Salariopsis fluviatilis from Italian geographic areas (north-western regions and Sardinia Island) so far never investigated and uses five species delimitation methods in order to shed further light on the Mediterranean phylogeography of this fish and give a more comprehensive scenario of its taxonomic status. Our findings clarified the phylogeographical patterns of the northern Italian populations of Salariopsis fluviatilis sensu stricto, considering the Po River basin and some Tyrrhenian basins of the Liguria region on the other side of the Ligurian Alps. The dispersal pathways of the island of Sardinia were also investigated suggesting that patterns of genetic structuring for this species are probably linked to Pleistocene glacial and interglacial periods. Results obtained supported previous studies in evidencing the occurrence of a species complex for Salariopsis fluviatilis with at least three taxonomic units: Salariopsis fluviatilis sensu stricto, Salariopsis cf. fluviatilis diffused in the Middle East and a further taxonomic entity from the Iberian Guadiana River basin whose tributaries flow in the Atlantic Ocean. For what concerns Salariopsis fluviatilis sensu stricto, two divergent groups of populations were reported in the present study, being the first diffused in western Mediterranean areas and the second in western Adriatic and eastern Mediterranean areas.The present study provides new molecular data on populations of the freshwater blenny Salariopsis (Blenniidae) comprises freshwater blenny fish that inhabits Mediterranean Sea, Black Sea, and north-east Atlantic areas. Three species were formally described to date: Salariopsis fluviatilis. S. economidisi, and S. atlantica. In this study, 103 individuals were collected from different Italian regions  and analyzed using the mtDNA Control Region and the ribosomal 16s gene. We aimed (i) to depict the phylogeographic patterns of S. fluviatilis in northern Italy and Sardinia and (ii) to compare the genetic structure of Italian samples with those from other Mediterranean regions. Results obtained showed the presence of a well-supported genetic structuring among Italian S. fluviatilis populations, shedding new light on the phylogeographic patterns of northern Italian populations of S. fluviatilis sensu stricto across the Ligurian Alpine ridge and the Sardinia Island-mainland dispersal patterns. Furthermore, our species delimitation analysis was consistent in supporting results of previous research about the presence of genetic differentiation among S. fluviatilis, evidencing: (i) a large group of S. fluviatilis sensu stricto that includes two sub-groups , (ii) one group comprising populations from the Middle East of a taxonomic entity corresponding to Salariopsis cf. fluviatilis, and (iii) one group of Iberian individuals from the Guadiana River.The genus  Salariopsis fluviatilis  (Actinopterygii: Blenniiformes: Blenniidae), is a small benthic fish that inhabits river basins and lakes around the Mediterranean Sea. It belongs to the recently described genus Salariopsis Vecchioni, Ching, Marrone, Arculeo, Hundt & Simons, 2022, proposed by Vecchioni and co-authors [Salaria Forsskål, 1775. Nowadays, the genus Salariopsis includes freshwater species whose distribution is across the eastern Atlantic coasts (Morocco), the Mediterranean Sea, and the Black Sea [The freshwater blenny, is Asso, 801 , endemic to Lake Trichonis in Greece [Salariopsis atlantica Doadrio, Perea & Yahyaoui, 2011, endemic to the Seboui river basin in Morocco [In this context, in addition to  Morocco .S. fluviatilis shows, within the genus, the widest range of distribution, its populations are generally small and highly localized [Salariopsis species, which show confined  or even point-like  distributions.Although ocalized ,21,22,23S. fluviatilis is highly susceptible to several anthropomorphic disturbances and to global environmental change effects. Nevertheless, even though it has been considered vulnerable or endangered in several countries and reported in Appendix III of the Bern Convention, S. fluviatilis has been evaluated by the International Union for Conservation of Nature (IUCN) as data deficient (IUCN 2013), due to a lack of data on its abundance and population trend.Similar to other freshwater fish living in fragmented habitats, S. fluviatilis [Salariopsis in the Mediterranean area, with inferences reported also for S. fluviatilis. Wagner et al. [S. fluviatilis populations: (1) Middle East ; (2) Guadiana (Guadiana basin with the Zùjar and Esteras rivers that flow on the European Atlantic coast); (3) Algeria-Verde (in southern Spain and Algeria with the Verde River and Oued Boughzazene); (4) Occidental basin , (5) North Oriental basin , and (6) the Island of Crete (Greece). The results of Wagner et al. [Salariopsis fluviatilis. Remarkably, based on the results of two methods of species delimitation, Belaiba et al. [S. fluviatilis from the Middle East Mediterranean region should be regarded as a differentiated species, i.e., Salaria cf. fluviatilis. However, in light of the description of the new genus Salariopsis [Salariopsis cf. fluviatilis hereafter.Despite the potential interest of this species from phylogeographic, taxonomic and conservation perspectives, not many studies have been performed to depict the genetic structure and phylogeography of viatilis ,20,24,25r et al. , in partr et al.  also supr et al. ), suggesa et al.  suggest lations:  Middle ES. fluviatilis in the western Mediterranean [S. fluviatilis in the Mediterranean would be the product of the recent spreading of individuals from the two glacial refuges of the Iberian Peninsula and Corsica Island. However, in this study, the lack of samples from Liguria  probably affected the possibility to fully reconstruct the dynamics of colonization which involved Corsica Island. Indeed, Wagner et al. [S. fluviatilis from Corsica, highlighted the importance of focusing studies on island populations due to the peculiarity and fragility of the island’s fish fauna, to infer the biogeographical dynamics of the populations that live there. These same authors also expressed hope for extending the geographical sampling to provide a more comprehensive scenario of S. fluviatilis in the Mediterranean area.Furthermore, two main studies focusing specifically on the nuclear genetic variation of erranean ,25 suggeerranean . In partr et al. , analyziS. fluviatilis is distributed in a discontinuous way, starting from the northern regions and continuing through the Tyrrhenian side, down to Campania, Sardinia, and the Sicily islands. Isolated populations have also been found in Calabria and on the Adriatic coasts of the peninsula [S. fluviatilis and the country is characterized by different (hydro)geographic boundaries that strongly shaped its ichthyofauna with the presence of several endemic species—64 taxa are considered native to inland waters of Italy [S. fluviatilis living on Sardinia Island deserve particular attention, as this species is one of the few putative native freshwater fish known for the island [S. fluviatilis from Italian areas that are not yet under investigation. Indeed, knowledge of the genetic structure of populations from north-west Italy and the Sardinian Island could help to fill in some gaps in the mosaic of S. fluviatilis genetic distribution in the central-western Mediterranean.Focusing on Italy, eninsula ,27. Althof Italy  and amonof Italy —no wide-e island . The fole island ,32,33,34S. fluviatilis populations, sampled in Sardinia and north-west Italy  in order to shed light on the phylogeography of the populations now living in the main central-western Mediterranean islands and the north-west Italian mainland; (2) to compare the genetic structure of our samples with that of other Mediterranean regions to put them in a wider geographic framework; and (3) to make an attempt to further clarify the taxonomy of the species across its whole range of distribution by using different methods of species delimitation to evidence, if any, distinct evolutionary lineages within S. fluviatilis that deserve to be deeply investigated in the future.As a results, the goals of our paper were: (1) to depict the phylogeographic patterns of S. fluviatilis allowed us to compare the sequences obtained in the present study with many others from different geographic areas. In previous studies, the combined use of Control Region and 16s sequences has been demonstrated to be helpful in providing insights into the genetic structuring and phylogenetic history of S. fluviatilis populations [Two portions of the mitochondrial Control Region and of the 16s ribosomal gene were used as markers. The use of the mtDNA, which is maternally inherited, is considered an appropriate choice to solve intraspecific taxonomic and phylogenetic issues and to depict evolutionary lineages ,36,37. Fulations ,15,17.Salariopsis fluviatilis were caught using an electric stunner from the freshwaters of 4 different Italian regions  between February 2019 and March 2022  and immediately transferred to a recovery tank before being released. Tissues were preserved in absolute ethanol and used to perform DNA extraction. The above reported sampling method was approved by the ethics committee of the University of Sassari (Prot n. 122 770 of 7 November 2022), and its researchers led the sample collection activities during the present study.The protocol of sampling and analysis of the fish fauna of wadable lotic systems, provided by the Italian Higher Institute for Environmental Protection and Research (ISPRA) , was folSardinian samples were collected in three tributaries of the Flumendosa river , all confluent in the Alto Flumendosa man-made reservoir (originated in the middle of the last century by a dam) , the Rio Mannu di Scano Montiferro river , and the Rio Mannu di Posada river (north-east Sardinia); Ligurian samples were collected in the Entella river (east Liguria) and in the Roja river (west Liguria); samples from Piedmont were collected in the San Giovanni river, in the San Bernardino river, and in the Strona di Omegna river; the sample from Lombardy was collected in the Tartaro Fuga creek, tributary of the Oglio river.2 was set at 200 µM and 1.5 mM, respectively. PCRs were performed in a GeneAmp PCR System 9700 Thermal Cycler , programmed as follows: 1 cycle of 4 min at 94 °C, 35 cycles of 30 s at 94 °C, 30 s at 51 °C (for both Control Region and 16s gene), and 30 s at 72 °C. At the end, a post-treatment of 10 min at 72 °C and a final cooling at 4 °C were carried out. Both positive  and negative controls were used to test the effectiveness of the PCR protocols and the absence of possible contaminations. Electrophoresis was carried out on 2% agarose gels, prepared using 1× TAE buffer  and stained with Gel Red Nucleic Acid Stain . PCR products were purified by ExoSAP-IT  and sequenced for forward and reverse strands (by means of the same primers used for PCR), using an external sequencing core service .Total genomic DNA was isolated from a portion of fin tissue using the Macherey-Nagel Nucleo Spin Tissue Kit , following the supplier’s instructions. DNA solutions were quantified using the Nanodrop™ Lite Spectrophotometer , which showed an average yield of 29 ng/µL. A portion of the mitochondrial Control Region and rRNA gene 16s were amplified by standard PCR using the following primers: CR-F (forward) (5′-CCACTAGCTCCCAAAGCTA-3′) and CR-R (reverse) (5′-CAGGACCAAGCTTTTGTGC-3′) ; 16s Forhttps://www.ebi.ac.uk/Tools/msa/clustalo/ (accessed on 16 September 2022)) and deposited in GenBank (OP675744-OP675846 for the Control Region and OP653611-OP653713 for 16s).A total of 103 newly generated sequences was aligned using the package Clustal Omega   available on GenBank to date (last update 30 September 2022) (see The Control Region and 16s 022) see .S. economidisi  and S. atlantica . Furthermore, the marine species Salaria basilisca (MW555061) and Salaria pavo (MW555062) were used as outgroups. As well, in the 16s dataset, the sequences available for the Mediterranean freshwater blenny species were also included, i.e., S. economidisi  and S. atlantica . The marine species—S. basilisca (MH724820) and S. pavo (FJ465707)—were used as outgroups , number of haplotypes (H), haplotype diversity (h), and nucleotide diversity (π) using the software package DnaSP 6.12.03  .To test the reliability of the three datasets  for taxonomic and phylogenetic analyses, the phylogenetic signal was checked by the software TREEPUZZLE  : the likThe best probabilistic model of sequence evolution was achieved using jModelTest 2.1.3  , with a www.fluxus-engineering.com) (accessed on 12 October 2022) . Transitions and transversions were equally weighted. Due to the absence of information about the possible appearance of retro-mutation events, the same weight (10) was assigned to each observed polymorphism.To infer the genetic relationships among haplotypes and to detect the possible occurrence of discrete genetic clusters, a median-joining network  was consSalaria from GenBank: they grouped 280 sequences for the Control Region (see http://tree.bio.ed.ac.uk/software/figtree/) (accessed on 18 September 2022) .S. fluviatilis sequence was checked using five different methods of species delimitation. The GMYC  model tests for a significant shift in the branching rate along an ultrametric species tree. The analysis was performed on the ultrametric species tree obtained from the Bayesian dating analyses by means of the SPLITS (SPecies LImits by Threshold Statistics) package [http://r-forge.rproject.org/projects/splits/)(accessed on 18 September 2022). Species entities were identified by means of the single threshold option, which uses a single threshold in order to specify the transition from between- to within-species branching .http://species.h-its.org/ptp/) (accessed on 18 September 2022) on the phylogenetic species trees using default options and 500,000 MCMC generations. To test the reliability of results, each run was examined for convergence by visualizing the likelihood plot: if convergence occurred, the chain should stay at high likelihood locations during the run  ) uses tha et al. ).http://wwwabi.snv.jussieu.fr/public/abgd/abgdweb.html) (accessed on 18 September 2022) using the default settings. The correct species estimate was selected, according to Puillandre et al. [p = 0.001   method ie et al. , using ta et al. ).https://cran.rproject.org/) (accessed on 18 September 2022). The script partitions taxa into entities applying the fixed threshold of 2% obtained by Hebert et al. [The NDT (Nucleotide Divergence Threshold) method was used on each gene separately, by means of a script developed by Scarpa et al. , writtent et al. , using at et al.  genetic t et al. ).The principal coordinates analysis (PCoA) was performed using GenAlEX 6.5  on a matTripterygium delaisi, in a previous study by Koblmüller et al. [The software package Beast 1.10.4  was usedr et al. . Specifir et al. . Site par et al. . The sofr et al. . Tree Anr et al. .A total of 103 sequences for both Control Region (302 bp) and 16s gene (547 bp) were obtained in the present study. Both datasets were merged to Genbank sequences (280 for the Control Region and 136 the 16s gene) and used to implement phylogeographic analyses.Salariopsis fluviatilis  through Basic Local Alignment Search Tool (BLAST) analysis implemented in the GenBank nucleotide database (www.ncbi.nlm.nih.gov) (accessed on 24 August 2022) that showed a percentage of identity ranging from 97% to 100% for the Control Region and from 90% to 100% for the 16s gene.All the newly generated sequences were identified as belonging to the species S. fluviatilis . For the two samples from Lombardy only one haplotype was retrieved, while higher levels of divergence were found for samples from Liguria and Piedmont with 8 and 4 haplotypes, respectively.Regarding the 103 sequences obtained in the present study for the 16s gene, low levels of divergence were found for each region with a total of 7 polymorphic sites resulting in 8 haplotypes see . SamplesWith the aim to obtain a dataset including the level of polymorphism and informativeness as high as possible, these two datasets of sequences (Control Region and 16s) were concatenated to obtain a longer mitochondrial fragment, which was 849 bp long, to perform phylogeographic analyses. In this concatenated dataset, 23 polymorphic sites were found, resulting in 20 haplotypes and moderately low levels of haplotype and nucleotide diversity see . A similS. fluviatilis in Italy, the two Italian sequences (from the Lombard side of Lake Garda and Sicily) which had been previous published [The network analysis was performed on a concatenated (Control Region and 16s) dataset which included samples from northern Italy and Sardinia see  from theublished  were incGroup A included individuals from Sardinia and Liguria with the occurrence of two main star-like shapes involving the two areas. In particular, among samples from Liguria, a high level of variability was witnessed by the number of different haplotypes that are present in this area with a strong founder effect, as suggested by the occurrence of a very common central haplotype , which is surrounded by several derived haplotypes diverging for a few mutations. The haplotypes that likely originated from the lineages most common in Liguria, are private to single Ligurian individuals  or shared among Sardinian and Ligurian individuals. In only two cases, haplotypes likely derived from the Ligurian ancestor lineage are exclusive to Sardinian individuals. In particular, one of these two latter cases corresponds to the most common haplotype among Sardinian individuals, which was found in 46.8% of specimens collected and is private to Sardinia. In particular, only one haplotype isolated in the north-east of the island (Rio Mannu di Lodè river), which is exclusive to a single individual, could have not necessarily originated from the most common Ligurian and Sardinian founder lineages.Group B included samples of Piedmont and Lombardy from the present study with a marked founder effect suggested by the occurrence of a star-like shape. The two identical sequences found in Lombardy correspond to the most common haplotype occurring in Piedmont. Two out of three derived haplotypes were private of single Piedmontese individuals. The sequence from Lake Garda (Lombardy) from a previous study  was inclx-axis and y-axis and accounted for only 13.49%.In the PCoA graph see  the 61.3x-axis between two main groups (Groups A and B). Group A included individuals from Liguria and Sardinia with an internal, although poorly supported, sub-structuring. Indeed, an eastern Sardinian sample from the Rio Mannu di Lodè river .Cluster B was representative of northern Italy  and Sicily, along with the eastern part of continental Europe . Furthermore, three sequences from Spain and one from Portugal from the basin of the Guadiana River, which flows into Atlantic Ocean, set 6 to 9 point mutations apart from clusters A and B. Interestingly, a group of two Spanish and two Algerian sequences, all collected throughout a geographic area facing the Alboran Sea, sets in the network on a separate position between clusters A and B diverging for 4 point-mutations from each of them.S. cf. fluviatilis, along with sequences from Syria and Israel. Within this cluster, all the haplotypes found in Israel were exclusive to this area, while a common haplotype was shared between Turkey and Syria. Cluster C diverged from the previous reported Guadiana River group for 22 to 29 point mutations.Cluster C grouped the sequences from southern Turkey  that Belaiba et al.  identifiS. economidisi, endemic of Lake Trichonis in Greece, sets close to cluster B, 13 point-mutations apart. The two sequences representing S. atlantica, which was described in Morocco, were found 44 mutational steps far from cluster C, 50 from the Guadiana River group, and approximately 60 from clusters A and B.The sequences of S. fluviatilis together with the Mediterranean sequences taken from GenBank, and it split into a dichotomy characterized by two main clusters (clusters 1 and 2). The first cluster (cluster 1) showed a polytomic clade, including almost all S. fluviatilis sequences, and its sister clade representative of S. economidisi (represented by two sequences).The phylogenetic tree see  showed tS. fluviatilis grouped all the Italian sequences along with those from eastern Europe, mainland France, Corsica, Spain, Portugal, Switzerland, and Algeria. It includes some internal well-supported sub-clusters; among them, the three most relevant groups: (i) three Spanish and one Portuguese sequences corresponding to the Guadiana River group evidenced by network analysis, (ii) two Spanish and two Algerian sequences, and (iii) seven sequences from Crete. These three well-supported internal subgroups were also found by network analysis. The second cluster (cluster 2) of the tree included the Turkish, Syrian, and Israeli sequences, which grouped together in the cluster C of network analysis. Accordingly, with network analysis, an internal sub-structuring, consistent with the geographic distribution of samples, was also found in cluster 2 of the phylogenetic tree.The large polytomy of S. fluviatilis/S. cf. fluviatilis, the NDT and the ABGD methods converged in recognizing the same three entities. The first entity comprises Italian, French, Swiss, Algerian, Balkan and almost all Spanish sequences. The second entity includes the Turkish, Syrian and Israeli sequences corresponding to the group (including S. cf. fluviatilis) already retrieved by previous analyses. The third includes the Portuguese and Spanish sequences from the Guadiana River basin .Salariopsis included in the datasets.On the other hand, the PTP method detects four entities, partially converging with the NDT and ABGD but grouping the Guadiana River entity within the “European” entity, and splitting the group of Turkish, Syrian, and Israeli sequences into three separate entities. The GMYC methods found a unique entity for all the sequences belonging to the genus x-axis while y-axis accounted for 18.47%.In the PCoA analysis , the 41.S. fluviatilis Mediterranean populations evidenced by previous analyses, distinguishing four groups: (a) Group 1, which includes sequences from Greece, northern Italy (apart from Liguria), Sicily, Croatia, Albania, and some sequences from the western coasts of Turkey facing the Aegean and the Marmara seas; (b) Group 2, which includes sequences from Sardinia, Liguria, Spain, mainland France, Corsica, Switzerland, and Algeria; (c) Group 3, which includes the Guadiana River sequences from Spain and Portugal; and (d) Group 4, which includes sequences from Turkey that Belaiba et al. [S. cf. fluviatilis, Israel and Syria. It is important to note that group 1 and group 2 diverged only on the x-axis which accounts for only 18.47% of variation suggesting that, according to Network analysis, these two groups are structured among each other but with a weak genetic divergence.Results are consistent with the differences among a et al.  identifiSalariopsis economidisi and S. atlantica, and the two marine outgroups, S. pavo and S. basilisca, set in the plot apart from the four S. fluviatilis/S. cf. fluviatilis groups.Salariopsis species dates back to 990 kya. The common ancestor of the group of sequences, including S. fluviatilis, S. cf. fluviatilis and S. economidisi (see SEGR2_FJ465541_1 and SEGR1_FJ465540_1 in the tree) dates back to approximately 800 kya. Salariopsis atlantica (see SAMA2_FJ465526 and SAMA1_FJ465527 in the tree), which originated in 390 kya, sets in the phylogenetic tree as an external clade to the cluster including S. fluviatilis, S. cf. fluviatilis and S. economidisi.The time calibrated tree , obtaineS. fluviatilis, which includes Italian, Eastern European, French, Corsican, Spanish, Portuguese, Swiss, and Algerian individuals, along with the sequences belonging S. economidisi, differentiated approximately 610 kya. Within this group, the common ancestor to S. fluviatilis diverged approximately 575 kya, while the common ancestor of S. economidisi dates back to approximately 130 kya. Within the large clade of S. fluviatilis, it was possible to obtain a molecular dating for the three well-supported groups of sequences reported in the previous analyses. One group corresponds to the sequences from the Guadiana River basin and dates back to approximately 180 kya , another group of Spanish and Algerian sequences from the Alboran Sea area, dates back to approximately 150 kya , and the last group, including Cretan sequences, dates back to approximately 210 kya .The large group of sequences of S. cf. fluviatilis sequences, date back to approximately 630 kya.The group of sequences, including the Turkish, Syrian and Israeli The low level of genetic variation found for the 16s dataset see  may haveS. fluviatilis and two further small clusters of sequences for (1) the Guadiana River basin and (2) western Turkey (S. cf. fluviatilis) with Israel, respectively. Cluster A includes only sequences from the Italian regions of Piedmont, Lombardy, Liguria, and Sardinia, while cluster B includes sequences from Italy (only from Lake Garda in the north of the peninsula and Sicily Island), Croatia, Spain, Greece, and Turkey (in the West of the country). All the 16s sequences obtained in the present study were included within cluster A.However, even taking into account the low level of polymorphism and the reduced number of available sequences for the 16s dataset, the network analysis  was consSalariopsis fluviatilis: phylogeographic and phylogenetic analyses were performed including a high number of Italian specimens from the north-west regions of the peninsula  and the Mediterranean island of Sardinia, which represents an area so far neglected by molecular studies focused on this species. Moreover, molecular dating based on the present dataset, which covers most of the range of distribution of this species, gave us the possibility to put in an updated time range of the evolutionary patterns previously described for the new genus, Salariopsis [The present study represents the first insight into the genetic variability of the Italian populations of ariopsis ,16. Finaariopsis ,16.S. fluviatilis populations and to infer the taxonomic status of this species. In particular, the use of the mitochondrial Control Region has been effective in identifying genetically divergent groups within the species.The mitochondrial DNA showed to be a suitable tool to illustrate phylogenetic and phylogeographic relationships among S. fluviatilis populations. Indeed, the group that includes Ligurian and Sardinian samples is strongly divergent from the group formed by samples from Piedmont and individuals from Lombardy and Sicily. The genetic structuring between S. fluviatilis from Liguria and Piedmont, which do not share common haplotypes among each other, may be ascribed to the geographic isolation between these flanking geographical areas. Indeed, the genetic pattern observed here can be explained considering that the presence in the Ligurian region of boundaries, represented by the Alps ridge in continuity with the Apennines, hindered the Ligurian individuals’ colonization of the northernmost Italian regions. This scenario is known for other species of Italian freshwater fishes as well, for instance, within the genus Telestes, Stefani et al. [Squalius, the same barrier secretes S. squalus in the basins afferent to the Po River drainage network and S. lucumonis in the Tyrrhenian [Emys orbicularis) (Testudines: Emydidae) [Unio spp.  [Our research evidenced a well-supported genetic structuring among Italian i et al.  reportedrrhenian ,70,71,72mydidae)  and alsoionidae) .S. fluviatilis from Piedmont and Lombardy. This is an interesting finding, as it does not support the role ascribed to S. fluviatilis as a dispersal vector for the glochidia of the mussel of the genus Unio from Sicily to Sardinia or vice versa, suggested by Marrone and co-authors [S. fluviatilis, a higher number of individuals, from different sites in Sicily, should be analyzed. A wider sampling campaign would also allow for enlarging knowledge on the genetic distribution of this island populations.Worth noting, the sole Sicilian individual present in our dataset, from Belaiba et al. , grouped-authors . At the -authors ,76,77,78S. fluviatilis from Piedmont and Lombardy can be the result of movements of individuals from the Balkans through the Po river paleo-drainage that during the last glacial maximum included some Balkan tributaries. Indeed, the Balkan Peninsula was one of the main glacial refugia during the Pleistocene period, probably acting as a crossroad of different developmental processes [The genetic relationship that was found in the present study between samples of rocesses . FurtherS. fluviatilis samples analyzed here has been found in populations from Sardinia. As expected, this finding reflects the typical evolutionary model of species dispersal on an island. Taking as a model the migration patterns reported for the Salmo trutta lineages [S. fluviatilis may be linked to the glaciation episodes that occurred during the Quaternary. Indeed, the climatic oscillations throughout this geological event may have promoted the migration of some individuals of S. fluviatilis  from anas Plaut  demonstrS. fluviatilis during the late Pleistocene) and the Sardinian islands were renewed [Syngnathus abaster [S. fluviatilis and those of the islands [Due to low sea levels during Quaternary climatic oscillations, Sardinia may have maintained contact with the mainland via the Island of Elba, and during the Last Glacial Maximum (21 kya), connections between Corsica  genetic differentiation between two main groups of S. fluviatilis populations within the Mediterranean basin; (2) a group of populations from the Middle East represented by individuals from southern Turkey , Israel and Syria which corresponds to S. cf. fluviatilis in accordance with Belaiba et al. [Wagner et al.  and Belaa et al. ; and (3)S. fluviatilis, sequences from Italy , Greece, Croatia, Albania, and part of Turkey (western coasts areas on the Aegean and Marmara seas) are representative of the North Oriental genetic cluster, whereas sequences from Spain, Italy (with Liguria and the Sardinia Island), Algeria, France mainland, and the Corsica Island represent the Occidental genetic cluster. Interestingly, individuals from the basin of the Guadiana River, which is the only investigated freshwater basin whose tributaries flow in the Atlantic, are slightly differentiated by the Occidental genetic cluster, suggesting an incipient genetic divergence of this population from the Mediterranean clade of S. fluviatilis.Within the Mediterranean clade of S. fluviatilis too [S. fluviatilis colonized the upper Guadiana basin starting from river catches of the Júcar basin, as likely happened to the strictly freshwater fish Luciobarbus guiraonis (Cypriniformes: Cyprinidae: Barbinae) [S. fluviatilis, at least for some months [The tolerance of freshwater blennies, and ilis too , to a wiilis too . In partilis too , Almada ilis too  and Lapoilis too , it coularbinae) ,88. Nonee months .S. fluviatilis, our results also evidenced the genetic divergence of the group of a few Spanish and Algerian sequences from the area of the Alboran Sea, and the group of sequences from Crete Island, which likely originated in the interglacial period between the Mindel and the Würm glaciations.Furthermore, in accordance with Wagner et al. , within S. fluviatilis population of Crete may be regarded as a peripheral isolate , namelyS. fluviatilis, which live at the edge of the distribution of the species. The Alboran Sea can be considered a geographic area where surface marine circulation patterns may directly shape the genetic variation of marine organisms, and indirectly act on the genetic variation of brackish and freshwater species, thus producing a genetic divergence with other Mediterranean populations .S. fluviatilis sensu stricto, the geographic isolation among populations and the reduced gene flow prompted the genetic divergence among Occidental and Oriental groups of populations. In such a context, natural fragmentation or secondary contact between populations that were separated in the past but are currently present in adjacent areas may have further helped to generate the genetic differentiation observed between Liguria and Piedmont in northern Italy. However, such a divergence is not enough to be completely retrieved by species delimitation methods as a trace of incipient speciation, but it is suggestive of a relevant trend of ongoing genetic divergence.Interestingly, within the taxonomic entity Salariopsis originated approximately 1 million of years ago, before the beginning of Pleistocene glaciations. Then, repeated glacial fluctuations may have contributed to shaping the genetic divergence among fragmented populations of freshwater blennies, which were substantiated by speciation events in the genus Salariopsis. In such a context, the current distribution of S. fluviatilis and S. cf. fluviatilis lineages is the consequence of the Pleistocene post-glacial events that occurred between the end of Günz and the beginning of Mindel glaciations and that copious events of deglaciation may have led the structure of S. fluviatilis populations to be further shaped by the passage through different levels of salinity across river networks [Based on the divergence times estimated for the genetic clusters retrieved in the present study, it is possibly suggesting that the common ancestor to the Mediterranean species of networks  and Pleinetworks .S. fluviatilis to develop into a species complex as a consequence of past geological phenomena and present geographical boundaries. In this context, it is interesting to note that S. fluviatilis is separated from S. cf. fluviatilis by a number of mutations (23) that is greater than the number of mutational steps that separates it from the species S. economidisi (15), which is endemic to the western Greek Lake Trichonis [S. economidisi originated at the end of the interglacial period between the Mindel and Würm glaciations being likely contemporaneous with the Guadiana River taxonomic unit and to Alboran Sea and Crete Island genetic clusters.It is reasonable to consider the possibility that the evolutionary stages evidenced in the present study led utations 3 that isSalariopsis fluviatilis, S. economidisi and the Guadiana River taxonomic unit share a unique common ancestor that likely originated during the Günz glaciation and that is contemporaneous with the ancestor of S. cf. fluviatilis. In accordance with Perdices et al. [s et al.  and Neats et al. , these tS. cf. fluviatilis taxonomic entity could be representative of a new species, distinct from S. fluviatilis, as S. economidisi and S. atlantica, which is pending a formal morphological description and new genetic analyses. In this context, it is interesting to note that S. cf. fluviatilis shows a genetic similarity higher with the species S. atlantica, which is endemic to the Seboui river basin in Morocco [S. fluviatilis sensu strictu. This finding suggests that the Mediterranean area of North Africa could be inhabited by different freshwater species belonging to the genus Salariopsis, which may be characterized by relevant levels of genetic structuring so far undetected. What is reported here is in accordance with the phylogeographical patterns observed in other Maghrebian freshwater species , which has promoted genetic exchanges among river basins.Interestingly, the genetic divergence found for the Guadiana River taxonomic entity and also for the two divergent clusters, found within S. fluviatilis and S. cf. fluviatilis (approximately 600 kya each) and of (2) the clusters of S. economidisi, Guadiana River, Alboran Sea, and Crete Island (on average 135 kya) suggest the contemporaneous origin of these taxonomic entities. This finding supports the occurrence of similar evolutionary rates for these groups produced by the perfect timing of the mitochondrial DNA molecular clock for all Mediterranean freshwater blennies whose common ancestors likely underwent the same selective pressure prompted by similar geological conditions and geographic barriers. On the other hand, our results point out that the origin of S. atlantica is more recent in respect of the differentiation of S. fluviatilis and S. cf. fluviatilis, dating back to the interglacial period (approximately 390 kya) between the end of Mindel and the beginning of Riss glaciations.Overall, in the present study, the similar temporal estimates obtained by molecular dating for the origin of (1) Salariopsis fluviatilis sensu stricto across the Ligurian Alpine ridge and the Sardinia Island-mainland dispersal patterns, with an origin and evolutionary trajectory of founders. In particular, this research contributes to put another piece in the mosaic of the molecular data of the Italian populations and helped to understand the processes of genetic differentiation involving Mediterranean island populations whose limited effective size and high degree of fragmentation make them ideal reference models for evolutionary processes.In the present study, the hypervariable mitochondrial Control Region was shown to be highly informative in depicting the phylogeographic patterns of Italian populations of Salariopsis. Indeed, our results support the possible occurrence of a complex of species for S. fluviatilis, namely S. cf. fluviatilis  in the a et al. ,37. Indeviatilis ,10,20,25viatilis .


General practitioners (GPs) are frequently patients' first point of contact with the healthcare system and play an important role in identifying, managing and monitoring cases. This study investigated the experiences of GPs from seven different countries in the early phases of the COVID-19 pandemic.International cross-sectional online survey.General practitioners from Australia, Austria, Germany, Hungary, Italy, Slovenia and Switzerland.Overall, 1,642 GPs completed the survey.We focused on how well-prepared GPs were, their self-confidence and concerns, efforts to control the spread of the disease, patient contacts, information flow, testing procedures and protection of staff.GPs gave high ratings to their self-confidence  and their efforts to control the spread of the disease . A decrease in the number of patient contacts , the perception of risk (5.3 95% CI 4.9–5.6), the provision of information to GPs , their testing of suspected cases  and their preparedness to face a pandemic  were rated as moderate. GPs gave low ratings to their ability to protect staff (2.2 95% CI 1.9–2.4). Differences were identified in all dimensions except protection of staff, which was consistently low in all surveyed GPs and countries.Although GPs in the different countries were confronted with the same pandemic, its impact on specific aspects differed. This partly reflected differences in health care systems and experience of recent pandemics. However, it also showed that the development of structured care plans in case of future infectious diseases requires the early involvement of primary care representatives. We focuInternational is part of the COVI-Prim project, which is described in detail elsewhere  self-confidence, (2) efforts to control the spread of the disease, (3) decrease in number of patient contacts, (4) perception of risk, (5) provision of information to GP, (6) testing suspected cases, (7) preparedness for a pandemic, (8) protection of staff; factor scores ranged from 0 to 10]. The questionnaire was transferred to LimeSurvey®  or SurveyMonkey® . Invitations to GPs to respond to the questionnaire were sent out based on the mailing lists of participating universities and local GP associations. As the lists probably overlapped, it is not possible to know precisely how many GPs were contacted and hence to calculate a response rate. Participants were first informed about the length of the survey, the investigators, and the purpose of the study. After the survey had been completed, all data on the online platform were stored in SPSS files. GPs received no financial incentive to participate. Surveys started at different time points in the participating countries. In most countries, GPs first began to answer the survey when the number of patients testing positively had begun to decrease. Completion rates ranged from 63.3% in Slovenia to 91.7% in Australia. The median time required to answer the questionnaire ranged from 11.0 min in Australia (interquartile range: 7.6–15.1) to 17.3 (IQR: 12.0–22.5) in Italy. Further details on completion rates and the time required to answer the questionnaire are reported elsewhere , as appropriate. Categorical variables are provided as absolute numbers and percentages. In the main analysis, environmental variables  that may have influenced the responses, and the role of the GP within the practice (employed vs. owner), were analyzed using General Linear Models. The main effects and all two-way interactions were analyzed. Bonferroni correction was used to take account of multiple testing. Estimated means and 95% confidence intervals were used to present the results. For a better understanding of the results, responses to the items on each scale were also presented. In this presentation, the response categories “yes” and “yes, probably” and the response categories “no, probably not” and “no” were combined. No statistical correction was carried out to adjust for non-representative samples.The study protocol was approved by the Ethics Committee of Bond University, Australia (AS200424), Goethe University Frankfurt, Germany (ID 20-619). The study required no ethical approval under Austrian, Italian, Slovenian, Swiss, and Hungarian law.n = 120), Austria (n = 434), Germany (n = 583), Hungary (n = 190), Italy (n = 112), Slovenia (n = 86), and Switzerland (n = 117). Mean age of the GPs was 51.9 years (SD: 10.8). The majority of GPs were male (52.1%) and practiced in a city with fewer than 20,000 inhabitants (56.6%). All demographic characteristics are provided in The survey was filled out by 1,642 GPs from Australia  and their efforts to control the spread of the disease . The decrease in the number of patient contacts , GPs' perception of risk (5.3 95% CI 4.9–5.6), the provision of information to GPs , testing of suspected cases  and their preparedness for a pandemic  were rated as moderate. GPs gave low ratings to their efforts to protect staff (2.2 95% CI 1.9–2.4).p = 0.043) and Italian GPs . More Austrian GPs were further convinced they knew how to provide the best possible care to their patients during the pandemic   significantly higher than Hungarian  .p = 0.003). The most pronounced differences between Slovenian and Hungarian GPs were observed in the number of GPs contacting patients that were quarantined at home in order to monitor the progression of the disease , and in the number of GPs that preferred to treat patients with mild illnesses that were not linked to suspected cases of COVID-19 by phone or online .Slovenian GPs rated their efforts to control the spread of the virus in the practice  more highly than Hungarian GPs , Hungarian , Italian  and Slovenian  GPs. Furthermore, Austrian GPs reported the decrease in the number of patient contacts  was significantly higher than did Slovenian GPs.German GPs estimated the decrease in the number of patient contacts  to have been significantly higher than did Australian  Australian  and Italian  GPs  to be considerably lower than did Hungarian  GPs . On an ip = 0.006). The biggest difference in their evaluations was in whether the guidelines on how to deal with suspected cases of COVID-19 were sufficiently detailed (CH: 87.3 vs. IT: 32.4%). Australian GPs rated the provision of information to GPs  more highly than Italian (p = 0.001), Austrian  and German  GPs. Between Australian and Italian GPs, the biggest difference in the evaluation was whether they had received guidelines on how to deal with suspected cases of COVID-19 in a timely manner (AU:84.8 vs. IT: 33.8%).Swiss GPs rated the provision of information to GPs  more highly than Italian GPs  than did German , Australian , Slovenian  and Swiss  GPs. On an item level, 92.6% of Italian and 82.7% of Austrian GPs but only 12.9% of Slovenian GPs thought the number of tests that were conducted was insufficient. Furthermore, 10.1% of Austrian and 5.9% of Italian GPs but 81.0% of the Slovenian GPs said that they had adequate access to tests from the beginning of the COVID-19 pandemic.Differences in the evaluation of the effort expended on testing suspected cases varied substantially between countries. Italian , Austrian  and Hungarian . GPs rated the effort expended on testing suspected cases as significantly lower (p = 0.003) Austrian , and Italian GPs . Big differences between Australian and German, Austrian and Italian GPs were found in how they rated the preparedness of their practices to face a pandemic , and whether they had sufficient information on how much protective equipment they would need .Australian GPs rated their preparedness for a pandemic  higher than German  confirmed that a fall in patient contacts was a global phenomenon. Nevertheless, primary care physician consultations remained stable in Australia . A decliGPs in Switzerland and Australia rated the amount of information they had been provided with more highly than other countries, which is probably because they received guidelines on how to deal with suspected COVID-19 cases earlier . As in SThe availability of diagnostic COVID-19 testing varied substantially across countries. Our analyses revealed that GPs from Italy, Hungary and Austria said that in the early stages, access to laboratory testing for COVID-19 in GPs practices was inadequate and rarely carried out. These GPs increasingly demanded that they should be permitted to decide who should be tested and who should not see . Since tGPs in our survey complained that they could not protect their staff, but such complaints were not only heard from GPs, with the British Medical Association, for example, also warning that doctors were at “considerable” risk due to a lack of personal protective equipment , 50. FurOur study has some limitations. Firstly, the questionnaire was developed in a very short time in order that it could be answered when the situation was most acute. Even though we tried to include all relevant topics, some issues may therefore have been missed. Secondly, we could not calculate the response rate because a systematic area-wide survey was not possible in the time frame we permitted ourselves. Nevertheless, the number of responses far exceeded our expectations, especially considering the difficulties that are usually encountered in recruiting GPs for research. Thirdly, as the recruitment process was conducted through regional networks and professional associations, the participants may not have been representative of GPs as a whole. Fourthly, we know that online surveys are not as suitable for the collection of in-depth information as interviews. Fifth, this cross-sectional survey was carried out in the first half of 2020, since when many changes have occurred in pandemic management. For this reason, we also carried out a longitudinal survey in Germany and Austria, which revealed that physicians in primary care have adapted quickly to new situations and have gained experience in telemedicine, enabling them to overcome changes in the delivery of routine health care . Despiten = 4; Switzerland- Germany: n = 4) and countries using different languages , the DIFs are assumed to be due to aspects other than language . The highest number of DIFs could be observed in a comparison between Germany and Slovenia (n = 8)  to investigate the extent of any discrepancies. This analysis investigates whether GPs from different groups  that are being assessed using the same scales  answer individual items in a similar way. The analysis revealed that on two scales  and 16 further items, no DIF could be found, while only one DIF was found in 6 out of 39 investigated items. Since the number of DIFs is comparable in countries where the same language is spoken  .Although general practitioners in different countries were confronted with the same pandemic, its impact differed in a number of aspects . To some extent this may be because countries presumably had different levels of experiences regarding prior pandemics and therefore in how well-prepared they were to face them. This, in turn, may have been reflected in differences in the provision of information and equipment to GPs. Differences in health care systems between the countries may also have had an impact. Knowledge of these differences is important and should be shared in order to be better prepared for future pandemics. It is also essential that primary care representatives are involved in the preparation of structured care plans for future infectious diseases at an early stage.Question: Are there differences regarding management and personal experience of general practitioners in seven countries at the beginning of the COVID-19 pandemics?Finding: Despite facing the same pandemic, there was substantial variation in almost all evaluated dimensions, for which differences in health care systems and experiences with recent pandemics may be responsible.Meaning: GPs should be involved in developing a coordinated strategy to deal with pandemics, which should be communicated to affected health care institutions and populations as early as possible.Study protocol: Detailed information is provided in the Supplement of the Siebenhofer et al. .r et al. . Data seThe 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 Ethics Committee of Bond University, Australia (AS200424). Ethics Committee of Goethe University Frankfurt, Germany (ID 20-619). Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.Conceptualization: ASi, AA, DS-S, KM, SH, HB, MF, ASc, and MG. Data curation: SH and AA. Formal analysis and visualization: AA. Investigation: AA, SH, ASc, AT, EZ, AM, GP, SS, KJ, DS-S, KM, HB, MF, ASi, and MG. Methodology: AA, HB, MF, and ASi. Project administration: DS-S, KM, SH, MF, and ASi. Resources: ASi and KM. Supervision: ASi and MF. Writing—original draft: AA, ASc, ASi, and MG. Writing—review and editing: AT, EZ, AM, GP, SS, KJ, DS-S, KM, AA, SH, HB, MF, ASi, and MG. All authors contributed to the article 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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Anastomotic leakage (AL) is one of the most serious postoperative complications. This study aimed to investigate the predictive value of preoperative body composition for AL in patients with colorectal cancer (CRC).We first reviewed data from 3,681 patients who underwent radical CRC resection 2013–2021 in our hospital, and 60 patients were diagnosed with AL after surgery. We designed a nested case-control study and two controls were randomly selected for each case according to the time and position of surgery. Body composition was measured at the level of the third lumbar vertebra based on computed tomography (CT) images. The risk factors of AL were analyzed by univariate and multivariate analysis. Nomogram was built using binary regression analysis and assessed for clinical usefulness, calibration, and discrimination.In the multivariate analysis, gender, blood glucose, nutrition risk screening (NRS), skeletal muscle area (SMA) and visceral fat area (VFA) were independent risk factors for developing anastomotic leakage after surgery. The prognostic model had an area under the receiver operating characteristic curve of 0.848 . The calibration curve showed good consistency between the predicted and observed outcomes. Decision curve analysis indicated that patients with colorectal cancer can benefit from the prediction model.The nomogram that combined with gender, blood glucose, NRS, SMA, and VFA had good predictive accuracy and reliability to AL. It may be conveniently for clinicians to predict AL preoperatively and be useful for guiding treatment decisions. Colorectal cancer (CRC) is one of the most common cancers of the digestive system. According to statistics, CRC is the third most commonly diagnosed malignancy and the fourth leading cause of cancer-related deaths in the world . CurrentAlthough the surgical techniques for CRC have improved significantly during the past decades, postoperative complications still plague surgeons. Among these complications, anastomotic leakage (AL) remains one of the most potentially life-threatening sequelae in CRC surgery due to its devastating impact on patients' mortality, short- and long-term morbidity, quality of life and survival, with the incidence ranging from 1% to 30% . MoreoveComputed tomography (CT) is a radiographic method commonly used in medical imaging. In terms of its usefulness in body composition measurement, it produces thin cross-sectional high-resolution images that can be processed to differentiate and measure volumes of fat and lean tissue. CT-based multiple body composition parameters have been used in various groups of patients to predict prognosis –12. PrevAlthough almost all patients with CRC underwent abdominal CT examination prior to surgery, assessment of CT examination was limited to assessing tumor stage and the presence of distant metastases. The value of CT images reflecting body composition and patient fitness has not been used in clinical practice. Therefore, our purpose was to develop a helpful nomogram calculating each patient's individual probability based on predictive parameters of epidemiological, surgery-related data and laboratory parameters on the development of AL and examined the predictive value of body composition parameters.A total of 3,681 patients received radical resection of CRC at our center between September 2013 and September 2021. There were 60 patients who were diagnosed with AL. We firstly performed a nested case-control study to identify the risk factors for AL, and two controls were randomly selected for each case according to the time (±1 month) and position (rectum or colon) of surgery. Patients who met the following criteria were included: (1) age ≥18 years; (2) primary colorectal adenocarcinoma confirmed by histopathology; (3) available abdominal CT examination within 2 wk before surgery; (4) no neoadjuvant chemoradiotherapy was performed before surgery. The exclusion criteria were as follows: (1) patients who had definite metastasis before surgery; (2) insufficient clinical information; (3) palliative resections. Finally, 180 patients with pathologically confirmed colorectal adenocarcinoma were included. All surgeries were performed with experienced gastrointestinal surgeons as the principal operator and in strict accordance with the standard surgical procedures. Smoking status was defined as smoking more than 1 cigarette a day for more than a year, regardless of their age at quitting and length of time since they quit. Alcohol status was defined as drinking more than twice a week for more than half of the year.This retrospective non-intervention study was approved by the Ethics Committee of our hospital, and the requirement for informed consent was waived.AL was diagnosed based on clinical and radiologic manifestations: (1) suppurative or intestinal secretions through the drainage tube; or (2) the presence of air or abscess near the anastomotic site detected on CT; or (3) leakage found by X-ray contrast examination.Preoperative CT images of all enrolled patients at the middle level of L3 were extracted from the institutional PACS (Picture Archiving and Communication System). All relevant images were anonymized, transferred to a personal computer, and analyzed using Tomovision's SliceOmatic . The CT HU thresholds were −29 to +150 for skeletal muscle area , −150 toSome data were missing for variables, including white blood cell (WBC), platelet, albumin (Alb), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). We filled in missing data using the technique of multiple imputation by chained equations, which samples imputed values from the posterior predictive distributions of missing data. We assumed data were missing at random. The imputation model was specified on all predictors, outcomes, and dummy variables for study. Five imputations were carried out as this has a relatively high efficiency . We genet-test or Mann-Whitney U test was used to analyze continuous variables according to normal distribution. Shapiro-Wilk test was used to check the normality. Univariate and multivariate analyses logistic regression analysis were used to determine independent risk factors. We carried out an internal verification process with 1,000 bootstrap resamples. The discrimination of the model was calculated using the area under the receiver operating characteristic curve (AUC), and the calibration power was analyzed using the calibration curve. Decision curve analysis (DCA) and clinical impact curves were used to calculate the net benefit. All tests were two-sided, and a value of p < 0.05 was considered to have statistical significance.All statistical analyses were done by SPSS 25.0 software and R-software version 4.1.2. Pearson's chi-square test was used to analyze categorical variables. Student A total of 180 patients  were included in the present study. All patients underwent radical resection of CRC through laparotomy or laparoscopy and 60 patients were diagnosed with AL among them. There were statistically significant differences in gender, WBC, Alb, blood glucose and NRS. The detailed characteristics of these patients are presented in Median and Q1~Q3 of SMA, VFA, SFA, IMFA at the L3 spinal level, resulting from area-based quantification, were provided in p < 0.05 were significantly related to AL, namely gender, WBC, Alb, blood glucose, NRS, SMA and VFA. Before incorporating multivariate regression analysis, we examined these factors for linearity test, multi-collinearity and influential point.According to the results of the univariate analysis in p = 0.0042 was used as the significance level. Our results showed that all the transformed continuous independent variables in the model  have a p-value > 0.0042, indicating that each of them had a linear relationship with the outcome variable , and 2 categorical variables. Therefore, variable .The Variance Inflation Indicator (VIF) and tolerance factor are used to show whether a predictor has a strong linear relationship with the other predictor(s). If the tolerance is <0.1 or the VIF is >10, then there is multi-collinearity. In this case, the variance inflation factor (VIF) for all the independent variables was between 1.024 and 1.109, while tolerance was between 0.902 and 0.977, so there is no multi-collinearity .Cook's distance is used in regression analysis to identify influential data points that may negatively affect your regression model. Any point with a Cook's distance over 4/n  is considered to be an outlier. Our results showed that the Cook's distance for each observation is less than the threshold, indicating that there are no influential data points in the dataset .p < 0.05 were further included in the multivariable model. The results showed that gender, blood glucose, NRS, SMA and VFA were independent risk factors for the occurrence of AL (p = 0.005). Further, blood glucose proved to be a significant independent predictor, AL was more likely to occur in patients with higher blood glucose . Additionally, lower SMA and higher VFA were more likely to develop AL . Concerning NRS, the risk of AL was the highest in NRS ≥5 . On this basis, we established a nomogram  analysis, the area under the curve (AUC) of the nomogram was 0.848 , indicating that the nomogram can predict AL effectively . FurtherR = 0.57, P < 0.001), SMA , SAT , but not with IMF .The results of the correlation analysis of the BMI with CT body composition were shown in AL is one of the most serious complications after CRC surgery, resulting in severe morbidity, prolonged hospital stays, intensive use of medical resources, and increased risk of death. Previous studies have shown that obesity, gender, history of diabetes, etc. are risk factors for the occurrence of AL . HoweverIn our study, gender, blood glucose, NRS, SMA and VFA were independent factors for AL occurrence in CRC. Blood glucose is the most significant influencing factor, and the risk of AL was significantly higher in patients with higher preoperative blood glucose levels. Similar to our findings, the study by Kotagal et al. showed that diabetic (DM) patients had significantly higher rates of postoperative adverse events than non-diabetic (NDM) patients. Even among NDM patients, hyperglycemic patients had an increased risk of adverse events compared with normoglycemic patients. A dose-response relationship exists between blood glucose levels and composite adverse events in NDM patients . The undThe results of correlation analysis between BMI and body composition showed that BMI was significantly positively correlated with VFA, SMA and SAT, but not with IMF. In multivariate analysis, VFA and SMA were independent risk factors for postoperative AL, suggesting that VFA and SMA may be more sensitive predictors of AL than BMI , 36.Nomogram is a graphical representation of a complex mathematical formula , and oneCommonly used nutritional status assessment tools, such as BMI and NRS scores, can only observe the overall status, but cannot obtain individual components of the body, such as regional fat distribution and muscle composition. More and more studies have confirmed that nutritional assessment based on body composition can better reflect the patient's metabolic status and physiological reserve, and may be a decisive factor affecting postoperative outcomes . Almost There are some limitations to our study. First, this is a single-center retrospective study with a small sample size, the established nomogram lacks external validation, and its performance in an external cohort remains to be studied. Second, although we selected controls as representative as possible, selection bias may still exist. Third, we only assessed preoperative body composition and lacked information on postoperative body composition changes and prognostic data. Further, some indicators were meaningful in univariate analysis but not in multivariate analysis, which may be due to the small sample size. Therefore, larger prospective multicenter studies are necessary in order to approve these preliminary results.In conclusion, based on the clinical data and imaging information we collected, we developed a nomogram that can predict AL in patients with CRC. The performance of nomogram was verified by various methods, and the results showed that the nomogram had high accuracy and reliability in predicting AL. Preoperative prediction of AL can help surgeons make appropriate therapeutic decisions in clinical practice.The original contributions presented in the study are included in the article/S-LL and YL: conceptualization and supervision. SX, S-LL, T-YW, and Z-TY: data curation, methodology, and software. SX and Y-KY: writing original draft. S-LL: writing—review and editing. All authors contributed to the article and approved the submitted version.The study was supported by the National Natural Science Foundation of China (Grant No. 81802888), the Key Technology Research and Development Program of Shandong (No. 2018GSF118088), and the General Financial Grant from the China Postdoctoral Science Foundation (No. 2016M592143).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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


N = 771, M age = 14.01 years). A total of two studies were undertaken: (1) the scale was translated into Spanish and its internal consistency, factorial structure and convergent validity were established; and (2) factorial analysis was undertaken to confirm the questionnaire. The results yielded high scores for internal consistency, reliability  and convergent validity. The examination of the underlying nomological network revealed links with positive and negative feelings, anxiety and aggression. According to the Exploratory Factorial Analysis (EFA), the aggregate variance of the factors in the scale was 65.814%, indicating that they can explain variations in aggression levels in adolescents. For its part, the Factorial Confirmatory Analysis (FCA) confirmed the match between the translation and the model, leading to a sustainable model composed by the three factors identified and eight items: χ2 (17) = 30.693; p < 0.001; χ2/gL = 1.805; CFI = 0.968; NFI = 0.837; TLI = 0.944; RMSEA = 0.060, IC del 95% (0.048–0.072). The short scale is easy to understand and quick to complete and is thus considered a useful instrument to assess aggression levels in adolescents.Aggressive beliefs and attitudes are increasingly present in adolescents, and it can be argued that they are a prevalent feature of adolescence. Michel, Pace, Edun, Sawhney, and Thomas’s (2014) original thirty-item scale was later shortened to a more parsimonious eight-item scale (ABA-SF). This study addresses the adaptation and validation of the brief Aggressive Beliefs and Attitudes Scale to Spanish adolescents. The sample comprised a group of Spanish adolescents ( Insults, threats, physical and psychological aggression, humiliation, etc., are increasingly common among adolescents, both in real life and on social media. Aggression prevalence rates vary significantly from study to study . For insThis is linked to an increase in high-risk practices among adolescents, for instance their participation in potentially harmful activities , including physical injuries and exposure to violent behaviour, unsafe sexual practices, and the consumption of tobacco, alcohol and illegal substances ,5. AlthoThis has led to an increasing interest in the study of aggressive attitudes that can affect their behaviour directly, and in the design of effective interventions to prevent them in adult life. In addition, it is likely that those that suffer or display these aggressive behaviours can affect others and, in consequence, their behaviour as well. There are different types of aggressiveness in young people: physical ; verbal (insults); facial (angry expressions on the face); indirect (towards objects of the affected person); and sexual .Most of the existing studies on the factors which affect aggressive behaviour in adolescents are lateral in nature, are result-focused (high risk practices in adolescence), or address proximal factors, that is, those that post-date early infancy . Few studies have examined the link between aggression and aggressive beliefs and attitudes (risk factors) . The aboMany studies have addressed aggressive conduct in adolescents and its associated factors. The role of moral withdrawal as one of the main predictors of aggressive behavior along with other issues such as bullying has been widely described. This would help researchers to understand the processes that leads young people to engage in aggressive behaviors ,13. As aImplicit and explicit cognition are important factors to understand how personality affects behaviour ,21,22,23In addition, explicit social cognitions, in contrast with the implicit, occur at a conscious level, so they are easily accessible to introspection by means of direct methods, such as self-administered questionnaires. However, both, the implicit and the explicit, are needed to explain how personality affects behaviour, and can be used to predict aggression ,35.Despite theoretical advances, no scale existed to measure the relationship between explicit social cognitions and James’s  six aggrThe aggressive beliefs and attitudes questionnaire has demonstrated its psychometric validity, in both its original  and shorThe aims of this study are as follows: (1) to evaluate the possibility of translating the aggressive beliefs and attitudes questionnaire in its short format  into SpaThe only hypothesis of the study is that the aggressive beliefs and attitudes questionnaire is a valid instrument to measure this construct in Spanish adolescents.In order to select the sample, schools were contacted telephonically, and a list of participating schools was drafted. The principal investigator explained the main objective of the study to the participants when the questionnaires were delivered, emphasizing the importance of responding to all issues. Participants had 45 min to complete the questionnaires and the informed consent form. Participants were informed that the information would be treated anonymously and confidentially. The data were collected in October and November 2021.The original English version was translated into Spanish by two bilingual translators (native Spanish speakers), with experience in the translation of scientific literature in the field of psychology. The translations were compared and examined by the research team, and a first Spanish draft was agreed.A psychologist with professional experience in English-speaking countries assessed the conceptual equivalence, clarity and naturalness of items and responses in this first version, and a second version was drafted with the necessary corrections.The new version was circulated to psychologists working with adolescents for comment.The second Spanish version was translated back into English by a bilingual translator (native English speaker).A pilot test with 50 secondary school students was undertaken in order to test the clarity of the questionnaire, the response time and the suitability of the responses.Based on these preliminary results, a third version of the scale was drafted and tested to assess its suitability to Spanish adolescents.The translation of the Aggressive Beliefs and Attitudes Scale into Spanish followed the standard process of reverse translation set out by Muñiz, Elosua & Hambleton , which cs.d. = 1.67). Parents and participants were asked to sign an informed consent form, and all the ethical considerations set out in the Declaration of Helsinki were met; the study was endorsed by the Ethics Committee of research group OPIICS (S46_20R), University of Zaragoza; all ethical criteria for research with human beings were met . The sample was found to be representative of the province of Zaragoza, with a 99% confidence level. The study was designed as an ex–post facto retrospective study [The sample comprised 771 secondary school students from Zaragoza (Spain)—361 men (49.42%) and 451 women (50.58%)—all of whom were volunteers . The aveve study . ResultsTo evaluate the variables studied, the following instruments were administered: Aggressive Beliefs and Attitudes Questionnaire-Short Form , SPANE positive and negative feelings scale  and Zuckerman–Kuhlman Personality Questionnaire ZKPQ-50-CC . Respondents were asked to fill out a biographical information form and the survey to assess the variables studied. At this point, single method bias is regularly mentioned as a limitation of the survey design. However, this was mitigated by the limited number of items contained in the research questionnaire, and furthermore, the questions were worded in a clear and concise manner . FurtherAggressive Beliefs and Attitudes Questionnaire-Short format [t format .Explicit aggressive beliefs and attitudes were originally assessed through a 30-item scale developed and validated by Michel et al. , and lat2.SPANE positive and negative feelings scale [gs scale .The positive and negative feeling scales evaluate the feelings experienced by the participant in the four weeks prior to the survey. They comprise 12 items, six for positive affects (PA) and six for negative feelings (NA), in two Likert scales. Respondents must refer to their feelings at the time of filling in the questionnaire, and responses range from 0 (absence of emotion) to 5 (frequent presence of emotion). All the SPANE items score from 1 to 5, where 1 means ‘very rarely or never’ and 5 means ‘often or always’. Positive and negative scales are examined separately, owing to the partial independence of separability of both types of feeling. The aggregates can range from 6 to 30 , and from −6 to −30 (negative SPANE-N). The aggregate of both scales (SPANE-B) can range from 24 to −24, which reflects the affective balance. The scale yielded a reliability score of α = 0.89; Ω = 0.90 for positive feelings and α = 0.86; Ω = 0.88 for negative feelings. 3.ZKPQ-50-CC [PQ-50-CC .It is a self-administered questionnaire used to assess personality. It comprises 50 items with true/false answers. It assesses the different dimensions of personality, using alternative five as factors: (1) Impulsive sensation seeking: willingness to run risks for excitement and new experiences; (2) Neuroticism-anxiety: emotional instability, tension, concern, phobias and/or fears; (3) Aggression-Hostility: verbal aggression, rudeness, antisocial behaviour and rancour; (4) Activity: need for action, inability to stay idle; and (5) Sociability: number of friends and time spent with them, preference for company over solitude. The scale of Aggression-Hostility (Agg-Host) is a combination of willingness to express oneself aggressively and rudeness, disregard, antisocial behavior, revenge and malice, or from the other pole, willingness to be pleasant and cordial and have a friendly treatment with people. In terms of internal consistency, the ZKPQ-50-CC yields an α score between 0.78 and 0.82, and its convergence validity is regarded as adequate. In this study, α values for the different factors ranged from 0.77 to 0.83; and Ω values from 0.79 to 0.84.Study 1. Descriptive statistics were undertaken to establish the sociodemographic characteristics of the data: averages and standard deviations for the quantitative variables and percentages for the nominal variables. Reliability and validity were calculated in order to establish the questionnaire’s psychometric properties. In terms of reliability, Cronbach’s alpha [’s alpha  was calcStudy 2. The following goodness of fit indicators were examined: chi-square (χ2); comparative fit index (CFI); Tucker–Lewis’s index (TLI); and Root Mean Square Error of Approximation (RMSEA) with a 90% confidence interval. Based on typical criteria, CFI and TLI values of 0.90 and 0.95 are regarded as an indication of good and excellent fit, respectively, while RMSEA values below 0.08 and 0.06 are considered indicative of acceptable and excellent fit, respectively.The results are divided into two groups, one for each phase of the study. The following results correspond to Study 1: construct validity; internal consistency analysis; convergent validity; and Exploratory Factorial Analysis (EFA); the results for Study 2 correspond to the Confirmatory Factorial Analysis (CFA).The aim of the study was to validate the Aggressive Beliefs and Attitudes Questionnaire, in its short format , with adp < 0.001). The Kaiser Meyer Adequacy (KMO) test yielded a value of 0.716, suggesting that the correlations between pairs of variables can almost be totally explained by the remaining variables. All Measures of Sampling Adequacy (MSA) values were above 0.75. These values recommended the correlation matrix to be subject to factorial analysis. As shown in The principal components analysis with Oblimin rotation method was chosen after the viability of the factorial analysis was established, with the following criteria: the correlation matrix yielded a large number of correlations (85.1%) with values in excess of 0.30 (determinant = 0.002), and Bartlett’s test of sphericity indicated that variables were not independent  = 30.693; p < 0.001; χ2/gL = 1.805; CFI = 0.968; NFI = 0.837; TLI = 0.944; RMSEA = 0.060, IC del 95% (0.048–0.072).Concerning the goodness of fit, the various fitness indexes returned adequate values, suggesting that the proposed model for the factorial structure of the scale is sustainable: χ2 value was 30.693 with 17 degrees of freedom; and RMSEA yielded an optimum fit, with an index of 0.060 (Lo = 0.048–Hi = 0.072). These results .The confirmatory factorial analysis model had three factors. This is an a priori structure, which means that the results of the model are entirely confirmatory. The indices returned a reasonably adequate fit; the χ results  suggest  results  was signThe aim of this study was to explore the use of the ABA-SF scale to measure aggression in adolescents.The study validated the ABA-SF scale, following a protocol that included the translation, adaptation, and psychometric evaluation of the questionnaire. The article presents the protocol followed to ensure the cultural and linguistic equivalence of the original scale and the adapted version, and the tests undertaken to establish its internal validity and factorial structure.The results of factorial analysis yield a three-dimension structure and high saturation in all items, which is a reflection of good internal consistency (0.851), similar to those returned by the questionnaire in earlier studies ,36. The results clearly showed that the factors of the ABA-SF scale present an accumulated variance of 65.81%. Other variables also support the validity of the questionnaire, such as the high consistency attested between elements of the scale, which is indicative of great reliability.In addition to exploring the underlying factorial structure, reliability indices for each subscale, similar to those returned by the original model, were also established. It can thus be confirmed that the questionnaire keeps consistent when used with adolescents. The three-factor structure was confirmed with empirical data. The three underlying factors are significantly correlated: Victimization-Derogation (r = 0.771 **); Victimization-Retribution (r = 0.678 **); Retribution-Derogation (r = 0.403 **); all of them are above 0.4, and two of them can be regarded as very strong correlations.In a similar way, the analysis of the correlations of the scale with those that assess personality and emotions reveals that victimization, which reflects the individual’s tendency to feel victimized by the powerful, only correlates with neuroticism-anxiety, which suggests that adolescents that feel victimized are less prone to anxiety, probably reflecting an inclination to surrender to adversity. The derogation factor, the aggressor presenting the victim as deserving of the aggression, correlates negatively with anxiety, aggression and positive feelings, and positively with negative feelings. Finally, retribution, which measures the tendency to give priority to reprisals over reconciliation, correlates negatively with anxiety, aggression and positive affects—anxiety, aggression and positive affects levels drop after the reprisal—and positively with negative feelings. Therefore, the study has found preliminary evidence for correlations between the ABA-SF, BFQ-NA  and SPANE (feelings) questionnaires. Moreover, our results satisfactorily responded to the underlying theoretical model and returned high internal consistency and validity values. As noted, social cognitions, both implicit and explicit, are important to understand the influence of personality on behaviour, and the ties of these constructs with social skills, self-esteem, emotions, sentiments, etc. ,48,49,50These results must be interpreted in light of the limitations of the study. Although the sample is statistically significant, it should be expanded to include other population segments, in which different correlations between aggressive beliefs and attitudes and other constructs may be attested. It may also be accurate to study the difference between Spanish-speaking countries in terms of their conception of violence within their culture. It is also advisable to undertake longitudinal studies that cover the evolution of these constructs over a longer time period. In addition, aggressive behaviour is often compounded with family, school and social contextual factors, and this relationship should be explored further. The questionnaire should also be used to analyze population groups with special characteristics, such as adolescents suffering from anxiety or autism, which present a different profile in terms of social cognitions, both implicit and explicit ,57,58. The main conclusion of the study is that the ABA-SF is a valuable tool to measure aggressive beliefs and attitudes in adolescents; it is psychometrically efficacious, its factorial structure is solid, and it is internally consistent; in addition, it is quick and easy to implement. Although further research is necessary, these preliminary results indicate that the ABA-SF questionnaire is a useful tool to measure aggression in adolescents.The results of the research endorses the validity of the ABA-SF as a short, easy to implement and understand, and adequate for the measurement of the target constructs, research tool. Besides, the factorial structure model proposed is sustainable; three factors and eight items were identified. The translated ABA-SF yielded satisfactory coefficients for each of the three factors identified in its factorial structure. In our perspective, this further demonstrates the validity of the scale in adolescents, and confirms that the Spanish version faithfully replicates the theoretical structure of the original.


Regardless of gender and age groups, significant differences were observed between overweight/obese and normal-weight adolescents in SBP (p < 0.001), DBP (p < 0.001 to 0.009), triceps skinfold (p < 0.001), and abdominal perimeter (p < 0.001). Conclusions: Our findings revealed higher SBP, DBP, triceps skinfold, and abdominal circumference in overweight/obese adolescents compared to normal-weight adolescents in both genders.Background/Objective: The greatest anthropometric and physiological changes occur during adolescence. Assessment of growth patterns is necessary to prevent future health risks. Aims: To describe the values of glycosylated haemoglobin (HbA1c), systolic (SBP) and diastolic (DBP) blood pressure, triceps skinfold, and abdominal circumference according to gender and age (between 12 and 17 years), as well as explore the differences between body weight conditions. Methods: A descriptive cross-sectional study was carried out, including 4130 adolescents between 12 and 17 years old. SBP and DBP, HbA1c, triceps skinfold, and abdominal circumference were evaluated. Results: Significant differences were observed between males and females for HbA1c ( Cardiovascular disease (CVD) is the leading cause of death worldwide. CVD risk factors include high cholesterol, blood glucose levels, and obesity . As blooIn recent years, the prevalence of hypertension has increased markedly in adolescents. At this stage, elevated blood pressure levels are often influenced by weight or genetic factors . Obesity2.Anthropometric has been widely used to screen for cardiometabolic risk in children and adolescents, mainly due to its low cost, ease of administration, and non-invasive nature . FurtherAlthough BMI could be the measure used to define levels of overweight and obesity, it should be considered an indicator of weight but not adiposity  as it doWaist circumference is a measure of central or visceral adiposity that may be a good indicator of adiposity, as well as a better predictor of elevated BP and risk of suffering cardiovascular diseases (CVD) in children than BMI ,20,21,22The greatest anthropometric, hormonal, and psychological changes occur between 12 and 18 years old, coinciding with adolescence. At this stage, being overweight was linked with a superior prevalence of high-level diastolic blood pressure (DBP), cholesterol, triglycerides, and glycosylated haemoglobin (HbA1c) in fasting blood ,27. For Therefore, knowledge of anthropometric development in pre-adolescent and adolescent stages is essential to prevent the appearance of health risks. Thus, this study aims to examine the values of HbA1c, DBP, SBP, triceps skinfold, and abdominal circumference according to gender and age in adolescents between 12 and 17 years old, as well as explore the possible differences between different body weight conditions .A descriptive cross-sectional study was conducted during a 14-month period from October 2007 to December 2008. A multistage stratified sample was utilised. The units of the successive stages were districts, health areas, educational centres (IESO), city typology , academic year, gender, and students (surveyable). The sample was selected from forty secondary schools.Forty secondary schools from the Extremadura region (Spain) take part in this study. Schools were randomly selected to ensure the selection of a representative sample from all health areas in Extremadura. Thus, 4130 teenagers (2128 boys and 2002 girls) between 12 and 17 years old [14.25 (1.48)] were evaluated. Individuals had to meet the following eligibility criteria: (1) age between 12 and 17 years old; (2) live in the Autonomous Community of Extremadura; (3) be authorised by their parents or legal guardians; and (4) to be agreeing to join in the study.The procedures of the present study were approved by the Bioethics and Biosafety Committee at the University of Extremadura, in agreement with the Declaration of Helsinki guidelines (reference code 11/2006). Both participants and their respective parents or legal guardians signed an informed consent form accepting their participation in the study.www.ine.es, consulted on 16 June 2021) ); normal (=1 SD); overweight (>1 SD to ≤2 SD); and obese (>2 SD) .Blood pressure was assessed in a sitting position with a manual sphygmomanometer and a stethoscope. It was measured at three different times: at the beginning, in the middle and at the end of the interview. The mean of the second and third records was considered to define high blood pressure (HBP). Individuals with SBP > 140 and/or DBP > 90 mmHg were classified as hypertensive according to the 2018 European Society of Cardiology (ESC)/European Society of Hypertension (ESH) guidelines . In addiSBP and DBP were assessed with an oscillometric device  following the Fourth Report on the Diagnosis, Evaluation and Treatment of High Blood Pressure in Children and Adolescents recommendations . The cufThe procedure was explained to each participant, and they were asked not to speak during the measurement. The patients remained seated, with their backs supported and feet placed on a flat surface, in a comfortable environment in a resting position for five minutes. The arm was uncovered up to the shoulder and rested on a horizontal surface at the heart level. Beforehand, the circumference of the arm was measured to meet the requirement of using the most appropriate cuff, with a width of 40% and a length of 80–100%. The arm cuff was centred on the brachial artery without over-tightening or leaving it too loose. The lower edge of the cuff was placed 3 cm above the antecubital fossa.Subsequently, the maximum inflation level was determined using the Osler manoeuvre to avoid pain or discomfort during the measurement. The stethoscope was located over the brachial artery with the entire surface in contact with the skin, and then the arm cuff was inflated rapidly and steadily to the predetermined level, releasing air from the chamber at a rate of 2 mmHg per beat. SBP was determined when two continuous beats appeared (Korotkoff phase I) and DBP at the time when the sounds disappeared (Korotkoff phase V). An additional 10 mmHg was examined after the last beat to confirm the disappearance of the beats.The waist circumference  was deteHbA1c was quantified by high-performance liquid chromatography  . Descriptive statistics were executed for all parameters. Data are presented as means and standard deviation (SD).p ≤ 0.05. Additionally, Hedge’s g effect size with a 95% confidence interval was also estimated to compute the magnitude of comparisons. Effect size thresholds were the following [Kolmogorov–Smirnov and Levene’s tests were applied for analysing the normality and homogeneity of data, respectively. After that, the Mann--Whitney U test was employed to analyse between-gender and between-bodyweight condition comparisons in all dependent variables due to the non-parametric distribution of data previously confirmed. The alpha level was fixed at ollowing : >0.2, sp < 0.001), SBP (p < 0.001), triceps skinfold (p < 0.001), and abdominal perimeter (p < 0.001), independently of the age group. There were also meaningful differences in DBP between gender in the 15–17 age group. However, no differences were detected for DBP in the 12 to 14 years old group (p = 0.272).p < 0.001), DBP (p: <0.001 to 0.009), triceps skinfold (p < 0.001), and abdominal circumference (p < 0.001). In contrast, no differences were observed between bodyweight conditions for HbA1c in males (p = 0.397 to 0.459) nor females (p = 0.429 to 0.563), independently of the age group.The main findings show significant differences in SBP, DBP, triceps skinfold, and abdominal circumference between overweight/obese and normal-weight adolescents. However, no differences are observed for glycosylated haemoglobin levels and DBP in children aged 15–17 years. Thus, overweight/obese children have higher SBP and DBP levels (except for those aged 15–17 years), a higher triceps skinfold, and a larger abdominal circumference. Similarly, previous investigations have also shown an association between anthropometrics and blood pressure in childhood ,31. HypeThese findings are clinically relevant, as obesity and hypertension are associated with high-risk cardiovascular disease, increasing related morbidity and mortality in adulthood ,35. HighEstimating the prevalence of abdominal or central obesity is becoming increasingly common. In fact, several studies have shown that children with excess abdominal fat are also at increased risk of having a more atherogenic lipid profile, higher blood pressure, greater carotid intima-media thickness, and even metabolic syndrome ,40. TherTriceps skinfold thickness was also significantly related to obesity/overweight and normal weight in children, independent of age and gender. Skinfold thickness measurements represent an indirect measure of subcutaneous adipose tissue and are used to estimate total body density to derive body fat percentage . IncreasPrevious findings  suggest On the other hand, due to boys having higher values for SBP, abdominal circumference, and glycosylated haemoglobin (in the 12–14 years group), higher values for DBP (15–16 years group), and girls having higher tricipital skinfold measurement in these ages, these results suggest the presence of gender differences in development from preadolescence to adolescence. The difference in parameter values could be due to the difference in growth between boys and girls.The peak prevalence of overweight in girls tends to occur almost 2 years earlier than in boys (10 vs. 12 years), coinciding with the age of onset of pubertal development, one of the critical moments of body fat gain . Gender Abnormal fat gains during puberty may also reflect an increased risk of developing cardiovascular disease .One of the major limitations of this study was the lack of consideration of diet and eating habits or physical activity levels, which are key factors in glycaemic control.The main strength of the study lies in the representativeness of the sample by gender and age (4130 adolescents aged 12–17), which allows for the provision of relevant data. However, this study has several limitations. The study lacks data for children under the age of 12, so it would be interesting to study this trend at the youngest age. There is a lack of equity in the percentages of obese/overweight and normal-weight boys and girls. In addition, follow-up data are not available, so its cross-sectional design does not allow for establishing causality, and given that this study population covered a well-characterised cohort of children from Extremadura, that may limit the generalisability of the outcomes to other populations.Therefore, future studies focusing on the monitoring of HbA1c, blood pressure, triceps skinfold, and abdominal circumference could be studied to determine the existence of causality between the data and age and gender. In addition, it would be interesting to include children’s ages in future studies to know the evolution of these parameters from childhood to adolescence.Considering our findings, we conclude that adolescent boys have greater SBP, HbA1c, and waist circumference than girls. However, adolescent girls present a higher triceps skinfold. Furthermore, overweight/obese adolescents show higher SBP, DBP, tricipital skinfold, and abdominal circumference compared to their normal-weight counterparts in both gender, except for DBP in boys aged 15–17 years. Moreover, the positive relationship between blood pressure, tricipital skinfold, and abdominal circumference in overweight/obese children may highlight the need to use anthropometric measures to assess body fat distribution and cardiovascular risk to develop effective strategies to reduce cardiovascular risk in this population.