Mycoplasma (M.) bovis is an important pathogen of cattle implicated in a broad range of clinical manifestations that adversely impacts livestock production worldwide. In the absence of a safe, effective, commercial vaccine in Europe, reduced susceptibility to reported antimicrobials for this organism has contributed to difficulties in controlling infection. Despite global presence, some countries have only recently experienced outbreaks of this pathogen. In the present study, M. bovis isolates collected in Denmark between 1981 and 2016 were characterized to determine (i) genetic diversity and phylogenetic relationships using whole genome sequencing and various sequence-based typing methods and (ii) patterns of antimicrobial resistance compared to other European isolates. The M. bovis population in Denmark was found to be highly homogeneous genomically and with respect to the antimicrobial resistance profile. Previously dominated by an old genotype shared by many other countries (ST17 in the PubMLST legacy scheme), a new predominant type represented by ST94-adh1 has emerged. The same clone is also found in Sweden and Finland, where M. bovis introduction is more recent. Although retrieved from the Netherlands, it appears absent from France, two countries with a long history of M. bovis infection where the M. bovis population is more diverse. Mycoplasma (M.) bovis is one of the currently described more than 130 species of the bacterial genus Mycoplasma and one of the major causative agents of bovine mycoplasmosis [https://www.mpi.govt.nz/protection-and-response/mycoplasma-bovis/) and Finland [M. bovis disease, prevention, and control still remain [M. bovis in Europe. Moreover, few studies on the performance of the vaccines currently licensed in the USA exist .lasmosis  and sinclasmosis . The rec Finland  promptediews see ), while inolones . HoweverM. bovis have been conducted for the purpose of phylogenetic typing or characterization of antimicrobial resistance determinants, but these have essentially focused on national populations [Mycoplasma genus is that only circa 895 genomes, mostly incomplete, are currently available in the NCBI assembly database , which is remarkably few compared to other bacterial genera. Specifically, for M. bovis, there are currently only 269 genome assemblies available. Most of them are incomplete, including 175 derived from surveillance projects. Of the 29 fully assembled genomes, eight represent isolates from China, 16 are from a recent study in Canada, one is the species type strain isolated in the USA (PG45T), and only one is of European origin, JF4278, from Switzerland [Several whole genome sequencing (WGS) studies dedicated to ulations ,9,10,11.ulations . One isstzerland ,15,16,17M. bovis was isolated for the first time in 1981, from a pneumonic calf, and by 1984, the first cases of M. bovis mastitis had been diagnosed [M. bovis pneumonia in Danish cattle has increased and was rapidly considered a threat for dairy production. However, studies on prevalence and isolate diversity are scarce in Denmark [M. bovis infections with severe arthritis and diffusely swollen legs in addition to mastitis and other clinical signs in both adult and young dairy cattle [M. bovis despite the lack of recent publicly available data [M. bovis strain population. Extending our understanding of the evolutionary history and population dynamics of M. bovis to the isolates present in Nordic European countries is essential for the development of appropriate and widely applicable control measures and diagnostic tools adapted to the disease epidemiology.In Denmark, iagnosed . From th Denmark ,21. At ty cattle . Finlandble data ,23. In cble data ,25, wherM. bovis clinical isolates collected in Denmark between 1981 and 2016, including several from the aforementioned 2011 outbreak, compared to other isolates representative of the epidemic situation in other Nordic countries  or other European countries . For that purpose, the genomes of the individual isolates were sequenced and compared taking account of their individual history . We also included four high quality genomes of representative strains as references. Genome-wide methods were compared to the previously established, as well as newly proposed, multilocus sequence typing (MLST) schemes [M. bovis study population was assessed in terms of susceptibility to the commonly used veterinary antibiotics in order to assess the potential for antibiotic resistance lineages in Nordic European countries, from where data are scarce.The present study aimed at analyzing the genetic relatedness and evolution of 73  schemes ,27,28 toM. bovis isolates were sequenced, of which 73 were from Denmark , a median coverage of 126× (range 54–927) and a median number of contigs of 99. The median number of coding sequences (CDS) was 775 (see T and the old isolate from Sweden (SVA-B). Strain F9160 from France was the only exception of a recent strain grouped among the old isolates. Most of the isolates from 2011 onwards were clustered into one main branch with three distinct lineages. The first lineage was highly homogeneous  and contained all recent Danish isolates together with the more recent Finnish and Swedish strains, as well as three strains from the Netherlands. The second lineage  was more heterogeneous and comprised French (n = 5) and Dutch (n = 5) isolates, together with Swiss strain JF4278 from 2008 and a single Estonian isolate. The remaining lineage includes five French isolates and one from the Netherlands, together with the Chinese reference strains, which are almost 2% dissimilar from their nearest neighbor (The average nucleotide identity (ANI) calculated using Pyani and the aligned genome sequences revealed a high level of similarity between isolates; in all cases, this was greater than 98%, which is in the upper range of the ANI expected within a bacterial species (>95%) . The isoneighbor . ClusterM. bovis [The cgMLST analysis was based on the alignment of 500 core genes, out of a pan-genome of 1807 genes, using the open reading frames that were more than 95% identical in 99% of the isolates. This alignment was used to infer a phylogenetic tree using the two Chinese reference strains as an outgroup as they were more remote in the ANI analysis . The resM. bovis . The resM. bovis population worldwide.All these isolates distribution patterns are suggestive of (i) a genomic shift in the Danish population structure with time and also of (ii) a potential divergent evolution of isolates after the year 2008, with isolates from Nordic countries clustered in a separate branch and more diversity for the isolates elsewhere in Europe. To further investigate this hypothesis of clonal spread in Nordic countries, MLST analyses were performed to facilitate comparison of an enlarged set of isolates that better represents the global polC subtyping scheme proposed by Becker et al. [M. bovis population structure  ,28, thatr et al.  were usepolC subtypes confirmed the split between old (st1) and more recent (st3) isolates from Denmark but was poorly discriminative for recent strains with only two subtypes identified on the whole set of strains, st2 and st3, st3 being largely dominant . Other minor subtypes  were found mainly in France and the Netherlands, with two recent isolates found to be variants of ST5 differing by one SNP (ST184 and ST185). The potential clonal spread of ST94-adh1 in Denmark and its appearance in other Nordic countries contrasts with the diversity of STs in France and the Netherlands.The distribution of dominant . In contdominant , confirmgle gene . Of notehttps://pubmlst.org/mbovis/ showed evidence of one major cluster gathered around ST5, which could be considered as a majority “common ancestor”  or a greater evolutionary distance . Isolates from all over the world (as seen in entries from PubMLST) share this similarity to ST5, confirming a worldwide distribution. Isolates sharing STs that are more genetically distant from ST5, such as ST14, 18, and 42 (France and the Netherlands in our data set), are also noted to be geographically and temporally widespread, appearing in the US in strains from 1994 and more recently in central Europe and Israel, according to the PubMLST data. As a consequence of the absence of the adh-1 gene, we were unable to further investigate the closest common ancestor to subtype ST94-adh1 subtype. With the new Register scheme  analyses were performed on a subset of 81 isolates for five widely used antimicrobials representing different classes, i.e., enrofloxacin (fluoroquinolone), oxytetracycline (tetracycline), spectinomycin (aminocyclitol), florfenicol (amphenicol), and tilmicosin (macrolides) . These 8As shown in As expected for ruminant mycoplasmas, no antibiotic resistance genes were found by in silico analysis of the genome. Almost all isolates had several 16S and 23S rRNA gene mutations in hotspots consistent with the observed resistance to tetracycline and macrolide (data not shown) ,31,32.M. bovis isolates gathered from Denmark (n = 73) with a further twenty-four representing five other Northern and Western European countries compared. Old (collected in or before the year 2000) and more recent (year 2011 and after) isolates from Denmark and Sweden enabled the investigation of evolutionary trends in the M. bovis genomic background. The different countries participating in this study had contrasting histories with diagnosed M. bovis infections (see Introduction) and different livestock density, Denmark and the Netherlands featuring in the top four in Europe. Taken together, the results of all the different genome or loci-based analyses were mainly in agreement and suggest that the old clonal type closely related to the historical PG45 type strain that circulated in Denmark in the 1980s and 1990s appears to have been replaced by strains belonging to a new dominant genotype. This new dominant Danish clone might have spread to other Nordic countries that are experiencing recent emergence of M. bovis like Finland and Sweden and is also present in the Netherlands. In contrast, a variety of genotypes coexist in France and the Netherlands, some closer to the old isolates group.In this study, various comparative genomic analyses were performed on ninety-seven M. bovis isolates belonging to polC_st1 have not been detected since the year 2000 and have been replaced by polC_st2 and polC_st3 that were both resistant to the most frequently used antimicrobials except fluoroquinolones. This finding is consistent with the spread of a single clone throughout the country that was hypothesized to be related to antimicrobials used as BRD treatment and restrictions to cattle trade as a consequence of the bovine spongiform encephalopathy crises [adh1, ST29 subtype in MLST-1a or MLST-1b, respectively, is clearly dominant in Denmark, Sweden, and Finland . For instance, trade between Denmark and France is extremely limited while being important between Denmark and the Netherlands, which could explain the ST94-adh1/ST29 presence in the Netherlands, but not in France. This observation also warrants further attention. It was also recently demonstrated that the use of contaminated semen during artificial insemination could contribute to M. bovis spread between farms and/or countries and could be at the origin of the clonal spread from Denmark to Finland and Sweden [A similar scenario of “extinction” of a dominant circulating clonal type has already been described in France, where y crises . The MLSountries ), which ountries . The obsach year ,32. In tach year ,35, withach year . Whetheris cases ,37. Catthe globe , supportd Sweden .M. bovis as a species is known to be highly homogeneous. This was confirmed in the current study with a >98% overall nucleotide identity for the whole set of isolates, irrespective of country of origin, year, body site of isolation, or associated clinical manifestation as demonstrated by our ANI analysis. In this context, tools to discriminate different isolates and understand epidemiological trends are of utmost importance, but are not all equivalent. For instance, polC subtyping was insufficiently discriminatory for recent isolates, generating only two subtypes, with the majority of isolates assigned to st3, while st2 isolates appeared restricted to France or the Netherlands. In France, st2 is currently the main clone in the circulation, with st3 constituting a maximum of 20% of the M. bovis population to date [adh-1 gene, one of the 7 loci included in this MLST-1a scheme, limiting the ability of the scheme to effectively subtype such isolates. The adh-1 gene encodes the alcohol dehydrogenase responsible for alcohol oxidation, a characteristic known to vary between Mycoplasma species [adh-1 locus was recently described in different isolates, including Swiss strain JF4278 used here as a reference [adh-1 gene and they belong to three different MLST-subtypes  suggesting this feature may be a consequence of evolutionary genome reduction associated with host adaption described in mycoplasmas, apparently irrespective of the genomic background. The issue of being unable to designate types for MLST-1a was recently addressed in an updated MLST scheme, which uses dnaA instead of adh-1 [https://pubmlst.org/mbovis/). It already comprises 464 entries and was shown to have a better discriminatory index than the “legacy” base or than the MLST scheme from Rosales et al. (MLST-2 here) [dnaA allele alone was poorly discriminatory for the isolates included in our study. Because it considers evolutionary signals originating from the whole genome, either the whole set of CDS (cgMLST) or genome-wide polymorphisms, the WGS-typing approaches are a more appropriate way forward [M. bovis, the existence of the pubmlst.org database remains an asset for the whole scientific community. This study has contributed to the public database (both the genomic and MLST databases) by including a wider range of isolates from Europe. For instance, previously, only 33 isolates in the PubMLST set of 448 isolates studied by Register et al. were of European origin [ to date . The st2 to date   in France (strain F9160) clustered with old isolates, suggesting it could result from a re-emergence of an old clone that had continued to circulate for years in an unusual host or in an environmental reservoir. The F9160 isolate was isolated in 2014 from a calf in a dairy herd located in the Southwest of France, a region mainly known for ovine breeding. In common with recent strains, the polC-st1 (ST17 in MLST-1a) and the emergence of two new subtypes since 2000 has been associated with a loss of susceptibility to antimicrobials [https://www.danmap.org/-/media/arkiv/projekt-sites/danmap/danmap-reports/danmap-2018/danmap_2018.pdf?la=en). Loss of susceptibility to spectinomycin was only noticed in isolates from France (n = 12/14) and the Netherlands (n = 2/9) as noted earlier [M. bovis strains of st3 in France were more prone to acquiring and fixing mutations in the quinolone resistance-determining regions than those belonging to st2 [In France, the apparent disappearance of the crobials . Here, mcrobials . In cont earlier ,41. Simig to st2 . InteresIn contrast, French strain F9160, which shares its genomic background and genetic subtypes with the strains in Denmark isolated prior to 2000, correspondingly features high susceptibility to the tested antimicrobials. This observation further supports the hypothesis that this and other such strains are able to persist over many years in a reservoir without antimicrobial pressure, such as wild fauna or another environment yet to be defined.M. bovis infections in Swedish cattle. In contrast, the isolate from 1996 was from a unique case on a single farm, and between 1996 and 2011, M. bovis was not diagnosed in Sweden . For the whole genome sequence analysis, the isolates were grown in the PPLO medium supplemented as previously described [The 97 isolates included in the study were gathered from the collections available at different participating laboratories, all located in the Northern and Western Europe . Danish escribed , mycoplaescribed  before DPurified genomic DNAs were sequenced on the Illumina MiSeq 2×250PE system at Wageningen Bioveterinary Research , except for strains No. 10419 from Finland, No. 198 from Denmark, and No. 537 from Estonia, for which genomic sequences were already available . SequencMycoplasma-specific genus database from Prokka [The contigs were annotated using the m Prokka  using trPhylogenetic comparisons were made by calculating a whole genome average nucleotide identity (ANI) using Pyani, a Python package, as well as by using a core genome approach  using Roary  and the M. bovis  using the MLST package (https://github.com/tseemann/mlst) to retrieve individual allelic sequences from the genomes. The PubMLST website was developed by Keith and Jolley and is sited at the University of Oxford [M. bovis was developed and updated recently by Register et al.  [adh-1, gltX, gpsA, gyrB, pta2, tdk, tkt) and the revised MLST-1 scheme . Previously undescribed allele sequences for MLST-1a were submitted to the curator for inclusion in the database. In total, there were 1058 entries in the PubMLST database at the date of accession (August 07); 198 of these lacked information on the adh-1 gene, and thus 890 isolates had full profiles for MLST-1a, divided into 185 different subtypes. However, there were six non-assigned complete allelic ST profiles and only a fifth of the 890 isolates originated from Europe. For MLST-1b, 628 entries had full allelic profiles divided between 136 STs, but with only 11% of the isolates from Europe. Because of the absence of the adh-1 gene in many of the strains and the poor representation of European isolates in the PubMLST database, another MLST scheme (MLST-2) developed at APHA  [dnaA; metS, recA, tufA, atpA, rpoD, tkt), with only tkt being common with MLST-1a. According to the literature, 146 isolates divided into 43 STs were retrieved, the isolates from Europe representing 63% of the total. Assigning STs according to MLST-2 involved uploading the different alleles described earlier in two studies from GenBank [All isolates were assigned to subtypes (STs) according to the Multilocus Sequence Typing (MLST) database for f Oxford , while t1b, new) ,28,51. S Agency)  was used GenBank ,37. Thes GenBank ,37. Fromhttps://pubmlst.org for MLST-1a and 1b. For MLST-2, the different STs and corresponding allelic variants were retrieved from the literature, as described previously [Each currently available allelic profile was downloaded from eviously ,41. Seveeviously .polC subtyping as described by Becker et al. [polC genes were extracted with simulate_PCR [For r et al. , polC gelate_PCR  aligned late_PCR  and attrlate_PCR ,31,32. Tlate_PCR , an apprAll comparative genomic data were agglomerated and plotted using R v3.4.4.Escherichia coli strain K-12 substrain MG1655 (GenBank accession number CP014225). The individual positions were then checked for mutations relative to the PG45 type strain.Mutational hotspots associated with resistance according to Sulyok  were invhttps://card.mcmaster.ca/) with > 90% similarity.The presence of resistance genes was investigated in silico by performing a BLAST analysis using the Comprehensive Antibiotic Resistance Database  under Bioproject accession number PRJNA602897.The MIC assays were performed in Anses laboratory of Lyon, France. Eighty-one strains of the 97 sequenced ones were selected , sent on4–105 CFU/mL was spotted onto agar plates containing dilutions of each antimicrobial and grown at 37 °C in 5% CO2 for 5 days. The MIC was defined as the lowest antimicrobial concentration causing 100% inhibition of growth after the incubation period. Only two different concentrations were used per antimicrobial. These concentrations were based on the clinical breakpoints for Pasteurellaceae as defined by the Clinical and Laboratory Standards Institute (CLSI) to be used as interpretative criteria for cattle respiratory disease (CLSI 2015). They were 2 and 8 µg/mL for oxytetracycline and florfenicol; 8 and 32 µg/mL for tilmicosin; 0.25 and 2 µg/mL for enrofloxacin, and 32 and 64 µg/mL for spectinomycin.MICs were determined using the agar dilution method on the modified PPLO agar as previously described . Five anM. bovis isolates collected in Northern and Western Europe over 40 years. Sequence typing, either using the core genome or based on one or more housekeeping genes, showed a high degree of conservation in this pathogen, albeit with a recent appearance of different phylotypes in different countries between 2000 and 2008. Spread of a homogenous, clone in Denmark, which has subsequently emerged in Sweden and Finland, is also suggested. The rationale for emergence and routes of introduction of different phylotypes warrants further study. Together with the possible existence of environmental reservoirs for M. bovis isolates, this opens an important number of potential evolutionary scenarios for M. bovis sub-populations. Recent isolates share the common feature of lowered susceptibility to major antimicrobial families such as macrolides, tetracyclines, and phenicols. The preserved susceptibility to fluoroquinolones and aminoglycosides should draw our attention in the coming years, as it may evolve differently in the different sub-populations. This study highlights the importance of continuing to monitor M. bovis epidemiology within Europe, including other countries such as Germany, Poland, and the United Kingdom, which are important cattle producers and traders.In conclusion, we compared whole genome sequences in order to analyze the genetic relatedness of 97


Lassa fever is a rodent-borne disease caused by Lassa virus (LASV). It causes fever, dizziness, vertigo, fatigue, coughing, diarrhea, internal bleeding and facial edema. The disease has been known in Guinea since 1960 but only anectodical acute cases have been reported to date. In January 2019, a 35-year-old man, a wood merchant from Kissidougou, Forest Guinea, presented himself at several health centers with persistent fever, frequent vomiting and joint pain. He was repeatedly treated for severe malaria, and died three weeks later in Mamou regional hospital. Differential diagnosis identified LASV as the cause of death. No secondary cases were reported. The complete LASV genome was obtained using next-generation sequencing. Phylogenetic analysis showed that this strain, namely the Kissidougou strain, belongs to the clade IV circulating in Guinea and Sierra Leone, and is thought to have emerged some 150 years ago. Due to the similarity of symptoms with malaria, Lassa fever is still a disease that is difficult to recognize and that may remain undiagnosed in health centers in Guinea. Lassa fever is a haemorrhagic fever due to Lassa virus (LASV) that was discovered in 1969 in Nigeria . The dishttps://www.who.int/teams/blueprint). In this article, we describe both the epidemiological and molecular investigation of the Kissidougou case and his contacts.However, Guinea has very few reported acute human cases, and only a few studies have a posteriori described acute Lassa fever cases in the prefectures of Kindia, Faranah, Kissidougou, Guekedou, Macenta and N’Zérékoré in 1992 and 1996–1999 ,13,14. TField investigations were managed by the Agence Nationale de Sécurité Sanitaire (ANSS) in consultation with members of the Ministry of Health, the Gamal-Nasser University of Conakry, the Word Health Organization (WHO), the NGO Médecins sans Frontières (MSF) and the Centre of Disease Control (CDC). A first investigation took place 6–7 February 2019 in the prefecture of Kissidougou where the patient lived. A second investigation took place 6–7 February 2019 in the Mamou prefecture where the patient died . Contact® Filovirus Screen RT-PCR Kit 1.0  on a Smart Cycler II system . The LASV test was carried out using a conventional RT-PCR targeting the glycoprotein (GP) with the primers LVS36+ (5′-ACCGGGGATCCTAGGCATTT-3′) and LVS339- (5′-GTTCTTTGTGCAGGAMAGGGGCATKGTCAT-3′) [® Lassavirus RT-PCR kit 2.0  on the Rotor-Gene Q platform  was performed on the serum sample of the patient in our satellite laboratory in Gueckedou. The LASV-specific RT-PCR kit targets both the S- and L-segments of LASV. The presence of antibodies against HIV-1/HIV-2 and yellow fever virus (YFV) was tested using the INSTI kit  and ELISA  respectively. The presence of the Hepatitis B virus (HBV) was tested using a rapid screening kit . The presence of the Ebola virus (EBOV) and LASV was investigated by reverse transcription–polymerase chain reaction (RT-PCR). Viral RNA was extracted from serum samples using the QIAmp viral RNA kit . In Conakry, the EBOV test was carried out by real-time RT-PCR, using the RealStarTCAT-3′) . The EBOThe serum sample from the patient was dried on a filter paper and sent to the Bernhard Nocht Institute for Tropical Medicine in Hamburg, Germany. Extraction and metagenomic library preparation for next generation sequencing (NGS) on the Illumina platform were performed as described previously . Brieflyhttps://beast.community/programs). The nucleotide sequences of full-length glycoprotein precursor (GPC), nucleoprotein (NP) and polymerase (L) were aligned separately in three data sets including 41 sequences for each. The sequences were chosen to be representative of their clusters after a preliminary analysis done with 64 NP sequences only and elimination of similar or clone sequences. The alignment of nucleotides was realized according to the position of amino acids in the protein alignment. The phylogeny was inferred by the Bayesian Markov Chain Monte Carlo (MCMC) method implemented in the BEAST software, version 1.10.1 . With thOn 7 January 2019, a 35-year-old man consulted a clinic in Kissidougou because of fever accompanied by chills, dizziness, very frequent bilious vomiting and joint pains. Malaria was diagnosed and the patient was treated with paracetamol and quinine. Despite complying with the malaria treatment, symptoms remained unchanged for one week. He returned to the same clinic, which further referred him to the regional hospital located in Kissidougou on 21 January 2019. At the end of the consultation, a diagnosis of malaria and typhoid fever was made. He received a perfusion and a recommendation of hospitalization was made. However, the patient refused to be hospitalized and came back home. After three days of treatment, and feeling that there was no improvement, he went to another health centre. Three days later, he decided to travel to Mamou, where he stayed two days with his family. Noting that there was no improvement in his status, his parents decided to bring him to the regional hospital of Mamou on 28 January 2019, where he was admitted at 2 p.m. to the emergency room at the Epidemic Treatment Centre (CT-EPI). At the time of admission, the symptoms were as follows: fever and chills, dizziness, vomiting, joint pain, diarrhoea and prostration. A diagnosis of severe malaria was given and he received an intensive supportive antimalarial treatment . At 3 p.m., the patient started bleeding with a cough tinged with blood and the epistaxis started. The physician in charge of the intensive care unit raised the suspicion of a viral haemorrhagic fever (VHF) such as the Ebola virus disease. A blood sample was taken and packed for dispatch to the reference laboratory in Conakry. The patient died at 10 p.m. with a picture of toxic-infectious shock and diffuse bleeding. The body was transported to the morgue and transferred to the Red Cross for a dignified and secure burial. n = 7), Mamou (n = 32) and Conakry (n = 2) were identified by the field teams, and further sampled for LASV RT-PCR as per Ministry of Health National Strategy guidelines in the case of VHF suspicion even in the absence of symptoms. None of them were found positive for EBOV or LASV by RT-PCR  and x = Ct), we can estimate the number of copies/mL to be 5.97E+03. The confirmation of a Lassa fever case launched field investigations and contact tracing activities. The investigation in Kissidougou revealed that the patient was an entrepreneur who was a timber trader. Before falling ill, he had spent a few days in Dandou, a village 32 km from Kissidougou, where he frequently went to do business. A total of 41 individuals in contact with the case in Kissidougou (y RT-PCR . Lassa mammarenavirus genus. This clade includes LASV known to circulate in Faranah (Upper Guinea) and in Macenta (Forest Guinea). Trees based on the combined complete glycoprotein (GP) and nucleoprotein (NP)  A and polein (NP) B show thein (NP) .In Guinea, numerous LASV sequences have been largely described in rodents, notably in the region of Faranah (221 sequences), Kindia (6 sequences) and Guékedou (9 sequences) ,11,20,21The strain described here is clearly related to the sequence LIB-807987 observed in 1981 in Liberia. It may be thus speculated that trades, including those related to the timber trade between Forest Guinea and Liberia, and commercial business favour LASV circulation between Kissidougou and Liberia. The case was, indeed, a logger merchant who used to sell wood in Forest Guinea without prior travel history to Liberia. Yet, a clear geographical origin  for this sub-cluster remains speculative due to a lack of information about LIB-807987, as well as a lack of additional sequences. Thus, the name “Kissidougou cluster” is to highlight that it is different to that of the Macenta cluster but that it is also still part of the LASV known to circulate in Guinea. The Macenta cluster also includes three sequences, which, although identified in Liberia, originate from Macenta, Guinea, a town bordering Liberia. For example, the case described in 2018, the sequence of which is referenced under number LIB-LF18040 , was a woman working at SOGUIPAH, near Macenta. She was diagnosed in Ganta, the neighbouring town in Liberia. The other large “Liberia” cluster, composed of sequences identified in, and originating from Lofa, Bong and Nimba counties in Liberia, forms a cluster independent from that of Guinea. Altogether, the description of this new strain allows us to speculate about a “Macenta” and a “Kissidougou” cluster circulating in Forest Guinea. These clusters appear to be more recent than the ones observed in Faranah, Upper Guinea, and in Liberia. This last country is probably the entry point of the virus in the Mano River region .In three weeks, the patient went through two health centres, one in Kissidougou and one in Mamou. Moreover, despite returning to the same health care facility in Kissidougou at a one week interval with worsened symptoms, no suspicion of VHF was considered. Even following his admission in a critical state at the CT-EPI of Mamou, the diagnosis of severe malaria continued to be considered. This indicates that the accurate and rapid clinical diagnosis of VHF in endemic areas is challenging, despite the 20142–016 Ebola virus disease epidemic in the country. This could be improved by providing regular awareness training on VHF case definitions and access to adequate communication tools .On the other hand, the turnaround time for differential diagnosis was short, around two days . This inThis case report shows that early identification of VHF cases remains challenging in remote areas. Regular awareness training may facilitate the implementation of improved field surveillance and early detection. The molecular description supports a new sub-lineage of LASV in Forest Guinea.


Cox proportional hazards regression models were used to calculate the hazard ratio (HR) of PD, and participants were followed for a mean period of 5.0 years. At the end of follow-up, 3,844 adults were diagnosed with PD. After adjusting for potential confounders, participants with anemia had decreased risk of PD compared to adults without anemia (adjusted HR (aHR) 0.894, 95% CI: 0.809–0.989). Furthermore, aHR of PD was 0.698 (95% CI: 0.546–0.891) in moderate to severe anemia and 0.938 (95% CI: 0.843–1.044) in mild anemia. The protective effect of anemia was also more profound in men  than in women . In conclusion, anemia was associated with lower risk of PD, particularly for patients with moderate to severe anemia. Our study suggests that further studies may be needed to clarify the relationship between anemia and PD.Evidence of the association between anemia and risk of PD (Parkinson’s disease) have been accumulating. This study aimed to examine the relationship between anemia and risk of PD in Korean older adults. Korean adults aged 50 years or older who participated in the Korean National Screening Program ( The prevalence of PD was approximately 0.3% in the population aged over 40 years1. With increasing age, the prevalence of PD increases, and as the size of the aging population is rapidly increasing, the total number of PD patients is expected to increase over time3. The socioeconomic burden of PD is steadily increasing due to longevity and is estimated to be at least $14.4 billion a year in the United States5. The medical expense per patient was twice as high for PD patients than for other patients6.Parkinson’s disease (PD) is the second most common neurodegenerative disease worldwide7, and a previous case-control study in the U.S. showed that more patients in the PD group had anemia than patients in the control group8. However, another study found that increase in hemoglobin concentration in peripheral blood from <14 to ≥16 g/dL was associated with increased risk of PD in elderly men, and hemoglobin could serve as the most significant source of peripheral iron9. Concerning iron intake, one study reported that participants with high intake of iron had increased risk of PD, which is consistent with the idea that iron is a risk factor for PD10. However, a meta-analyses of 33 studies showed that iron levels in the blood and cerebrospinal fluid were not different between PD patients and healthy controls11.No effective methods to prevent PD have been instituted since the pathogenesis of PD is not clearly understood. Moreover, results of previous studies concerning risk factors for PD have not been consistent. Therefore, further well-designed studies to reveal the risk factors for PD are a priority to prevent or delay development of PD. Some evidence suggest that anemia is potentially associated with development of PD, and results of previous studies concerning the relationship between anemia and PD have been controversial. A population-based cohort study using claims data in Taiwan demonstrated that newly diagnosed anemia increased the risk of PD8. In a recent cohort study conducted in Taiwan, anemia was not defined by hemoglobin levels but by diagnosis through claims data, which may create diagnostic limitations7. Anemia may have been under-diagnosed because hemoglobin levels of study subjects and severity of anemia were not examined. Also, well-known risk factors for PD, such as smoking and alcohol intake, were not adjusted for in the analysis.To the best of our knowledge, a cohort study investigating the relationship between hemoglobin level and PD risk has not yet been conducted. Although case control studies studying this association have been carried out, there have been no cohort studies, especially in an Asian populationTherefore, we aimed to study the relationship between hemoglobin level and development of PD in this longitudinal cohort study through a nationwide health screening program.The demographics and comorbid diseases of study participants are shown in Table The enrolled study participants were followed for a mean period of 5.0 years. The incidence rates of PD in participants with anemia and without anemia were 0.328 and 0.411 per 1,000 person-years, respectively. The adjusted hazard ratio (aHR) of PD for the participants with anemia was 0.894 : 0.809–0.989), shown in Table When analyzed by anemia severity, lower hemoglobin level was associated with lower risk of PD. The incidence rates of PD in participants with mild anemia and moderate to severe anemia were 0.446 and 0.281 per 1,000 person-years, respectively. The aHR was 0.938 (95% CI: 0.843–1.044) for the mild anemia group and 0.698 (95% CI: 0.546–0.891) for the moderate to severe anemia group  use of a large sample, representing the general population, which identified a sufficient number of patients with PD and 2) use of the definition of anemia based on hemoglobin level.12. Neurons in the substantia nigra tend to undergo apoptosis more frequently in the brains of PD patients than in normal brains13. While the exact mechanism of the degeneration of dopaminergic neurons in PD is not fully elucidated, several possible mechanisms suggesting anemia as a protective factor against PD can be proposed. First, oxidative stress and iron metabolism are hypothesized as possible mechanisms leading to dopaminergic neuronal death in the CNS15. Iron is the most abundant metal in the adult brain16, and it accumulates in neurons of the substantia nigra in PD patients18. Iron is involved in the reactive oxygen species (ROS) system, leading to free radical and oxidative damage to neurons21. This explanation postulates that overproduced reactive oxygen/nitrogen species can cause neurodegeneration21. Free radicals attack cell membrane lipids, leading to lipid peroxidation, which has been found to be increased in both the substantia nigra and red blood cells of PD patients22. Reactive species act as the catalyst for mitochondrial dysfunction, as well as alpha-synuclein misfolding, aggregation, and accumulation24. Moreover, iron itself can cause toxicity to neurons and may be the key factor for the association of anemia and PD. CSF ferritin level was elevated in PD patients and decreased upon treatment with an iron chelator, improving symptoms of PD22. In addition, evidence suggests that free iron boosts alpha synuclein aggregation in the substantia nigra26. Therefore, this study supports the hypothesis that elevated iron level is toxic to neurons in the substantia nigra.The mechanisms underlying the association between anemia and PD is expected to be complicated and not fully understood. The neuronal degeneration in PD is mediated by either apoptosis or necrosis27. More than 99% of alpha-synuclein in human blood exists inside the blood cells, mostly in red blood cells; but alpha-synuclein’s functions in hematopoiesis and immune reactions are unknown29. Erythrocytes secrete alpha-synuclein into plasma, but the significance of this is unknown. The total amount of alpha-synuclein in blood plasma in PD patients was not different from that of those without PD30. The distribution of alpha-synuclein in red blood cells in PD patients is different from control groups31. Although the molecular mechanism of alpha-synuclein regulation in blood cells and neurons is not known, anemia and PD may be associated by alpha-synuclein. Decreased alpha-synuclein in the red blood cells of anemic subjects may result in decreased secreted alpha-synuclein; therefore the amount of alpha-synuclein undergoing pathologic change will be decreased30. Further study is needed to investigate the roles of alpha-synuclein in anemia and PD.Also, alpha-synuclein, one of the most important proteins in the pathogenesis of PD, may be associated with anemia. An insoluble aggregate of alpha-synuclein is a major component of the Lewy body, an intracellular inclusion that is the hallmark of PD32. In our study, low hemoglobin level decreased the risk of PD to a larger degree in men than in women. Iron may play a more important role in the pathogenesis of PD in men, or men may be more vulnerable to iron accumulation in the substantia nigra than women. More research on the etiology of PD is needed to clarify the reasons why (1) PD is more frequent in men and (2) anemia lowers the risk of PD more in men than in women.One notable finding was the sex difference in the association between anemia and PD, with stronger association in males than females. Some epidemiologic studies were conducted to investigate the difference in incidence or prevalence of PD between sexes, and men had higher incidence and prevalence of PD than women33, including a recent large-scale study in Taiwan, which showed a positive association between diagnosis of iron-deficiency anemia (IDA) and PD7. In the Taiwanese study, the claim code of IDA, iron supplementation therapy, was used as exposure. Iron supplementation itself or changes in iron level may have contributed to the development of PD in this population. In addition, unlike the mentioned study, our study accessed demographic and social information of subjects and adjusted for well-known risk factors, such as alcohol consumption and smoking, which were not adjusted for in the mentioned study.Our study is contrary to several previous studies studying the association between anemia and PD35. A European population-based prospective cohort study also demonstrated that current smokers had lower risk of PD than former smokers compared to non-smokers, and a strong dose-response relationship was also found36. Substances in tobacco such as 2,3,6-trimethyl-1,4-naphthoquinone and nicotine itself have been suggested to affect dopamine metabolism and activity in the brain38. Meanwhile, previous research on alcohol and PD risk have been inconsistent. However, a recent meta-analysis suggested an inverse association between drinking and PD, which was supported by case-control studies but not clear in prospective studies39. Another meta-analysis of longitudinal studies also suggested lower PD risk in drinkers compared with non-drinkers40, which was thought to be consistent with the elevating effects of alcohol on urate levels42.Interestingly, our study found that smoking and drinking were associated with lower risk of PD. However, there is numerous evidence on the inverse association between smoking and PD, which is stronger for current than former smokers compared to non-smokersOur study has several limitations. The level of hemoglobin at the time of NHSP examination was used to assess the presence and severity of anemia, and we could not eliminate the factor of iron replacement at the time of examination. For this reason, some of the participants who temporarily had normal hemoglobin level could have been misclassified into the non-anemic group. Thus, the incidence of PD in the non-anemic group may have been underestimated. Moreover, we were unable to clarify the effect of iron replacement therapy itself on the risk of PD. Another limitation is that we were unable to define the physiological onset of anemia, because hemoglobin levels were defined only at inclusion of the study. Because the reliability of PD diagnosis was also one of our concerns, we used the registration of the NHIS, which needs confirmation of healthcare providers to verify the validity of diagnosis. Due to strict classification of PD patients, those who were not registered in this system may have been misclassified to the control group. The probability of misclassification was the same in both groups, which may have led to underestimation of the protective effect of low hemoglobin levels. Another possible limitation is reverse causation. However, results in our sensitivity analysis of 2-year lag time were consistent with our main findings  is a compulsory insurance program that covers most of the healthcare services conducted in Korea. KNHI also operates a National Health Screening Program (NHSP) for members aged 40 years and older. All members undergoing the NHSP are required to complete a self-reported questionnaire and routine examinations, including blood test and chest X-rayth revision, ICD-10. We also extracted health screening data, including lifestyle information , laboratory test results, and physical exam information such as weight and height.This study used National Health Insurance Service (NHIS) data. We extracted medical claims and demographic information from the NHIS data, including age, sex, monthly insurance premium (proxy for household income level), and disease codes using the medical diagnosis of International Classification of Disease, 10n = 12,342,278) between 2009 and 2013. Participants previously diagnosed with PD  and those who had glomerular filtration rate (GFR) less than 30 mL/min  were excluded. Because the objective of this study was to clarify the relationship between anemia, especially iron-related anemia, and PD, we excluded participants whose decreased renal function may affect both iron-metabolism and development of PD46. Study participants with any diseases or medical conditions that increase PD risk were further excluded. We excluded participants who had (1) hereditary and degenerative disease of the central nervous system other than PD  , (2) psychiatric disorders (ICD-10: F00~F99) , (3) traumatic brain injury  , and (4) cerebrovascular diseases (ICD-10: I60~I69) . Participants with diseases or medical conditions likely to cause anemia were also excluded. Participants who were diagnosed with aplastic anemia or hemolytic anemia  , hematologic malignancy, or other blood disorders  (n = 316) were excluded. Finally, study participants with missing data were excluded. As a result, a total of 2,241,119 Korean adults were included in this study  of Samsung Medical Center approved this study (IRB file No. 2017-05-142) and waived the need for informed consent from the study participants as part of the study approval, because of the anonymity of the data retrieved from the NHIS database. All study methods were performed according to the guidelines approved by the Institutional Review Board of Samsung Medical Center, in accordance with the ethical standards in the 1964 Declaration of Helsinki and its later amendments.Anemia was defined through a one-time result obtained at the baseline health screening examinations. We used the anemia definition of the World Health Organization (WHO): hemoglobin level <13 g/dL for men and <12 g/dL for women. Anemia was categorized by severity as mild (Hb ≥ 11 g/dL) and moderate to severe (Hb < 11 g/dL).47. Follow-up time was calculated as time from beginning of the study to the diagnosis of PD, death, or the final day of study.The main outcome of this study was PD incidence. We used ICD-10 codes to identify diagnosis of PD through diagnosis in medical expense claims (ICD-10 code G20) for both outpatient visits and hospitalizations. To ensure the validity of PD diagnosis, we used also used the registry system for rare intractable diseases (RIDs) in the KNHI. Enrollment in this system reduces patient expenses when requiring long-term and expensive treatment, and registration requires confirmation of diagnosis by a physician. Patients with proven PD diagnosis can be registered in this system, and usually neurologists or primary care physicians who have examined the patient issue the required certification of PD diagnosis. Therefore, we identified new PD onset through claims data with ICD-10 code for PD (G20) confirmed by the RID registration code (V124). This method has also been used in previous epidemiologic studies on PD based on the Korean population, and is regarded to be reasonably accurate48. Household income status was divided into two groups of either high or low based on insurance premiums. Smoking status was categorized into never, former, or current smoker. Both alcohol intake and physical activity were classified into non-regular and regular groups. Comorbid conditions that may affect anemia or PD were considered and identified using ICD-10 codes: hypertension (ICD-10: I10-I15), diabetes mellitus (ICD-10: E10-E14), dyslipidemia (ICD-10: E78), and cancer .Physical measurements and serum biochemical parameters were measured by trained staff through standardized protocols. GFR was calculated via the Modification of Diet in Renal Disease (MDRD) equation. Body mass index (BMI) was calculated as weight divided by squared height (kg/m2) and was categorized into three groups according to Asian obesity criteria: underweight (BMI < 18.5 kg/m2), normal to overweight (BMI = 18.5 to 25 kg/m2), and obese (BMI ≥ 25 kg/m2)Chi-square test was performed to compare the baseline characteristics between study participants with or without PD, and with or without anemia. The incidence rate of PD was calculated as number of diagnoses of PD per 1000 person-years with 95% confidence intervals (95% CIs). The Cox proportional hazards model was used to measure the risk of PD according to the presence or severity of anemia. Cox proportional models were adjusted for age, sex, BMI, alcohol consumption, smoking, physical activity, household income status, hypertension, diabetes, dyslipidemia, GFR, and cancer. We conducted sensitivity analysis by performing 1:2 propensity score matching to adjust for the baseline differences, and additionally calculated PD risk for the anemic and non-anemic groups. We also conducted sensitivity analysis for the main outcomes by giving a lag time of 2 years to account for reverse causation. We also used the Fine and Gray competing risk model to account for the risk of death, which is assumed to be different between the groups with and without anemia. All statistical analyses were performed using STATA 14.0 , with statistical significance defined as two-tailed p-value <0.05.Supplementary Information.


Background and Objectives: Skin cancer is one of the most frequently diagnosed malignancies. The main goal of the therapeutic management is total excision with the prevention of recurrence and metastasis. The quality of life of the patients with skin cancer is affected by the morbidity risk, surgery, and cosmetic or functional aspects. The aim of this study was to evaluate the quality of life of patients with skin cancer prior to and post surgical intervention. Material and methods: We performed a prospective study on 247 patients with skin tumors. Quality of life was evaluated through an initial questionnaire that was given to all consenting patients. This was used to determine patients’ mobility, selfcare, normal activities, pain, and despair, using a five-point Likert scale. The general autoperceived health state was also recorded using a 100-point scale. The study included the responses of all patients at hospital admission, after one month of surgery, and after one year of surgery. Results: In patients with squamous cell carcinoma (SCC), the general health state indicator statistically significantly decreased one month after surgery and increased at one-year follow-up. In malignant melanoma (MM) patients, mobility, selfcare, normal activities, and discomfort presented a decrease in values one year after surgery, compared to the values registered at hospital admission. In patients with basal cell carcinoma (BCC), all indicators of quality of life presented an impaired value one year after surgery, after a decreasing trend. The general health state indicator statistically significantly increased one month after surgery and after one year. Conclusions: Surgery is one of the main steps in treating skin cancer. It has a great impact on patients’ quality of life because of pain andthe effect on mobility and normal activities. Skin cancers influence the quality of life of patients both psychologicallyand physically. Skin cancer is considered to be an important public health issues worldwide. The most frequently diagnosed cancers are nonmelanoma skin cancer (NMSC) and cutaneous malignant melanoma (MM) . NMSC reIn recent decades, the cornerstone of the assessment of cancer treatment efficacy was the survival rate, as well as the recurrence or complication rates. Nowadays, the focus is also on the impact of the disease on patients’ quality of life (QoL) . The conThe aim of this study was to evaluate the impact on the QoL of the patients with skin cancer prior to and post surgical intervention using the health-outcome questionnaireEQ-5D-5L (EuroQol 5 dimensions 5 levels).We conducted a prospective study on 247 patients admitted in the Plastic Surgery Department of the Clinical Emergency Hospital “Prof. Dr. Agrippa Ionescu,” Bucharest, Romania, with skin cancer, over a period of 24 months (January 2017 to December 2018). The inclusion criteria were age >18 years, and a current diagnosis of BCC, SCC, MM, or AK, which was considered a precancerous lesion. The preoperative data comprised demographic information, smoking history, and comorbidities. The histopathological details of the tumor and adjuvant therapies were registered. All patients filled out the EQ-5D-5L questionnaire to describe their perceived QoL.t-test and pairedsamples t-test were used. In addition, correlations between different measurements were determined using the Spearman correlation coefficient. Multilinear regression models were constructed with log-transformed variables. All differences and associations were considered statistically significant if the two-sided p-value was less than 0.05 ).Statistical analyses were performed using the SPSS software version 19.0 . For all continuous data, a preliminary statistical analysis was performed in order to test the normality (Smirnov-Kolmogorov test). For nonnormally-distributed variables and noncontinuous data, the comparisons between groups were performed using nonparametric tests , while for the normally-distributed data, Student’s Local ethical agreement and informed consent of the patients were obtained. The number of the document from the Ethical Commission of Clinical Emergency Hospital “Prof. Dr. Agrippa Ionescu” is 1736615, 05.08.2014.The age of the 247 enrolled patients ranged between 26 and 96 years, with amean of 68 ± 13 years. Among the enrolled patients, 142 (58%) were men and 105 (42%) were women. Moreover, 161 (65%) were from urban areas, and 86 (35%) from rural areas. For the statistical analyses, we considered the following subgroups based on the types of skin cancer: SCC group, MM group, BCC group, and AK group .t-test, p < 0.05 for all comparisons). There was no statistically-significant difference between the subgroups regarding gender structure . Regarding the residence area, the structures of the MM group and the AK group were significantly different from the other two groups: the patients in those groups were mostly from urban areas .The mean age of the patients in the MM group was statistically significantly lower than that in all the other groups (p < 0.001), and there was no significant difference between BCC and AKgroups . The distribution of the sun exposure in SCC, BCC, and AK groups was similar, with prevalence in the “frequent” category. The amount of sun exposure in the MM group was particularly different, with equal prevalence in both categories. Skin type distribution among the patients in the four groups presented no significant differences ; skin types 2 and 3 were most prevalent in all groups.We also compared the localization of the tumor, the maximum tumor diameter, solar exposure, and the skin type between the four groups. Because the maximum tumor diameter does not follow a normal distribution, median and range are reported . The maxIn our study, the QoL was assessed using a five-point Likert scale  for mobility, selfcare, normal activities, pain (discomfort), and despair . The genWe analyzed the QoL between the groups as follows.Mobility: There were no significant differences between the groups.t-test, p < 0.001). The values in BCC and AK groups were significantly lower than those in SCC and MM groups. After one year of surgery, the value in the MM group wassignificantly lowerthanthose in the other groups . Selfcare: After one month of surgery, there wasa statistically-significant difference between SCC and MM groups and BCC and AK groups . After one month of surgery, the SCC and MM groups had significantly higher values than those in BCC and AK groups . After one year of surgery, the value in the MM group was still significantly higher than those in the other groups (p < 0.001).Normal activities: At hospital admission, the value in the MM group was significantly lower than those in theother groups . After one year of surgery, the value in the MM group was still significantly higher than those in the other groups (p < 0.001).Pain/discomfort: After one month of surgery, there was a statistically-significant difference between SCC and MM groups and BCC and AK groups . The despair indicator slightly increased after one month of surgery, due to the impact of the histopathological result as well as the scars. The general health state indicator statistically significantly decreased after one month of surgery, and statistically significantly increased after one year . In the SCC group, all indicators of the QoL presented impaired values one year after surgery, after a decreasing trend , with statistically-significant differences . The despair indicator statistically significantly increased after one month of surgery , and remained significantly higher than at hospital admission after one year , due to the impact of being diagnosed with MM. Also, the general health state significantly decreased after one month of surgery , and remained lower after one year, compared to the hospital admission value .In the MM group, the mobility, selfcare, normal activities, and pain/discomfort presented a decrease in values one year after surgery, compared to the values registered at hospital admission, with statistically-significant differences in selfcare . The general health state indicator statistically significantly increased one month after surgery and after one year . In the BCC group, all indicators of the QoL presented impaired values one year after surgery, after a decreasing trend , with statistically-significant differences, except for mobility . The general health state indicator statistically significantly increased after one month of surgery and after one year .In the AK group, all indicators of the QoL presented impaired values (1 or very close to 1) one year after surgery, after a decreasing trend , with statistically-significant differences, except for mobility  on 121 patients demonstrated that the QoL of patients with SCC is more affected in comparison to those with BCC, due to the fact that SCC is a more aggressive lesion that can lead to metastasis, and has a more invasive treatment . A total of 21 patients had AK, with good reports of the QoL after one year. Philipp-Dormston et al. (2018) demonstrated that the progression from AK to SCC is associated with a significant reduction in QoL [In the MM group, the QoL showed a significant decrease, with no additional increase, even after one year. The general health state had a very strong decrease after one month, and remained lower after one year, compared to the hospital admission value. The excision was performed with 0.5 cm at diagnosis, and reexcision was made with 1- or 2-cm oncologic margins, depending on the Breslow depth. The defects were covered with local flaps. Chernyshov et al. 2019) in their QoL report highlighted thatpatients with MM showed an increased physical functioning and bodily pain [ in theirSkin cancer is one of the most common types of malignancies, with a rapidly increasing incidence every year, affecting patients’ QoL. Questionnaires evaluating the QoL are important and should be used by physicians as an outcome measure.


Street food plays an increasingly important role in the nutrition of the inhabitants of European cities. Our study aimed to analyze Polish consumers’ attitudes toward food offered in street food outlets, consumers’ eating out behavior, and the factors that determine their choice of meals from street food vendors. A survey was conducted of 1300 adult respondents who eat street food in Poland. The research enabled a detailed and comprehensive assessment of consumer behavior toward the use of street food outlets, as well as consumer opinions on vendors’ functioning, including hygiene and meals offered. Factors determining the frequency of street food consumption, preferred food types, and factors influencing the use of such outlets were identified. The most important factors were the quality of services and meals, personal preferences and price. Using cluster analysis, consumer profiles based on the types of street food outlets and food preferred were identified. Four main street food consumer preference profiles were identified: ‘burger-enthusiasts’, ‘kebab-enthusiasts’ and ‘ice-cream enthusiasts’, and ‘no specific-oriented consumers’. The Internet and social media were identified as information and promotion channels for this form of gastronomy. Results also revealed Polish consumer behavior and opinions about the food safety of street food in Poland. In summary, in Poland the habits of eating typical Polish homemade dishes is being replaced by eating meals in street food outlets, which can be classified as fast food. Increasing consumer knowledge and awareness of the quality and safety of street food may counteract improper hygiene practices of sellers. Street food terminology defines food and beverages as either ready for direct consumption or not and includes food that is ready-to-eat and food that is minimally processed, prepared and/or sold by vendors and handlers. This is an inexpensive food for all workers of all socio-economic classes and occupations ,2.Street food plays an increasingly important role in the food intake of Europeans . Street Currently, food-trucks are a restaurant marketing strategy in Brazil and in Paris ,9. StreeDue to their locations on the street, the hygienic conditions of preparing and selling food by street vendors are often unacceptable ,17,18,19Due to the poor hygienic conditions and the lack of vendor awareness, foodborne infections from street food occur in both developed and undeveloped countries ,45. ThisStudies of street food establishments primarily focus on food choices and the frequency of using street food ,9,11, deConsumers do not exert pressure on street vendors to improve food preparation conditions . SanitarThree decades ago in Poland, people usually ate homemade meals. Currently, the lifestyle and habits of consumers have changed and influenced foodservice sector development. Among other factors, global trends such as demographic changes (growth of 1–2 members households), economic changes (average household income growth), as well as consumers’ knowledge growth  have all contributed to changes in consumer food consumption Over the two last decades, the amount of money in Polish households allocated to catering services has increased, and the share of food spending and total expenditure doubled, despite the amount being low. The average individual Polish spend on food out of home varies between 26.2 PLN (about 8$ in 2013) to 60.20 PLN (about 16.5$ in 2015), which accounted for 8.5% of their total food budget and 1.9–3.0% (2015–2020) of the total consumption expenditures. For 27 European Union (EU) countries the average share of consumption expenditure equals 7%. The share of household expenditure devoted to catering services was the largest in Ireland (14.4%), Spain (13.0%), Malta (12.6%) and Greece (12.4%), but the lowest in Poland (only 3% in 2017). This indicates a low preference of Poles to eat away from home ,63,64,65Therefore, our research aimed to analyze consumer attitudes toward street food  and eating out behavior, as well as the factors determining the choice of meals in street food outlets.What are the motives (factors) of consumers choosing street food outlets?What consumer profiles can be identified according to the frequency of using street food outlets?How do consumers evaluate street food outlets in terms of food, including food quality, service, and hygiene?The following research questions were formulated:n = 12) within the population of interest. This group was not added to the main research. All problems were identified, and the questionnaire was completed and corrected. The data were collected by the authors using the PAPI (pen-and-paper interview) method.We designed a questionnaire based on available questionnaires ,67 and oEach respondent who agreed to participate in the survey was invited to complete the questionnaire. If necessary, explanations were provided.Everyone, independent of age, using the offer of the street food did not suffer from diseases requiring a special menu offer.Inclusion criteria of respondents for main study were as follows:The exclusion criterion of respondents was people who don’t use street food outlets. The participants in the study were a convenient sample of consumers. They were free to participate in the questionnaire.The questionnaire consisted of two parts . The firp < 0.05.The statistical analysis of the results was performed using Statistica software . The analysis of variance (ANOVA) test and multi-dimensional cluster analysis were used. Significance of differences between the values was determined at a significance level of A multi-dimensional cluster analysis calculation- was performed: hierarchical cluster analysis. The goal of our cluster analysis calculation was to build a tree diagram where the answers given by participants were most similar in a specific cluster. In order to avoid high correlated variables distortions on results, the variance inflation factor (VIF) has been calculated for the possibility of using cluster analysis. For cluster validation statistics we used internal measures for cluster validation, e.g., the matrix and Euclidean distance. The Ward method, as a hierarchical clustering method, was used to create groups, where the variance within the groups is minimized. We used the cluster analysis to determine consumer profiles based on the reasons for using and not using street food outlets and their opinions about these. In the cluster analysis of consumer opinion profiles, features such as price, service, hygiene, promotion, organization, and quality were taken into account, as well as individual component variables.The characteristics of the respondents are presented in n = 1131, 87%) regularly used street food services. The others  used this form of gastronomy occasionally, e.g., at street food festivals, outdoor events, etc. A statistically significant influence of gender (p = 0.00076), age (p = 0.00006), education (p = 0.0031), dwelling place (p = 0.00016), and financial status (p = 0.00001) on using street food outlets was found. This form of gastronomy is significantly more often used by men, people aged 19–30 years, with lower than secondary education, living in the countryside or in cities up to 250,000 of residents, and a ‘bad’ financial status.Many respondents . The highest percentage of respondents visited two or three times a week  and once a week . The remaining respondents chose these facilities less frequently: once a month , once every 2 or 3 months  or less often . The frequency of using this form of gastronomy depended on gender (p = 0.00011), age (p = 0.000001), education (p = 0.00014), dwelling place (p = 0.00006) and financial status (p = 0.000001). Men, and people under the age of 18 years, with the lowest education, living in the countryside or in small towns, and with a ’bad’ financial status used this form of eating outside the home significantly more often, visiting this type of facility every day or several times a week. It is probably the cheapest form of dining away for them when they are at school or at work.The respondents visited street food outlets with varying frequency. This form of catering was used by a small percentage of consumers every day and four or five times a week (p = 0.0010), age (p = 0.000001), education (p = 0.000001), financial situation (p = 0.00001), and dwelling place (p = 0.0014).The choice of street food depended on gender (-p = 0.05) who used street food for three or four times a week;burger-enthusiasts (I), young consumers (aged 19–30 years), mainly men, highly educated, with ‘good’ and ‘very good’ financial status , who used street food for three or four times a week;kebab-enthusiasts (II), young respondents (aged 19–30 years), mainly men , who used street food once a month;ice-cream enthusiasts (III), consumers with various sociodemographic groups , respondents, mainly women, secondary educated, who used street food with different frequency.Based on the cluster analysis, the profiles of preferences of consumers using street food were determined . Four prn = 346, 27%), kebabs  and ice cream  most often. Other consumer profiles were represented by small groups of respondents .Consumers consumed burgers , or social networking sites and internet forums . They also used websites with restaurant reviews , articles on the Internet , articles in the local press and in weeklies , blogs , and others . The following other sources were listed: TV and advertisements such as leaflets, folders, phone applications, radio, advertising on the roads, vlogs, Google Maps, and the website streetfoodpolska.pl. The choice of the source of information about catering establishments depended on gender (p = 0.00001), age (p = 0.000001), education (p = 0.000001), dwelling place (p = 0.0001), and financial situation (p = 0.000001).The respondents use various sources when choosing a catering establishment. Most often these were the opinions of friends, family . Similarly, the most important discouraging factor was the quality factor. It was indicated by over 65% of consumers. Other components of individual factors in both groups are presented in The presence of the ‘preferences and quality’ factor in decisions for choosing a catering establishment is confirmed by the calculations of descriptive statistics, i.e., mean, SD, and medians. In the case of the expectation of the quality of service and the satisfaction of an individual’s own preferences  have indicated ‘quality’  sometimes complained about the quality of the dishes. A small group of respondents  very often reported complaints about catering services. Other respondents almost never  and never  made any complaint about the quality of the dishes. Complaint about food quality depended on the age (p = 0.00001), education (p = 0.000001), dwelling place (p = 0.00181), and financial situation (p = 0.000001) of the respondents. People aged 31–55 years, with vocational and elementary education, living in cities of up to 250,000 inhabitants, and with ’bad’ financial situations made complaints significantly more often.The respondents . They wore jewelry on their hands during work (51%), and they had no or did not change disposable gloves frequently enough (41.5%). They touched their face, hair, nose, or ears during food production (40%); had no protection from their long hair (32.2%) while working with food; had inadequate working clothes (30.5%); and the payment process was not properly separated from food production (28.7%). Based on the observations of the respondents, it can be said that the personal hygiene of street vendors was not fully compliant with hygiene standards.The respondents noticed irregularities in the hygiene conditions of food production as follows: overflowing waste bin in the production area (86.7% responses) and the presence of employees’ personal items  in the production area (44.7%).In the area of hygiene of food production and distribution processes, the respondents noticed that ready-to-eat products and wasted ones were not separately stored (79.2%), and they observed unauthorized people in the production areas (38.9%).Culinary preferences and eating habits have changed over time and are inextricably linked with human history. However, globalization is accelerating the pace of these changes more than in previous centuries. Economic progress, development of food techniques, and technology have brought many benefits, among them the speed of work and rest as well as the speed and convenience of obtaining and preparing a meal. On the other hand, living in a hurry, eating ‘ad hoc’ food or fast food, often non-compliant with the nutritionists’ recommendations, and diet-related diseases are a consequence of cultural change. Changes in eating habits occur not only between generations, but also among those who change their current model of nutrition to a new one in a relatively short time. The main causes of change in dietary preferences are factors such as migration, new ways of processing and storing food, international trade development, increases in levels of wealth, changes in family functioning, an increase in environmental awareness, fashion, etc. .Globalization causes the culinary traditions to be transferred from one place  to another. However, it can be a kind of threat, leading to the universalization of eating habits and the disappearance of local culinary traditions . Poland The interest in, and increased use of, catering services in Poland is due not only to having insufficient time to prepare meals, but also individuals lacking skills to prepare them, greater women’s involvement in professional work and returning home late with too many professional duties outside the home, an increase of 1–2 person households, as well as the increased income of Poles .Catering establishments take advantage of consumers’ interest in eating out and adjust their offerings to various social groups and their expectations and financial capacity. The constant rush and lack of time to eat and prepare a meal, as well as the relatively high cost of meals in traditional gastronomy, are the factors contributing to street gastronomy gaining popularity in Poland. Because it is mobile, it can reach consumers directly and offer simple dishes close to workplaces, tourist attractions, and other frequently visited places. All respondents participating in our research used street food services, 87% of which did so regularly.This is a result of the growing popularity of street food not only in Poland but all over Europe . In PolaIn the group of consumers assessed, the choice of burgers and kebabs dominated, having become very popular. This is one example of the transfer of eating habits from one culture to another. In Poland, street food, especially offered in food trucks, plays an important role in promoting the cuisine of other countries, often for exotic-craving Poles. The average Polish consumer, when dealing with street food dishes, has the opportunity to learn new tastes, which may have an impact on changing current culinary preferences.Owners of street food outlets develop their own gastronomic activity based on their impression of and fascination with other cultures and cuisines. For example, ‘Carnitas Food Truck’ (in Warsaw) specializes in street food typical for Mexican cuisine such as tacos, burritos with salsas chili habanero, chipotle, and guacamole. The food truck ‘La Chica Sandwicheria’ (in Warsaw) specializes in food typical of Cuban cuisine. Sometimes globalization of cuisine is seen in examples such as the food truck ‘Pepe Crepe’ (in Warsaw) that offered Japanese-style crepes that are not traditional but contemporary, globalized versions of crepes. Street food is offered at many culinary festivals in Poland, such as Slow Weekend and Asian Street Food Fest . As resen = 1013) visits foodservice outlets once a month, with the individuals being slightly more often men and far more often young people, and those with middle and higher income. People with secondary and higher education benefit more commonly from food services, including school and university students and white-collar workers, who live in large cities. It should be mentioned that participants in our study were mainly people under 30 years of age (about 85%), with secondary and higher education (91.5%), and who lived in a big city (about 70%).Respondents used street food usually two or three times a week and once a week. Our results are similar to the results of Kowalczuk , who hasBased on the analysis of clusters due to the choice of menu, preferences of street food consumers were identified in our study. They are: ‘burger-enthusiasts’, ‘kebab-enthusiasts’, ‘ice-cream enthusiasts’, and ‘non-specified-oriented consumers’. Levytska and Kwiatkowska  have indStreet food consumers mentioned the following main reasons for eating outside the home: getting to know new tastes, socializing, convenience and lack of time, celebrating special occasions, and reluctance to prepare dishes on their own. Other authors have also pointed out reasons such as convenience and saving time .The main reasons for consumers using catering outlets are the taste of dishes, reasonable prices, convenience, and saving time, while the main barrier is a lack of money. Consumers tend to spend more money in catering outlets per month than in the past , and thePolish respondents use a variety of sources when choosing a catering establishment, including street food outlets. Most often these are the opinions of friends and family, social networks and Internet forums, websites with restaurant reviews, articles on the Internet and articles in the press, blogs, and others. This is a typical behavior in the era of globalization. It has been observed that the following mass media sources play an important role in promoting catering services and influencing culinary tastes in Poland: daily press, weeklies and monthly magazines, television , and especially the Internet  . AccordiAmong the factors in our study determining the selection of catering establishments, the most important role was played by ‘preferences and quality’ and ‘economic and food safety’. The most important discouraging factors were ‘quality’ and ‘economic and hygiene factors’.According to market reports ,81, respn = 1000), pancakes (32%), casseroles with bread (31%), burgers , and Belgian fries .The form of street food in Poland is typical in developed countries . DespiteVarious authors have tried to characterize the most popular dishes found in mobile gastronomy in developing countries  and founThese results are similar to those of other authors. Different studies state that street food meals are characterized by low nutritional value and contain large amounts of carbohydrates and fats, especially saturated fats , and strMoreover, many studies ,90,91,92According to Nonato et al. , street Most respondents have a positive opinion of street food outlets. Consumers do not consider them to be a cheap way of eating meals and do not agree with the statement that these are premises with a low level of hygiene. They believe that such meals are as safe as those offered in stationary premises.Although the greatest problems and the greatest negligence related to ensuring food safety occur in developing countries, this does not mean that in Europe and the U.S. ,38,39,94In street food outlets, meals are prepared in the presence of consumers. It is worth disseminating the results of similar studies among a wide audience to increase food safety awareness among consumers. Consumers are able to enforce proper food safety and should react to non-compliance with hygiene rules. The situation reported in Ghana is not recommended . DespiteThis study has some limitations in terms of both its methodology and its applicability. The sample selected for the study consisted mainly of young adults between 19 and 30 years; therefore, caution should be exercised in attempting to generalize the results to an entire population. In addition, the study was conducted only in large cities in Poland. Consumers’ perceptions and behavior may be different in other places. Another limitation is that the consumers of street food were from only one country. Despite the limitations, the results obtained are of practical importance, especially for food safety authorities and street food vendors and owners.The forms of street food in Poland are forms typical in other developed countries, as evidenced by the identified consumer profiles. About half of the respondents were ‘kebab- and burger-enthusiasts’. Similar factors affect the behavior of Polish consumers regarding the choice of street food outlets and the lack of willingness to re-use this form of meal outside the home. Consumers’ preferences, pro-quality factors, such as quality of services, quality of meals, hygiene, and food safety, and economic factors, such as the price of a meal, are of primary importance.The opinion of the surveyed consumers about food street outlets was not always positive. Consumers commented on many aspects of sanitary conditions of street food facilities, such as personal hygiene, hygiene conditions of food production, as well as hygiene of food production and distribution. For these reasons, the presented research is of practical importance for managers of these facilities and official quality control.The results of the study can also be helpful for street food entrepreneurs because of the indicated identified promotion channels. It has been shown that the Internet is an important source of information on catering establishments, and social networking sites play a large role in consumer traffic and can be used for promotional purposes, practically without generating costs.The quality of dishes, the quality of service, and hygiene are the factors that make consumers use a catering establishment. Constantly increasing consumer awareness in terms of proper nutrition and proper hygiene of food production may contribute to the improvement of quality in street food outlets.Good sources that can encourage hygienic awareness by consumers come primarily from mass media, including the Internet and the website of sanitary inspections. Consumers should be made aware that proper food quality is both an appropriate nutritional value related to good manufacturing practice (GMP) and a result of food safety related to good hygiene practice (GHP). Consumer knowledge regarding what they should take into consideration when buying street food can be more effective than control and can better counteract improper hygiene practices of vendors.The solution could be to create a street food digital application in which consumers could find information about street food and its nutritional value, but also could share their opinions on offerings, including their quality and observed production hygiene.


Reversible splenial lesion syndrome (RESLES) is a single-stage non-specific syndrome with unclear pathogenesis. There has been no report on answer delay in patients with RESLES. We report a female patient who was admitted to our department for mixed aphasia accompanied by cognitive impairment. During the rapid improvement of aphasia, there was a clear phase of language output response delay accompanied by resolution of imaging lesions. We analyzed the course and the examination results of the patient and speculated the cause and pathogenesis. RESLES-relevant knowledge was systematically reviewed, which will help doctors in the classification of cerebral function and the diagnosis of RESLES. The specific language and cognitive impairment may be associated with the damage of contact fibers in the bilateral primary and secondary sensory and motor cortices. It is characterized by reversible round or oval, non-enhanced lesions in the corpus callosum on magnetic resonance imaging (MRI). According to the characteristics displayed on MRI, RESLES is classified as type I, only involving the splenium of the corpus callosum, and type II, involving the white matter and/or whole carcass . The pat22.1A 54-year-old female patient, junior high school education level, was admitted to the hospital with intermittent watery diarrhea for 7 days and progressive speech disorder for 3 days. The cranial computed tomography showed no abnormality at the community hospital and she was immediately referred to the hospital. There were no symptoms of numbness, fatigue, disturbance of consciousness, limb convulsions, urinary incontinence, and fever since the onset and no family history or previous history of neurological disorders. However, due to the swelling of the buttocks for nearly 20 days, the patient applied self-purchased drugs on bedsore wounds in the hip area, while receiving an intravenous infusion of penicillin.Informed consent: Informed consent has been obtained from the patient who participated in this study.Ethical approval: The research related to human use has been complied with all the relevant national regulations and institutional policies and performed in accordance with the tenets of the Declaration of Helsinki and has been approved by the authors’ institutional review board or equivalent committee.2.2On admission, she was conscious and silent, with a temperature of 36.8°C. The patient’s visual space and executive ability, attention, abstract thinking, and short-term memory were significantly impaired. Her answers to questions were delayed by 8 s, even after multiple repetitions. The Montreal Cognitive Assessment (MoCA) score was 3 points, and the normal value range is not less than 26 points.9/L, the ratio of neutrophils was 91.8%, and the proportion of lymphocytes was 4.5%. The patient was positive for herpes simplex virus IgG antibodies and a titer of 161.10 U/mL for cytomegalovirus IgG antibodies; serum potassium concentration was 3.2 mmol/L. Renal function, liver function, routine stool test, blood culture for bacterial infection, erythrocyte sedimentation rate, D-dimer, electrolytes, cerebrospinal fluid, and other related examination results were normal. The electroencephalogram showed an extensive and mild abnormality of the adult type, and the slow-wave band value was high.The patient’s right leg was disabled, with severe weakness and pathological reflex. The right upper limb had an increased tone, whereas other neurological examinations showed no abnormality. Leukocyte counts were 10.75 × 102.3boomerang sign. The genu of the corpus callosum also showed relatively high signal intensity on DWI  images and increased signal intensity on T2-weighted (T2W) images and fluid-attenuated inversion recovery (FLAIR). The splenium of the corpus callosum demonstrated relatively decreased signal intensity on T1W images and increased signal intensity on T2W images. Its signal intensity was slightly higher than that in normal brain tissue on FLAIR. Diffusion-weighted imaging (DWI) showed hyperintensity of it resembling y on DWI . Bilatery on DWI . On day y on DWI . Moreove−3 mm2/s .2.4According to the patient’s status at the time of admission, the initial diagnosis was central nervous system infection, and the patient was given antiviral and dehydration treatments during hospitalization. From the ninth day after the onset, the patient’s symptoms, including the mixed aphasia, rapidly improved, and the diagnosis was corrected based on the above-mentioned magnetic imaging features.Sixteen days after the onset, significant recovery was observed in all cognitive subdomains, she answered questions in 4 s, and the MoCA score was 7 points. Twenty-eight days after the onset, the cognitive subdomain impairments were restored to normal, she responded to questions in 1 s, and the MoCA score was 21 points.3In 1999, Kim et al. first reported reversible corpus callosum lesions . In 2004Although patients with RESLES have no specific clinical symptoms, answer delay is rare even in other diseases. Patients with RESLES often have prodromal symptoms, including fever and diarrhea, followed by seizures, confusion, ataxia, lethargy, headache, paralysis, etc. . RESLES RESLES should be differentiated from diseases such as acute cerebral infarction, acute disseminated encephalomyelitis, and extrapontine myelinolysis. The symptoms of the case that we report were self-limiting and imagings revealed a reversible lesion resembling the boomerang sign that distinguishes it from other diseases and should be classified as type II. Most of the earliest cases were patients with epilepsy or infants. Due to the widespread use of MRI, various cases associated with drug, malignancy, infections, subarachnoid hemorrhage, metabolic abnormalities, and trauma have been emerged . A few cN-methyl-d-aspartate receptors, sodium–potassium pumps, etc., which results in water flow into astrocytes and neurons [The pathogenesis of RESLES is not clear. Various pathogenic factors yield the same imaging features indicating that these pathological mechanisms must involve some common factors that should be explored. Starkey et al. proposed that the mechanism of cytotoxic lesions of the corpus callosum is the excitotoxic action of glutamate on  neurons . The den neurons . Other t neurons  and gene neurons .The corpus callosum, located at the bottom of the fissura longitudinalis cerebri, is a transverse nerve fiber bundle connecting the left and right hemispheres of the brain. It plays an important role in the transmission and integration of writing, body, face, and visual information. Therefore, our patient’s silent state on admission may be caused by the disconnection of the two hemispheres and the disruption of cerebral cortex functioning . After t


Several versions of the dot probe detection task are frequently used to assess maladaptive attentional processes associated with a broad range of psychopathology and health behavior, including eating behavior and weight. However, there are serious concerns about the reliability of the indices derived from the paradigm as measurement of attentional bias toward or away from salient stimuli. The present paper gives an overview of different attentional bias indices used in psychopathology research and scrutinizes three types of indices  calculated from a pictorial version of the dot probe task to assess food-related attentional biases in children and youngsters with and without obesity. Correlational analyses reveal that dynamic scores  are dependent on general response speed. Reliability estimates are low for the traditional and probability indices. The higher reliability for the dynamic indices is at least partially explained by general response speed. No significant group differences between youth with and without obesity are found, and correlations with weight are also non-significant. Taken together, results cast doubt on the applicability of this specific task for both experimental and individual differences research on food-related attentional biases in youth. However, researchers are encouraged to make and test adaptations to the procedure or computational algorithm in an effort to increase psychometric quality of the task and to report psychometric characteristics of their version of the task for their specific sample. Different theoretical accounts on problematic eating, overweight, and obesity propose that food stimuli automatically attract visual attention, particularly in individuals with overweight and weight concerns . 59.64% of the participants were recruited in the WELCOME-project  in this subsample were all obese . The remaining participants  were recruited in convenience samples by Master students at Ghent University under supervision of LV and TN. They were all normal weight . This sample size was justified by data availability: all data that were collected at Ghent University, using this particular dot probe procedure between 2017 and 2020 were used. As such, the sample size is sufficiently large to detect group differences of d = 0.4, which is considered the smallest effect size of interest in psychology , aged between 7 and 19  was calculated by dividing measured BMI (weight in kg/squared length in cm) by norm BMI for age and sex, and multiplying this by 100. Norm BMI for age and sex was determined as the 50th percentiles of the BMI for age and sex based normative data. An adjBMI equal to or smaller than 85% is considered underweight, equal to or >120% as overweight, equal to or >140% as moderate obesity, equal to or >160% as extreme obesity.www.food-pics.sbg.ac.at Blechert et al., Attentional bias toward food-related stimuli was measured using a pictorial version of the dot probe task  or incorrect responses were excluded.Traditional Attentional Bias Score (ABS)  , is taken. Again, higher values indicate stronger effects, either toward or away from food.with Evaluating the different indices was done stepwise, vis-à-vis the considerations below.RT-based effects, as the ABS and the TL-BS are known to be inflated for individuals responding slowly Fazio, . Since gSplit-half reliability of these indices  will be estimated by Pearson correlations between index scores calculated in both test halves, for the total group and for both weight status groups separately. To test if reliability was influenced by general response speed, linear models predicting performance in first test half by reliability in second test half and RT were computed.Based on the theoretical assumptions on problematic eating and overweight and obesity, a significant difference can be expected between youth with obesity and youth with normal weight on their reaction to food vs. neutral stimuli. This will be tested using linear mixed models  with weight status group as dependent variable (dummy coded), and the indices as independent variables will be reported. To control for general response speed, mean RT on neutral-neutral trials will also be added as between-subject variable. If an index would be a meaningful measurement of food-related attentional bias in experimental research for this youth sample, weight status would be significantly predicted by the index, with no significant effect of general response speed.Because attentional bias for food is thought to be stronger in individuals with higher weight, the linear association between the attention bias indices and adjusted BMI will be estimated by Pearson correlations. If an index would be a meaningful individual differences variable, significant positive correlations would emerge.n = 337), and both weight groups separately .B = −0.001, SE = 0.001, t(328) = −2.03, p = 0.04) none of the models predicting weight status by attention and general response speed reached significance , with multiple R-squared and adjusted R-squared ranging between R2 = 0.002 (for mean TL-BSpos) and R2 = 0.01 [for mean TL-BSneg and Abs(PI-0.5)], and between adjR2 = −0.004 (for mean TL-BSpos) and adjR2 = 0.008 [for mean TL-BSneg and Abs(PI-0.5)], respectively.The present study investigated whether attentional bias toward food could be meaningfully assessed in a youth sample of children with and without obesity, using a pictorial version of the dot probe task. The rationale for this study, was grounded in the widespread practice to use the dot probe procedure to measure and modify food-related attentional bias, both in experimental laboratory studies and in clinical intervention studies  showed higher and significant correlations, comparable to those reported by their developers , When using this test set up, administered to this specific sample , to compute these specific indices , we were unable to provide evidence for the task's applicability to assess food-related attentional biases. However, these conclusions pertain only to this test version, in this sample in this context  (Rodebaugh et al., The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Ghent University FPPW. Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin.LV and TN share project administration. LV and MD contributed to conception and design of the study. LV and MB did the literature search. TN collected data and wrote down the data management plan. MB organized and prepared the database. LV, MB, and MD performed the statistical analysis and wrote sections of the manuscript. LV wrote the first draft of the manuscript. All authors contributed to manuscript revision, read, and approved the submitted version.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.