to fluorodeoxyribose that results upon the deletion of the uracil phophoribosyl transferase (UPRT) gene, Shen et al. site-specifically disrupted the UPRT locus both by the nonhomologous end-joining (NHEJ) pathway as well as via homologous recombination, where the UPRT locus was replaced with a pyrimethamine-resistant dihydrofolate reductase (DHFR). The strength of the paper by Shen et al. lies in showing that this approach can be extended to different strains of T. gondii, thereby expanding the utility of this approach in interrogating the complex biology of this apicomplexan parasite. Similarly, Sidik et al. were able to show the efficient disruption of genes via NHEJ using a CRISPR/CAS9 system with an sgRNA targeting the SAG1 gene. Sidik et al. then took their system a step further and showed that in a strain that is deficient in NHEJ (ΔKU80), it is possible to introduce site-specific point mutations by supplying a repair template harboring the desired mutation. One advantage of this approach is the ability to use relatively short homology arms—typically 40 bp. Sidik et al. leveraged drug resistance that results from the specific point mutations that were introduced to give a simple yet powerful confirmation that the desired mutations were introduced. Finally, Sidik et al. showed that CRISPR/CAS9 can be used to tag endogenous loci with an epitope tag in a ΔKU80 strain by using a repair template harboring the desired tag flanked by relatively short (∼40-bp) homology arms to target the insertion in frame with the gene of interest.Shen et al. and Sidik et al. spearheaded the implementation of CRISPR/CAS9 for gene inactivation, gene tagging, and insertion of point mutations in T. gondii field as a whole. One impressive follow-up study is a genome-wide CRISPR screen that allowed Lourido and coworkers to determine the degree to which specific genes contribute to the fitness of the parasites in human foreskin fibroblast (HFF) culture (β domain of T. gondii perforin-like protein 1 (TgPLP1)  . In that


Micromachines aims to present the most recent research developments in scalable micro/nanopatterning. A total of eight papers are presented, including three review papers and five original research papers. The topics include “top-down” approaches, “bottom-up” approaches, and the combination of “top-down” and “bottom-up” approaches.This is the golden age of scalable micro/nanopatterning, as these methods emerge as an answer to produce industrial-scale nano-objects with a focus on economical sustainability and reliability. The improvement of scalability and reliability in nanomanufacturing is a key step to move nanotechnology advances closer to the customer market. This special issue in The Scalable Nanomanufacturing (SNM) program of the National Science Foundation (NSF) was initiated in 2011, and provides funding support to research and education to accelerate the commercialization of nanoscale inventions. Khershed Cooper (NSF) reviewed the initiation of the SNM program [Hu et al. provided a review of tip-based nanofabrication (TBN) for scalable manufacturing . TBN tecDu et al. provided a review of stencil lithography, which is a scalable and resistless nanomanufacturing technique . The curChauvin et al. reported a simple and scalable fabrication technique of porous gold nanowires based on laser interference lithography and dealloying . The golQuantum-dots (QD) are semiconductor particles and have been widely used in the fields of biomedical sensing, photovoltaic devices, and nano-electronics. Keum et al. introduced a scalable QD patterning technique using shape memory polymer (SMP) . The adhNanoimprint lithography (NIL) is a rapid and high resolution nanomanufacturing technique. Lin et al. reported a scalable, time-efficient, and high resolution fabrication process of silicon oxide . An NIL Hoshian et al. introduced a non-lithographic method to pattern silicon . Inkjet Ou et al. demonstrated antireflective SiC nanospikes with a simple and low-cost process by using metal nanoparticles as an etching mask . The metThe future of scalable micro/nanopatterning is bright. We hope that this special issue will help scientists and engineers to develop more advanced micro/nanopatterning techniques in the near future. We would like to thank all the contributors to this special issue and the reviewers for providing valuable comments to the submitted manuscripts.


Escherichia coli,” Nigam et al. . Of the 42 corresponding genes, 36 have an ACA sequence, the target site for MazF cleavage, <100 nt upstream of their start codon. Based on this observation, Nigam et al. tested whether NA-dependent changes in the expression of a green fluorescent protein (GFP) reporter were affected by the positions of upstream ACA sequences. The major conclusion drawn by Nigam et al. was that the presence of an ACA sequence <100 nt upstream of a start codon is associated with MazF-dependent regulation of expression for the corresponding gene. As described in detail below, none of the data presented by Nigam et al. support this conclusion.In their recent study, “Stress-Induced MazF-Mediated Proteins in m et al.  identifiE. coli, as would be expected for any trinucleotide sequence. ACA sequences were found upstream of 36 of the 42 genes listed in Table 1 of the article by Nigam et al. While Nigam et al. describe this frequency as “remarkable,” it is anything but. In fact, for the 42 genes listed in their Table 1, the frequency of genes with an ACA trinucleotide <100 nt upstream is not significantly higher than that for the set of all E. coli genes  or the control set of 2,807 genes described by Nigam et al. as having a “free region upstream” . Thus, these data do not support the conclusion that upstream ACA sequences contribute to MazF-dependent regulation. Moreover, given that most 5′ untranslated region (UTR) lengths for E. coli genes are <50 nt . Lastly, we note that the notion of a “stress-induced translation machinery” has not withstood careful additional and independent scrutiny by the field (In summary, the data presented by Nigam et al.  do not spression , and 13/he field  and that


The objective of this study was to investigate the effect of selected biopolymers on the rheological properties of surimi. In our paper, we highlight the functional properties and rheological aspects of some starch mixtures used in surimi. However, the influence of some other ingredients, such as cryoprotectants, mannans, and hydroxylpropylmethylcellulose (HPMC), on the rheological properties of surimi is also described. The outcome reveals that storage modulus increased with the addition of higher levels of starch. Moreover, the increasing starch level increased the breaking force, deformation, and gel strength of surimi as a result of the absorption of water by starch granules in the mixture to make the surimi more rigid. On the other hand, the addition of cryoprotectants, mannans, and HPMC improved the rheological properties of surimi. The data obtained in this paper could be beneficial particularly to the scientists who deal with food processing field.  Merluccius productus) and Alaska pollock . In addition, various other species such as bigeye snapper (Priacanthus spp.), treadfin bream (Nemipterus spp.), lizardfish (Saurida spp.), croaker (Pennahia and Johnius spp.), are now also used in the preparation of commercial surimi in southeast Asia; the functional and compositional properties vary depending on the species used [According to Park and Morrissey , surimi ies used .Surimi is a unique and useful seafood analogue because of its gelling properties ,4. SinceStarch is made up of two polysaccharides, amylose and amylopectin . AmyloseThe chemical, physical, and functional properties of starches have been widely studied. Native starches from various botanical sources, such as maize, sweet potato, potato, and rice, have received extensive attention due to the differences in their structural and physico-chemical characteristics ,30,31. Iet al. [et al. [et al. [et al. [Normally, consumers evaluate the final acceptance of surimi-based products by their textural characteristics, which are considered to be most important. When a surimi-based product contains high-quality surimi as a predominant component, the resulting texture of the product tends to be rubbery . Lee et et al.  also claet al. . Lee et  [et al.  noted th [et al. . The lev [et al.  added va [et al. . The add [et al.  reported [et al. . The bot [et al. . Yang an [et al.  studied  [et al.  reportedet al. [During thermal processing of starch-surimi systems, significant changes in rheological properties were observed due to the gelatinization of starch and sol-gel transformation of fish proteins, using various starches ,49. Wu eet al. ,49 reporG′) value for surimi with a higher addition of starch [G′ did not decrease during the gel resolution stage [G′ value was the absorption of water by starch granules in the surimi-starch mixture, resulting in more rigid surimi. Campo-Deaño and Tovar [G′ and loss modulus (G″)) of sticks prepared from Alaska pollock (AP) and Pacific whiting (PW) surimi mixed with starch at concentrations of 7%, 11%, and 15%. The difference of (G′–G″) had two distinct regions from a rheological viewpoint, namely, a linear viscoelastic region in which the G′ value was greater than the value of G″ and a non-linear region in which the value of G′ decreased with increasing stress or strain and the G″ value exhibited a contrasting trend [et al. [σmax) or strain (γmax). Campo-Deaño and Tovar [G*), also increased. The authors also observed that, with identical starch concentrations, the PW samples had lower viscoelastic moduli than the AP samples [In the study of Chen and Huang , surimi f starch . They alon stage . The addnd Tovar  investigng trend . These rng trend  and More [et al. , could bnd Tovar  found th samples . This reet al. [Dosidicus gigas). Two methods were used to prepare surimi: first, protein precipitation at the isoelectric point (type A), and, second, washing with an acid solution (type B). Four % sorbitol + 4% sucrose + 0.5% sodium tripolyphosphate, 4% sorbitol + 4% trehalose + 0.5% sodium tripolyphosphate, and 8% trehalose + 0.5% sodium tripolyphosphate were added to the surimi of type A or B. Viscoelastic parameter studies showed that surimi type A samples had significantly higher viscoelastic moduli than type B samples and were more rigid than type B samples. In type B samples, the influence of different cryoprotectants was not discernible. In contrast, in type A samples, trehalose favored less initial protein aggregation and therefore a more thermorheologically stable structure.A cryoprotectant is usually used to protect against damage of biological tissue from ice formation. Some cryoprotectants function by lowering the glass transition temperature of the solution or material. Many cryoprotectants function by forming hydrogen bonds with biological molecules like water molecules in a system. However, cryprotectants prevent freezing, and, in a glassy phase, a solution can maintain some flexibility. For proper DNA and protein function, hydrogen bonding in aqueous solutions is important. Thus, the biological material retains its native physiological structure, although it is no longer immersed in an aqueous environment because the cryoprotectant replaces the water molecules. During frozen storage, surimi is liable to lose quality due to denaturation and/or aggregation of myofibrillar proteins. Cryoprotectants prevent the protein, especially actomyosin, from denaturing/or aggregating. Thus the addition of cryoprotectant to surimi is needed to maintain its quality. Many compounds, including some low molecular weight sugars and polyols as well as many amino acids, carboxylic acids and polyphosphates were known to display cryoprotective effects in surimi ,54,17,52et al.  comparedDosidicusgigas) made by two methods  and stored frozen at −15 °C for 6 months were investigated by Campo-Deaño et al. [G′ and G′′ moduli, decreased between 45 and 50°C, reflecting an increase in fluidity of the semi-gel [et al. [T > 70 C, G′ continues increasing, whereas G′′ remains almost constant, indicating the formation of a highly elastic myofibrillar protein gel. Moreover, in type A samples, there were some differences among the different cryoprotectants: in the type A sample containing sorbitol + trehalose caused a significant increase in G′ and G′′ moduli after eight weeks of storage of type A surimi throughout the temperature range. Sorbitol + trehalose had a weak cryopreservative effect on surimi A; in this case, the cryoprotectant may have increased the internal mechanical stress produced by freezing-induced dehydration. The rheological properties of squid surimi  during frozen storage at −18 °C and the influence of five levels of KGM  on the textural properties of grass carp surimi gels were investigated by Xiong et al. [p < 0.05) with the KGM concentration. Xiong et al. [et al. [et al. [Aristichthy nobilis surimi. Therefore, KGM could be a potential enhancer of the gel properties in surimi processing. However, adding more than 2% KGM is not recommended because, at this level, the surimi gels could easily become too hard due to the strong hygroscopicity of KGM. Moreover, higher levels of KGM significantly reduced the whiteness of the surimi gels. The authors suggested that for a better surimi texture, the optimum level of KGM was 1%. The effect of KGM on myofibrillar protein from grass carp (g et al. . They obg et al.  also com [et al.  conclude [et al.  also notThe structure of HPMC is presented in et al. [According to the report of Chen , HPMC haet al.  noted thet al.  used a tet al. . On the et al.  used theThe biological origin of starch has an important influence on the physico-chemical and functional properties of starch systems in surimi and its products. The viscoelastic moduli and the gel strength increased with increasing the starch concentration in surimi. The addition of combinations of cryoprotectants improved the rheological properties of surimi. The breaking force and deformation of the surimi gels increased significantly with the increasing addition of konjac glucomannan from 0 to 2% during frozen storage at −18 °C, which could also affect the textural properties of the surimigels. These phenomena could also increase the gel-forming ability and improve the strength and elasticity. HPMC is a useful gelation aid material to improve the flow properties of surimi.


Peste des petits ruminants (PPR) is a highly contagious transboundary animal disease with a severe socio-economic impact on the livestock industry, particularly in poor countries where it is endemic. Full understanding of PPR virus (PPRV) pathobiology and molecular biology is critical for effective control and eradication of the disease. To achieve these goals, establishment of stable reverse genetics systems for PPRV would play a key role. Unfortunately, this powerful technology remains less accessible and poorly documented for PPRV. In this review, we discussed the current status of PPRV reverse genetics as well as the recent innovations and advances in the reverse genetics of other non-segmented negative-sense RNA viruses that could be applicable to PPRV. These strategies may contribute to the improvement of existing techniques and/or the development of new reverse genetics systems for PPRV. Morbillivirus in the family Paramyxoviridae , a member of genus et al.et al.et al.et al.et al.The availability of complete genome sequences from vaccine strains and field isolates for all four lineages of PPRV -mediated suppression of signaling lymphocytic activation molecule (SLAM) receptor lead to reduced PPRV titers  was previously analyzed and the role of RNA-dependent RNA polymerase (RdRp) was determined in attempted reverse genetics involving the N, P, and L proteins as well as the PPRV leader and trailer for minigenome expression  is a potential bottleneck for viable PPRV rescue   after cell transfection, which may be confused with the CPE induced by the rescued virus. Indeed, the CMV promoter was successfully used to rescue NDV, although it was shown to be less efficient in low virulent strains  is one of the longest paramyxoviruses after the recently described novel Feline morbillivirus . Consequently, compared with other morbilliviruses, isolation of a field PPRV strain can be difficult due to the lack of sensitive cell lines or inadequate conditions of transportation and stocking of samples , and Orf virus, usually exhibit poor growth et al.et al.et al.et al.et al.et al.et al. et al.et al.et al.et al.et al.et al.Conventional cell lines that exhibit high performance in growth and propagation of PPRV are rarely available , virus rescue relies on co-transfection into eukaryotic cells with at least four plasmids representing the full-length antigenomic sequence of the virus and helper plasmids  independently cloned downstream of the T7 promoter in the presence of an exogenous T7 RNA polymerase source. Even though rescue efficiency for some viruses with cytoplasmic replication can be improved under the control of Pol I and Pol II, the T7 promoter is being gradually replaced by the CMV promoter, which is directly recognized by eukaryotic RNA polymerase  cloned into one plasmid vector as illustrated in Fig. et al. , b. In tl. et al., b.et al.The single-plasmid system is a helper plasmid-free-based system that may be driven by a T7 or CMV promoter with or without exogenous T7 RNA polymerase Fig. D. In thiet al.et al.et al.et al.et al.et al.Years after the approval of a global strategy for the control and eradication of PPRV, there are still continued reports of new PPRV cases, even in unusual hosts, worldwide (Boussini


Pankhurst et al., Chem. Sci., 2016, DOI: 10.1039/c6sc02912d.Correction for ‘Inner-sphere  In the original manuscript, the name of one of the authors was spelt incorrectly. The correct spelling of the author name ‘Carlos Alvarez Lamsfus’ has now been clarified and the complete, corrected author list is presented herein.The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers.


Hallberg et al. provide a limited literature review on the reversal of type 2 diabetes mellitus (T2DM) . InsulinAnother critique of the Hallberg et al. review is the omission of important limitations in the study by Saslow et al. . In the Regarding another study, Hallberg et al. describe an impressive 78% “reversal” rate of diabetes at one year using a definition that excludes metformin ; howeverUltimately, the review by Hallberg et al. presents an overly enthusiastic narrative – and not systematic – review of VLCDs for the treatment of T2DM that does not reflect the entirety of the current evidence available. The authors omit a discussion of high-carbohydrate diets being used to treat and reverse T2DM and fail to mention key limitations of the studies cited on ketogenic diets. Moreover, the authors’ working definition of “reversal” does not necessarily reflect improvements in the underlying pathophysiology of T2DM, i.e., insulin resistance or carbohydrate intolerance.