Metschnikowia, Hanseniaspora, Torulaspora, Debaryomyces, Meyerozyma, and Rhodotorula. These yeasts were present in all vineyards studied but with varying loads depending on the valley sampled. No identical fingerprints were observed; however, similarities and differences could be observed among the microbial profiles of each valley. A co-variation in the loads of Metschnikowia and Hanseniaspora with latitude was observed, showing high loads in the Casablanca and Itata valleys, which was coincident with the higher relative humidity or rainfall of those areas. Non-conventional yeasts were also isolated and identified after sequencing molecular markers. Potentially good aromatic properties were also screened among the isolates, resulting in the selection of mostly Metschnikowia and Hanseniaspora isolates. Finally, our results suggest that microbial terroir might be affected by climatic conditions such as relative humidity and rainfall, especially impacting the load of non-conventional yeast. In this study, the microbial fingerprint for yeast in Chilean vineyards is reported for the first time revealing an opportunity to study the contribution of this assembly of microorganisms to the final product.In this study, the presence of non-conventional yeast associated with vineyards located between latitudes 30°S and 36°S was examined, including the valleys of Limarí, Casablanca, Maipo, Colchagua, Maule, and Itata. The microbial fingerprinting in each valley was examined based on the specific quantification of yeast of enological interest. Grape–berries were sampled to evaluate the presence and load of non-conventional yeast with enological potential, such as  Saccharomyces and non-Saccharomyces. The latter category includes a wide array of different genera, also termed non-conventional yeasts. Saccharomyces has a high fermentative capacity and predominates during alcoholic fermentation  with sugar content around 23° Brix. The vineyards were located between latitudes 30°S and 36°S, representing the Limarí, Casablanca, Maipo, Colchagua, Maule, and Itata valleys. Approximately 1 kg of grapes was obtained from at least five plants of each vineyard, as detailed in ® bags and transported to the laboratory in a coolbox containing ice pads. Once in the laboratory, each sample was transferred into a fresh Stomacher® bag, in which the berries were pressed by hand for 3 min. Then, 5 mL of juice was separated for culture analysis, while 5 mL was centrifuged at 12.000 × g for 15 min, and the pellets obtained were frozen at -20°C until DNA extraction.The samples were loaded into sterile Stomacher-2 to 10-4) of each sample (0.1 mL) were plated on YEPD agar medium  with 25 ppm of cycloheximide (Merck) and incubated at 28°C for 2 days. Where possible, 4–6 representatives of each colony-morphology were isolated from plates with ≤200 colonies and purified through two rounds of streak plating onto fresh agar plates. In addition, unique but infrequent colonies that were observed on plates with >200 colonies were also isolated. The isolates were maintained in a cryobank at -80°C.Samples for the isolation and identification of yeasts were taken from the juice samples described above. Several decimal dilutions , with vigorous agitation. A 20 μL aliquot of 20 mg/mL lyticase (Sigma) was added to the samples, which were subsequently incubated at 37°C for 20 min. Then, the samples were treated with 2.5 μL volume of 20 mg/mL Proteinase K (Merck) incubated at 37°C for 45 min. The Power Soil DNA Isolation Kit  was used for DNA extraction according to the manufacturer’s instructions.g for 5 min. The pellets formed were washed with TE-NaCl  and centrifuged at 5.000 × g for 5 min. Subsequently, a 20 μL volume of 20 mg/mL lyticase (Sigma) was added to the samples, which were subsequently incubated at 37°C for 20 min. Finally, the samples were treated with 2.5 μL volume of 20 mg/mL Proteinase K (Merck) incubated at 37°C for 45 min. The Power Soil DNA Isolation Kit  was used for DNA extraction according to the manufacturer’s instructions. All the DNA obtained was froze at -20°C until processed.In the case of yeast isolates, each of the colonies selected was suspended in 200 μL PBS, with vigorous agitation, followed by centrifugation at 5.000 × Saccharomyces) were done by ITS 5.8S rRNA and D1/D2 sequence. The ITS 5.8S-rRNA were amplified using primers ITF1 (5′-TCCGTAGGTGAACCTGCGG-3′) and ITF4 . Isolates were identified and respective sequences were deposited in GenBank (KU350312–KU350496).The identification of yeast isolates  using primers and programs described in 2O. All of the amplifications were carried out in optical-grade, 96-well plates, AriaMx real-time PCR System (Agilent Technologies). All samples were analyzed in triplicate. Yeast load were compared using grouped analysis performed with GraphPad Prism version 6.00 for Mac .PCR amplification was performed in 10 μL of mix containing 1 μL of DNA 0.5 pmol/μL of each respective primer, 8 μL of LightCycler 480 SYBR Green I Master (Roche) and 1 μL of Milli-Q sterile H4. Phylogenetic and molecular evolutionary analyses were performed using MEGA software, version 6.0 Beta. The phylogenetic tree for 5.8S ITS and D1/D2 domain part 26S rRNA gen were constructed by UPGMA (unweighted pair-group method with arithmetic mean) method. The evolutionary distances were computed using the Maximum Composite Likelihood method, using the Mega 6 (version 6.0) software package obtained from the website5.The nucleotide sequences of the 5.8S ITS rDNA region and D1/D2 domain part gen 26S rRNA were compared with those available in the GenBank database using the BLAST method in order to investigate their approximate phylogenetic affiliation, and their sequence similarities were determined at the National Center for Biotechnology Information, USA 4. PhylogTable 3) supplemented with 20 mg/L of SO2 was inoculated to a final concentration of 108 cells/mL. Micro-fermentations were conducted at 18°C with stirring in an orbital shaker at a rate of 150 rpm. The evolution of the alcoholic fermentation was evaluated by monitoring weight loss every two days. At the end of the fermentation, the concentrations of glucose/fructose were measured using enzymatic kits (Boehringer Mannheim), according to the manufacturer’s instructions.With the yeast isolates obtained from each of the grape–berry samples, a series of micro-fermentations were conducted for an initial assessment of the fermentation capacity and aromatic attributes of the isolates. Fifty milliliters of synthetic must , in individual booths. The sensory analyses were carried out by olfactory evaluation in the Sensory evaluation laboratory and all panelists were winemakers belonging to the Enology Department, Universidad de Chile. Terpenes, thiols, and higher alcohols were represented as aromatic descriptors such as “fruit” and/or “flower” and were order as aromatic groups: fermentative, tropical fruit, citrus fruit, stone fruits, berries, flower, dried fruit, cooked fruits, and sweet aromas. The sensory panel first smelled several fresh aromatic references to choose those samples which best fitted their aroma. The strategy for data analysis was a descriptive method (attribute score versus frequencies of citation). The number of times each attribute was cited as negative frequency and positive frequency for each sample were counted up. Once all data were collected, the list of yeast was ranked according to their citation frequency to identify the most relevant attribute of each fermented product.6 and Red Agroclimática Nacional7 databases. Data were collected from seven different weather stations representing each vineyard. Monthly measurements were extracted for average temperature, maximum temperature, minimum temperature, rainfall, and average relative humidity during 2015 .Monthly weather data were extracted from the Agromet INIATable 1), screening for Saccharomyces and six non-conventional yeast genera Torulaspora, Hanseniaspora, Metschnikowia, Rhodotorula, Debaryomyces, and Meyerozyma. All of these yeast were present at different loads depending on the examined samples. The yeast population  was represented for each valley and it revealed the microbial fingerprint for each area, ordered from north to south . No identical fingerprints were observed, however, similarities and differences could be observed among the microbial profiles of each valley. For example, Hanseniaspora and Metschnikowia were present in Limarí, Casablanca, and Itata valleys at comparatively high loads with respect to Maipo and Maule valleys. Interestingly, the following three sets of yeast showed load patterns that were similar across valleys: Hanseniaspora and Metschnikowia; Torulaspora, Saccharomyces, and Meyerozyma; and Rhodotorula and Debaryomyces . The vineyard samples with the highest yeast load were the ones collected in Casablanca Valley, with a dominant presence of Hanseniaspora and Metschnikowia, with 107 yeasts per gram of fruit. Torulaspora/Saccharomyces/Meyerozyma showed a similar population trend along latitude, with high loads in Casablanca (105 yeasts per gram), lower toward the South (about <103 yeasts per gram), with a slight increase for the Itata vineyards. In contrast, the population load of Rhodotorula and Debaryomyces presented a similar behavior along latitude, maintaining loads between 103 and 105 yeasts per gram of fruit.A total of twenty-five samples of grape–berries from six Chilean viticultural areas were analyzed . The high population loads of Hanseniaspora/Metschnikowia observed in the Casablanca and Itata valleys were coincident with the highest relative humidity and rainfall observed for those valleys, respectively. Similarly, the load pattern of Torulaspora/Saccharomyces/Meyerozyma could also be linked with the types of climate variations. They showed the highest load in valleys with low rainfall , but their load was reduced in valleys with high rainfall . Finally, Rhodotorula/Debaryomyces seemed to be independent of relative humidity and rainfall, and their load might be linked to the temperatures observed in the valleys.To try to explain the differences observed in population patterns depending on vineyard location, the relative humidity, rainfall, and temperatures of these areas were explored  and Saccharomyces (2.7%). These results showed an important presence of non-conventional yeast isolates in Chilean valleys. The dimorphic Aureobasidium (24%) black yeast-like fungus was widely distributed in the Maipo and Maule valleys. The predominant non-conventional genera were Metschnikowia (21%), Hanseniaspora (18%), and Rhodotorula (13%). Minority genera included Cryptococcus (6%), Hyphopichia (2%), and Candida (2%). Other isolates corresponded to Lachancea, Zygosaccharomyces, Sporidiobolus, Pichia, Meyerozyma, Debaryomyces, and Torulaspora, which represented less than 1% as a group.More than 200 yeasts were isolated, and the results are summarized in Supplementary Figure S1 shows the tree generated by the D1/D2 domain of the partial 26S rRNA sequences from Metschnikowia isolates, in which 16 different phylotypes can be observed, indicating the high genetic diversity of Metschnikowia. On the other hand, the analysis of Rhodotorula and Hanseniaspora based on D1/D2 sequences  showed limited genetic diversity, with four phylotypes for Rhodotorula and six for Hanseniaspora. The observed diversity of phylotypes seems to be randomly distributed among vineyards rather than corresponding to the predominance of specific genotypes depending on the geographical areas.Among the total samples isolated, 61 yeast sequences were included in a phylogenetic analysis. 2. Micro-fermentations were performed in synthetic must, which was supplemented with metabisulfite to address this criterion. Only 106 isolates were able to tolerate metabisulfite. Then, the enological potential of these isolates was evaluated based on their ability to produce desirable aromas during micro-fermentation.To select non-conventional yeast with enological potential, all of the strains were tested for their resistance to SOTable 4). These isolates corresponded to Metschnikowia , Hanseniaspora , Rhodotorula , Hyphopichia , Candida , Lachancea , Pichia , Debaryomyces , and Citeromyces . The main aromatic attribute in Debaryomyces was dried fruits; in Hanseniaspora isolates were stone, tropical fruits, and sweet aromas; in Pichia isolates were tropical fruits and berries. Interestingly, several Metschnikowia isolates offered different attributes such as fermentative and sweet aromas, tropical, and stone fruits.The attributes most frequently observed in the micro-fermentations were those belonging to the fruity, flowery, and fermentative aromas. Nineteen non-conventional yeasts were selected based on descriptive aromatic profiles . Other isolates can be considered as poor fermenters, with less than 50% sugar consumption.The fermentation abilities of the isolates were also tested. This study evaluated the load and diversity of non-conventional yeasts in Chilean vineyards and was the starting point of a study whose final goal is take advantage of the native microbiome by selection of local strains with interesting enological properties. Similarly to the animal microbiome, the plant microbiome may have important roles for their host, such as improving the availability of organic matter and preventing the growth pathogens through competition for space and nutrients . StudyinThis study has covered a wide viticultural region (from 30°S to 36°S latitude approximately) that included the most important Chilean valleys and described the microbial fingerprinting based on the yeast of enological interest by using a culture independent approach. In contrast, Metschnikowia/Hanseniaspora, Torulaspora/Saccharomyces/Meyerozyma, and Rhodotorula/Debaryomyces that were observed along latitude and associated with relative humidity, rainfall and temperature, respectively. These observations are consistent with studies by Metschnikowia and Hanseniaspora. Similarly, Itata valley showed the highest rainfall during the harvest period, which coincided with a high load of Metschnikowia/Hanseniaspora. Recent studies by Metschnikowia/Hanseniaspora in Casablanca valley, located near to the coast, may be related to relative humidity, as this parameter is highest due to the pronounced maritime influence. Those reports showed that rainfall and relative humidity favored a prevalence of Hanseniaspora and Metschnikowia yeast on the grape–berries, which could explain the co-variation observed in the Chilean valleys. Contrasting results were obtained for Torulaspora/Saccharomyces/Meyerozyma, which seems to vary inversely with rainfall. However, these results have not been reported previously and demand more research efforts to define the negative influence of the rainfall on the load of those yeasts.Additionally, our results allowed us to identify co-variations in the loads of Hanseniaspora, Metschnikowia, and Aureobasidium were the main genera present on grape–berries in all of the vineyards studied in Chile. These observations are consistent with the findings reviewed by Candida, were the main ones present on grape–berries examined in Spain, Canada, and Argentina. Molecular biology techniques provide a simple and rapid method to differentiate yeasts based on their genetic background (Metschnikowia isolates from different Chilean valleys showed that the high genetic diversity was related to 26 polymorphic sites that generate 16 phylotypes. These results agreed with the findings of Metschnikowia species.Based on the culture dependent approach, our results showed that ckground . The modckground . The comSaccharomyces in different wines. Hanseniaspora isolates in Chardonnay produced a uniquely fruity character, such as banana, pear apple, citric fruits, grape fruit, and guava. Metschnikowia has been used in base wine for sparkling wine production, improving the aromatic profile by increasing smoky and flowery notes (Debaryomyces increased the concentrations of citronellol, nerol, and geraniol, which resulted in floral and citrus-type aromas in wine (Debaryomyces isolate obtained in this study showed a different property, improving dried fruit aromas. These studies have demonstrated that non-conventional yeast can be selected based on their ability to produce aromatic secondary metabolites that contribute to improving the quality of wine, which is presently a very important area of applied interest in oenology.Non-conventional yeasts influenced the wine aroma: terpenes, thiols, esters, and higher alcohols are the most typical aromatic compounds that contribute to the enhancement of sweet-fruity aromas in wines . Most ofry notes . In anot in wine ; howeverHanseniaspora, Metschnikowia, and Debaryomyces genera. Therefore, our isolates might influence the wine quality through the release of several aromatic compounds, such as esters, higher alcohols, acids, and monoterpenes. Thus, further research to analyze the potential of these strains is underway to select new starter presentations for the wine industry. Furthermore, these non-conventional yeasts reveal the potential of the yeast microbiome to contribute to the complexity and typicality of the wine and conferring the aromatic profiles of specific regions. This possibility is another aspect that is currently being developed in our laboratories.In our study, we focused on the selection of different non-conventional yeasts as a basis for the selection of interesting strains to be used by the wine industry. In our hands, the most promising yeast strains corresponded to the Conceived and designed the experiments: CJ, VL, AM, and JR. Performed the experiments: CJ and JR. Generated and analyzed the data: CJ, VL, AM, and JR. Wrote the paper: CJ, VL, AM, and JR.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.


The present study contributes to the discussion on the automaticity of semanticprocessing. Whereas most previous research investigated semantic processing atword level, the present study addressed semantic processing during sentencereading. A dual task paradigm was combined with the recording of event-relatedbrain potentials. Previous research at word level processing reported differentpatterns of interference with the N400 by additional tasks: attenuation ofamplitude or delay of latency. In the present study, we presented Spanishsentences that were semantically correct or contained a semantic violation in acritical word. At different intervals preceding the critical word a tone waspresented that required a high-priority choice response. At short intervals/hightemporal overlap between the tasks mean amplitude of the N400 was reducedrelative to long intervals/low temporal overlap, but there were no shifts ofpeak latency. We propose that processing at sentence level exerts a protectiveeffect against the additional task. This is in accord with the attentionalsensitization model , which s This is reflectedin an increase of naming latencies for incongruent  compared to congruent trials. The usual interpretation for thisfinding suggests that during access to a word form, activation automatically spreadsto the semantic information encoded by that word. This mechanism does not requireattention or awareness  is very useful in investigating online-processing of linguistic material. TheN400 component of the ERP is generally believed to be a modality-unspecificindicator of semantic processing. This component was first reported in the visualmodality by Kutas and Hillyard  as anegStudies using the attentional blink paradigm  reportedIn sum, with respect to the automatic or controlled nature of semantic processingthere is contradictory evidence from the visual as well as from the acousticmodality with different indicators and paradigms. As discussed by Kiefer and Martens in termsKiefer and Martens  integratword pairs, there are to the best of our knowledge nostudies at sentence level. In contrast to the processing of word pairs, access tosemantic information during sentence reading is presumably more complex. Duringreading of single words lexical candidates have to be identified in the mentallexicon paradigm was applied (cf. Exp. 2). Sentence reading was combined with an additionaltask requiring the processing of non-linguistic material: a high or low tone had tobe classified according to pitch. The temporal overlap of sentence reading and theadditional task was manipulated by varying the stimulus onset asynchrony (SOA)between the tone and the target word, allowing for a precise control of thetime-course of interference. More specifically, in the paradigm used here, a tone(S1) and a visually presented target word (S2) - the adjective of a sentence - werepresented at one of three SOAs . The target word was always thepenultimate word in the experimental sentences (cf. Appendix for example sentences).Sentences required acceptability judgments immediately after sentence termination.Semantic processing was assessed by a sentence acceptability task based onappropriate and inappropriate Spanish noun-adjective pairings. Inappropriateadjectives were expected to induce N400 components in the ERP.From word level studies using PRP paradigms, different interference patterns areknown, such as a temporal delay of semantic processing, reflected in a shift of peaklatency of the N400 component  or N400 Recently, another ERP component, the P600, has been discussed to—at leastpartially—reflect semantic processing. The P600 is a long-lasting positivity,sometimes beginning as early as 200 ms after stimulus onset, reaching its maximumbetween 600 and 800 ms. Initially, the P600 has been attributed exclusively tosyntactic processing, because it was observed in the context of morphosyntacticviolations for both written and acoustic input in different languages . More reTo obtain baseline data and to test whether the Spanish sentences implemented in thePRP paradigm (see Exp. 2) would induce an N400 component, we first conducted asingle task experiment that included all the stimuli for the PRP experiment, butonly required acceptability judgments for the sentences; the additional tone was tobe ignored here. Experiment 1 served as a check whether the materials and procedurewould enable the recording of a N400 component and whether it would be modulated bythe different temporal overlaps (SOAs) between target word and tones when noresponse to the tone was required.Seventeen neurologically healthy, native Spanish-speaking university students(Faculty of Education) with normal or corrected-to-normal vision  took part in the experiment in return for coursecredits. All were right-handed , with avSEs = 1.97 vs. 1.94), accordingto the “Lexico Informatizado del Espańol” .The remaining fillers followed the structure of the experimental material,but verbs were transitive rather than intransitive and therefore required anobject complement . Half of the fillers were unacceptable sentenceswith semantic violations. Violations in the fillers occurred either in theverb or in the sentence’s final object. Filler sentences wereincluded because pre-tests without fillers had shown that the tone served asa cue to the upcoming violation, occurring directly after the tone andalways in the adjective of the experimental sentences. Hence participantsdid not have to deeply process the sentences and no N400 was induced. In thefillers the distance between tone and violation was more variable than inthe experimental sentences (more words per sentence or the violationoccurred in a different position and in a word category other than anadjective). In this way participants were forced to semantically process thesentences. All stimuli were presented in a white 20 pt font in the center ofa black computer screen at a viewing distance of about 65 cm, yieldingvisual angles between 0.7ş and 1.3ş in height and 1.1ş to6ş in width.TM software (BeriSoft Company).In addition to the visually displayed sentences, tones of 100 ms duration and300 or 600 Hz were presented. For experimental sentences the tone alwayspreceded the adjective at one of three possible SOAs. In case of fillersentences the tone occurred either before the subject, the predicate, or thesentence’s final object, being balanced over all filler sentences.Likewise, the SOA and, thus, the temporal overlap of tone and succeedingword was balanced. Manual responses to the sentences were recorded with twokeys operated by the index fingers; no response to the tones was required.Stimulus presentation and recording of responses were controlled byERTSThe basic structure of an experimental trial is illustrated in From the pool of 240 acceptable sentences and their 240 unacceptable versionstwo sets of 240 experimental sentences each were created, containing 120acceptable and 120 unacceptable versions. Within each set, no sentence wasrepeated and each sentence appeared only in one of its two versions(acceptable or unacceptable). SOA was randomly assigned to all the sentenceswithin a set, with an equal amount of acceptable and unacceptable sentenceswithin each SOA. Furthermore the assignments of high and low tones to eachsentence were counterbalanced. All responses had to be given within 2.5 safter the sentence-final word, which was always shown with a period. A newexperimental trial began either immediately after the response or when 2.7 shad elapsed since the offset of the final word.Participants were seated in a dimly lit, sound attenuated chamber. Bymanually pressing the left or right key participants made acceptabilityjudgments on each sentence. The response was to be given only after the endof the sentence. Participants were informed that they would hear high andlow tones, which were, however, irrelevant to their task. Furthermore, theyshould keep their eyes fixed to the center of the screen.Half of the participants responded to acceptable and unacceptable sentenceswith their right and left hand, respectively; for the others this assignmentwas reversed. The experiment consisted of 10 experimental blocks of 34trials each . Preceding the experimentproper 24 practice trials were repeatedly presented until the participantresponded correctly in all trials. Participants received visual feedbackabout the correctness of their response. After each block there was a shortbreak.TM). Scalp locations according to the revised version of the10/20 International System were Fp1, Fp2, F7, F3, Fz, F4, F8, FC3, FC4, T7, C3, Cz, C4, T8, TP7, CP3,CP4, TP8, P7, P3, Pz, P4, P8, PO7, PO8, O1, O2, plus a left mastoid (M1)electrode. All electrodes were initially referenced to the right mastoid(M2). Bipolar horizontal and vertical electrooculograms (EOG) were recordedfor artifact monitoring. Electrode impedances were kept below 5 k.ECITM electrode gel was used. The signals were recordedcontinuously with a band pass filter from 0.01 to 30 Hz and a sampling rateof 250 Hz.The electroencephalogram (EEG) was recorded by means of 27 tin electrodesmounted within an electrode cap , acceptability (2 levels), and electrode site (27levels) as independent variables. Error rates were subjected to ANOVAs, withrepeated measurements on SOA and acceptability. If appropriate, degrees offreedom were corrected according to Huynh and Feldt . Due to M = 20.4 vs. 15.0 %, SE = 1.89 vs.2.39), F = 5.65, p < .05. Forfactor SOA there was neither a significant main effect nor an interactionwith any of the other factors.Error rates were higher for acceptable than unacceptable sentences= 1.92, p = .005, however after Huynh and Feldt correctionthe effect failed significance. In the N400 time window the acceptabilityeffect was not significantly modulated by SOA. A post-hoc test at Cz and Pzelectrodes was performed, which yielded significant main effects of SOA,F = 10.5, p = .000, andacceptability, F = 18.7, p = .001,confirming the presence of an N400 at these typical sites. There was nointeraction of SOA and acceptability. Acceptability effects werestatistically confirmed also between 750 to 1050 ms , together with theinstructed priority of the tone discrimination task aimed at producing a PRPparadigm with variable SOAs . The SOA variation manipulated the temporaloverlap between the processes required by the choice response to the tone and theprocessing of semantically acceptable and unacceptable adjectives in thesentences.The experiment involved 20 neurologically healthy participants (16 women)with normal or corrected-to-normal vision; all of them were native Spanishspeaking university students (Faculty of Education) and received coursecredits for participation. Mean age was 20.05 years (range 18-26 years); 18participants were right-handed, two were left-handed . Ethical guidelineswere followed and participants signed an informed consent form.The material was mostly the same as in the single-task Experiment 1; twoacceptable adjectives were replaced because they had repeatedly causedincorrect responses. Mean values of cloze probability, number of letters,and word frequency of acceptable and unacceptable adjectives were notaffected by these changes.Apart from the additional task, the procedure was the same as in Experiment1. Accordingly, manual acceptability decisions had to be made to thevisually presented sentences; but now, also the tones were relevant,requiring choice responses. High- and low-pitched tones had to be respondedto with the left or right foot on keys, embedded in a footrest. The keyswere pressed with the big toes, shoes being taken off.The assignment of tone stimuli to response feet was counterbalanced acrossparticipants. Choice responses to the tones were to be executed withpriority over the sentence acceptability decisions. Temporal overlap (SOAs)of the tones with the critical words in the sentences was again varied inthree levels .As in Experiment 1 ten blocks of 34 trials were presented. Sixteen versionsof the experiment were generated balancing acceptable and unacceptableversions of the target words, mapping of hand and acceptability, andstimulus-response assignment of the tone task. High and low-pitched tonesoccurred equiprobably in each condition combination and independent of theacceptability condition. To familiarize participants with the dual taskrequirements, three practice blocks preceded the experiment proper, whichwere repeated until participants responded correctly. Visual feedback wasgiven during practice. Foot and hand responses were first practiced insingle task blocks and then in combination in one dual task block. Forpractice trials other language stimuli were used than in the mainexperiment.Data analysis of the behavioral and ERP responses to the visually presentedadjectives was conducted as in the single-task Experiment 1, with factorsSOA , acceptability(indicator of semantic processing), and electrode. Furthermore, alsoperformance (reaction times and error rates) in the additional tonediscrimination task was analyzed with repeated measures of SOA andacceptability.Ms = 4.18, 2.62, and2.93%, SEs = 1.10, 0.66, and 0.73, respectively),F = 3.22, p =.05. Tonediscrimination accuracy was neither affected by sentence acceptability asmain effect nor in interaction with SOA (Fs < 1).Reaction times in the tone discrimination task were neither significantlyaffected by SOA, F = 1.48  nor by sentence acceptability, nor by an interaction betweensentence acceptability and SOA .Responses in the tone discrimination task at SOA 100 were somewhat moreerror-prone than at SOAs 400 and 700 ,F = 8.69, p < .01. In contrast,to Experiment 1, there was also an effect of SOA: Error rates decreased fromshort to long SOA , F =16.56, p < .001. Furthermore SOA and acceptabilityinteracted, F = 4.74, p < .05 = 9.83, p <.01.Correctness of sentence acceptability judgments was significantly affected byacceptability. Acceptable sentences yielded fewer erroneous responses thanunacceptable ones  = .49, p> .05, but at SOA 400 and 700 this factor was significant . Acceptabilitydid not interact with factor Electrode although at SOA 700 there was a trendfor such an interaction (p = .09). In addition, an analysisincluding selected electrodes  was run. This yielded significant main effects ofSOA, F = 4.9, p = .016, andacceptability, F = 21.35, p = .000,as well as a strong trend for an interaction between these factors,F = 3.1, p = .057. Tests atsingle electrodes (Cz or Pz only) did not yield significant results,possibly because of the wide distribution of the N400.The patterns we observed in this experiment for the acceptability effectswere long-duration difference waves. Consequently, it was not feasible tomeasure peak latencies or peak amplitudes. Instead, the quantitativeanalysis was restricted to average amplitude measures between 400 and 600ms. As can be seen in p = .09). Excluding the factor electrode and runninganalyses only for the Pz electrode, where this component was largest, thisinteraction was significant, F = 3.96,p < .05. As in Experiment 1, effects ofacceptability were found in the later P600 time windows, between 1050 and1250 ms.Differences in amplitude between acceptable and unacceptable target nounswere again evident in the P600 time windows. This time, the P600 seemed tobe affected by SOA, with a reduction in amplitude (particularly at shortSOA) already in an earlier time window . On the ERP level,responses to the additional tone task at short SOA caused amplitude attenuationsof the N400 component to the critical words. Although the semantic task per sewas quite difficult, as indicated by high error rates in Experiment 1, it wasnot strongly postponed under conditions of high task overlap. In the followingthe pattern of results from both experiments shall be discussed in the contextof the controversy of automatic versus controlled semantic processing.The general aim of the present study was to investigate the effects of an additionaltask on indicators of semantic processing at sentence level. Main indicator ofsemantic processing was the N400 component of the ERP. According to the traditionalautomaticity view  semanticWhat we found in the present study were amplitude reductions that might be consideredas rather mild effects, in the sense that the N400 component was not drasticallydelayed. In contrast, in their dual task study with word pairs Hohlfeld et al. had repoAdditionally, in Experiments 1 and 2  we observeda P600 component. Its later segments (950 ms-1250 ms) were already modulated by theSOA in the single task of Experiment 1. In the dual task of Experiment 2, weobserved interference effects in both earlier (850 ms-950 ms) as well as later parts(1050 ms-1250 ms). The mild effects on the P600 observed in Experiment 2 mirror themild effects on the N400. Thus, we suggest that integrative processing as well assemantic processing were relatively stable during sentence processing.The observed patterns of results are difficult to explain in the context of atraditional understanding of automaticity but are in line with the attentionalsensitization model. We assume that the semantic processing pathway at sentencelevel is strengthened by continuously on-going priming from one word to the other.Kiefer and Brendel  demonstrThe PRP paradigm is usually employed to investigate whether a processing streamrequires a central process or depends on central attention. From interferenceeffects at high overlap between tasks (short SOA) the conclusion is drawn that theaffected process is a controlled, non-automatic process that requires centralattention or occupies a central processing bottleneck. As far as semantic processingis concerned interference effects in terms of temporal delays have been reported. Atword level Lien et al.  and HohlApart from latency delays also amplitude reductions of the N400 have been observed indual task studies . The obsThe amplitude reductions in the N400 in the present study are difficult to integrateinto a model that assumes a mere postponement of processing stages as suggested bythe bottleneck account cf. measuredThus, the amplitude reductions of the N400 observed in the present study could beexplained by such reconfigurations of the task set by SOA. Additionally, mechanismsare assumed that strengthen the semantic processing stream. To enhance performancethe cognitive system might be more flexible in dual task situations than previouslybelieved. Such a notion of flexibility is supported by findings from Lien et al., Exp. 4,Although we have to concede that the signal-to-noise ratio of the present studies wasnot optimal, the effects observed are in line with others and are theoreticallyplausible. Nevertheless further studies are needed to confirm and extend the presentfindings, which—to our knowledge—are the first at sentence level. Inaddition future research on semantic processing at sentence level might address thequestion, how the nature of the embedding sentences, for example their syntacticcomplexity or semantic constraints, will modulate the observed protective effect ina dual task situation.In sum, the findings from the present study contribute to the debate on the nature ofsemantic processing during reading and extend it to sentence level processing.Semantic processing was neither left completely intact nor strongly postponed by theadditional task. It is suggested that the observed amplitude reductions of the N400component were caused by a reconfiguration of the task set by the overlapping task,whereas the absence of postponement reflects a protective effect of the sentencecontext. This supports the idea of a flexible cognitive system that is able tostrengthen task-relevant pathways. The model of attentional sensitization hasintegrated this notion of flexibility and suggests that also automatic processes canbe modulated according to task demands. In this sense, the present findings indicatethat also semantic processing at sentence level is an automatic yet flexibleprocess.


The study aimed to provide estimates of the prevalence and psychosocial correlates of unintentional injury among school-going adolescents in Malaysia. Cross-sectional data from the Global School-Based Health Survey (GSHS) included 21,699 students (predominantly ≤13 to ≥17 years) that were selected by a two-stage cluster sample design to represent all secondary school students in Forms 1 to 5. The percentage of school children reporting one or more serious injuries in the past year was 34.9%, 42.1% of boys and 27.8% of girls. The two major causes of the most serious injury were “fall” (9.9%) and motor vehicle accident or being hit by a motor vehicle (5.4%), and the most frequent type of injury sustained was cut, puncture, or stab wound (6.2%) and a broken bone or dislocated joint (4.2%). In multivariable logistic regression analysis, sociodemographic factors , substance use (tobacco and cannabis use), frequent soft drink consumption, attending physical education classes three or more times a week, other risky behavior , psychological distress, and lack of parental or guardian bonding were associated with annual injury prevalence. Several factors were identified, which could be included in injury prevention promotion programs among secondary school children. Unintentional injuries are a major cause of disability and death among adolescents and youth, in particular in developing countries, including South Asia ,2. The uThe prevalence of one or more serious injuries in the past year among adolescents in Asian countries was 19.7% in China , 45.9% iFactors associated with unintentional injury among adolescents include: (1) sociodemographic factors, including male gender ,7,14,15 This study was a secondary analysis of existing data from the Global School-Based Health Survey (GSHS) from Malaysia. GSHS details and data can be accessed . The 2012 using international age- and gender-specific criteria [The study variables used from the GSHS  are desccriteria . The GSHcriteria .i.e., cluster sampling owing to the sampling of school classes. Associations between potential risk factors and injuries among school children, as largely identified from the literature review, were examined by calculating odds ratios (OR). Logistic regression was used for the examination of the impact of explanatory variables on the risk for injury (having had one or more serious injuries in the past 12 months = binary dependent variable). Potential multi-collinearity between variables was assessed with variance inflation factors, none of which exceeded the value of 1.6. Moreover, no statistical interaction (effect modification) was observed. Independent variables that significantly increased the injury risk in univariate analysis were included in the multivariable model. In reporting, weighted percentages are given, and the sample size reported reflects the sample that was asked the target question. The two-sided 95% confidence intervals are reported, and p-values less or equal to 5% are used to indicate statistical significance. The reported 95% confidence intervals and the p-values are both adjusted for the multi-stage stratified cluster sample design of the survey.Data analysis was conducted using STATA software version 12.0 . This software provides robust standard errors that account for the sampling design, The overall response rate, a combination of school and student response rates, was 88.7% . The finThe percentage of school children with one or more serious injuries in the past year was 34.9%, and that of those who had been injured more than once was 16.0%. The major cause of the most serious injury was “fall” (9.9%), followed by motor vehicle accident or being hit by a motor vehicle (5.4%), “something fell on me or hit me” (2.4%), and was attacked or abused or was fighting with someone (1.3%). Having been in a motor vehicle accident, falling and being attacked were significantly more frequent in boys than in girls. The most frequent type of injury sustained was “cut, puncture, or stab wound” (6.2%) and “a broken bone or dislocated joint” (4.2%) see .In multivariable logistic regression analysis, sociodemographic factors , substance use (tobacco and cannabis use), frequent soft drink consumption, attending physical education classes three or more times a week, other risk behavior , psychological distress, and lack of parental or guardian bonding were associated with annual injury prevalence. In terms of main specific types of injuries, older age (16 or 17 years or older), being male, substance use (tobacco and cannabis), frequent soft drink consumption, truancy, and psychological distress were associated with motor vehicle-related injuries. Furthermore, younger age, being male, tobacco use, frequent soft drink consumption, frequent attendance of physical education classes, and other risk behaviors (truancy and being bullied) were associated with fall injuries see .et al. [In this investigation of a large sample of secondary school-going adolescents in Malaysia, an annual injury prevalence of 34.9% was found, which is higher than in a few previous studies in Asian countries including China, Iran, Myanmar, and Taiwan ,6,8, simet al.  note thaIn agreement with previous studies ,7,14,15,Moreover, psychological distress was, in this study in multivariate analysis, significantly associated with injury. This finding was confirmed in previous studies ,18,19,20Although, in general, there were no age differences in relation to annual injury prevalence, there were age differences regarding specific types of causes of injuries; older age (16 or 17 years or older) was associated with a motor vehicle-related injury and younger students had a higher annual prevalence of falls than older students. The study found that attending physical education classes three or more times a week was associated with overall injury and fall injuries, in particular. Previous studies found that frequent participation in sport activities  was assoThe study found that other risk behaviors  were associated with injury, which is partly confirmed in other studies ,17,18. SOverall, the study found, as in some previous studies ,17, thatvs. those sustained in the home as well as injuries that were inflicted by others vs. those that were self-inflicted were not assessed in the Malaysian GSHS. Depending on, for example, the location of where most injuries had occurred, such as at school, at home, or on the road, interventions could be more specifically targeted [This analysis represents one of the first national examinations of adolescent injury in Malaysia. Injuries sustained outside the home targeted .The study focused on secondary school-going adolescents who may not be representative of all adolescents in Malaysia as the prevalence of injury events and associated factors may be different between school-going and non-school-going adolescents. Further, the questionnaire used was completed by self-report, which may have introduced a reporting bias. Moreover, this study was based on data collected in a cross-sectional survey, and we can therefore not ascribe causality to any of the associated factors in the study. The analysis was limited to the variables which had been included in the GSHS in Malaysia, and other additional risk factors and the location of the injury could have been included . AnotherIn this study, a high annual prevalence of injury was found among a large sample of school-going adolescents in Malaysia. Various associations with injury were identified, including psychological distresses, substance use, truancy, being bullied, ever having had sex, and frequent soft drink consumption, which may be considered in an integrated approach to injury prevention and safety promotion strategies among school children.


Background. Prevalence of childhood asthma varies significantly among regions, while its reasons are not clear yet with only a few studies reporting relevant causes for this variation. Objective. To investigate the potential role of city-average levels of air pollutants and climatic factors in order to distinguish differences in asthma prevalence in China and explain their reasons. Methods. Data pertaining to 10,777 asthmatic patients were obtained from the third nationwide survey of childhood asthma in China's urban areas. Annual mean concentrations of air pollutants and other climatic factors were obtained for the same period from several government departments. Data analysis was implemented with descriptive statistics, Pearson correlation coefficient, and multiple regression analysis. Results. Pearson correlation analysis showed that the situation of childhood asthma was strongly linked with SO2, relative humidity, and hours of sunshine (p < 0.05). Multiple regression analysis indicated that, among the predictor variables in the final step, SO2 was found to be the most powerful predictor variable amongst all . Furthermore, results had shown that hours of sunshine  was a significant component summary predictor variable. Conclusion. The findings of this study do not suggest that air pollutants or climate, at least in terms of children, plays a major role in explaining regional differences in asthma prevalence in China. Asthma is one of the most chronic inflammatory disorders characterized by recurrent attacks of wheezing, breathlessness, cough, and/or chest tightness, which vary in severity and frequency from person to person. The prevalence of allergic airway diseases, such as asthma and rhinitis, has been increasingly common to epidemic proportions worldwide. About 334 million people worldwide are now suffering from asthma . And ChiAnd, more importantly, there are significantly striking differences in asthma symptoms worldwide , 4. The A large number of studies have confirmed that respiratory diseases are related to the physical characteristics of the living area: asthma has multifactorial etiology, including exogenous factors like air pollution and climate. The link between urban air pollution, climatic factors, and asthma is stably established on the individual level. However, there are merely a few analyses developing a comparative approach of differences in asthma morbidity among places, such as cities. In addition, levels of outdoor air pollutants have been associated with asthma incidence but not clearly with asthma prevalence at the population level . Under tThe data of asthma, air pollutants, and other climatic factors were obtained from published journals or government departments. Asthma definition in this survey was based on the Global Initiative for Asthma, Chinese Guideline of Pediatric Asthma published in 2008 . In the Asthma data in this study were collected from the open-access Chinese Journal of Pediatrics . The ChiApproval for the use of the data was obtained from the National Cooperative Group on Children Asthma. And informed consent was obtained from the next of kin, caretakers, or guardians on behalf of children. The consent on behalf of the children enrolled was written. The study design was approved by the ethics committee of the National Cooperative Group on Children Asthma.µm PM10 (µg/m3), sulfur dioxide SO2 (µg/m3), and nitrogen dioxide NO2 (µg/m3). All the data were obtained from the State Statistical Bureau Network which collected annual average concentrations of air pollutants through automated fixed-site monitoring stations. The data were regularly recorded from January 1, 2009, to December 31, 2010. We collected and calculated the 2-year average (from 2009 to 2010) values of these variables, yet numbers of air quality monitoring stations in 43 cities were not available.Air pollution data included the annual (from 2009 to 2010) average values for particulate matter less than or equal to 10 2 and for 24-hour mean concentrations of SO2 were 20 µg/m3, 40 µg/m3, and 20 µg/m3, respectively [The World Health Organization Air Quality Guidelines for annual mean concentrations of PM10 and NOMeteorological data included the annual average values for temperatures (°C), relative humidity (%), precipitation (mm), and hours of sunshine (h). Obtained from the State Statistical Bureau Network and the China Meteorological Administration, these data were regularly recorded from January 1, 2009, to December 31, 2011, as well. We collected and calculated the 2-year averaged (from 2009 to 2010) value of these variables. The numbers of weather monitoring stations in 43 cities were also not available.2, and NO2) and climatic factors . Statistical significance was set at p < 0.05, and all statistical analyses were performed with SPSS software version 20.0.Descriptive statistics were used to summarize the general characteristics of the data included in this study. The Shapiro-Wilk test revealed a normal distribution of tested variables. The mean and standard deviation (SD) of each of the variables were then calculated. Associations were assessed with Pearson correlation coefficient and multiple regression analysis. Prevalence of asthma was the dependent variable, and independent variables were air pollutants  than in school-age children  and infants . Within the total survey cohort, 12,997 (3.00%) patients of Han descent, 87 (2.73%) patients of Manchu descent, and 157 (2.39%) patients of Hui descent were asthmatic. In the asthmatic patients, respiratory tract infection  and changes in the weather conditions  were the most frequent triggers of asthma exacerbation. Common clinical manifestations of asthma were cough  and wheezing . A family history of allergies was reported in 45.2% of patients , and a personal history of allergies was reported in 72.5% of patients . Regarding the frequency of asthma attacks each year, 87.7% of patients reported 1–5 asthma attacks, 10.1% reported 6–10 attacks, and only 2.2% reported more than 10 attacks. To treat asthmatic children, bronchodilators were used in 71.4% of cases , inhaled corticosteroids in 58.7% , and antibiotics in 75.1% .The national prevalence of asthma in male and female children was 3.51% and 2.29%, respectively. The prevalence of asthma was significantly higher in preschool children , together with the annual mean levels of climatic factors  for each of the 43 cities. There were statistically significant differences in asthma prevalence, air pollutants, and climatic factors between cities , relative humidity , and hours of sunshine . As can be seen, these correlations between PM10, NO2, air temperature, precipitation, and asthma prevalence were not statistically significant. Therefore, the multiple linear regression analysis with a dependent variable of asthma prevalence and independent variables of SO2, relative humidity, and hours of sunshine was used to predict asthma prevalence. The prediction model was statistically significant  and accounted for approximately 30.3% of the variance of asthma prevalence . Multiple regression analysis indicated that, among the predictor variables, SO2 was found to be the most powerful predictor variable amongst all . Furthermore, results had shown that hours of sunshine  was a significant component summary predictor variable. However, relative humidity was not significantly predicted to be the outcome measure.The results of the Pearson correlation coefficient and multiple regression analysis describing the relationship between asthma prevalence and risk factors (air pollutants and climatic factors) are shown in Tables 2 and hours of sunshine. Before the discussion of the findings, it is important to address methodological aspects of this study. This paper does not focus on the individuals themselves but cities, which is in contrast to many epidemiological study designs where information is available at the individual level. In addition, our data was available at annual mean level rather than daily. The size of the study was large, and although large-scale researches were accomplished by ISAAC in reference to asthma for air pollution [The aim of this study was to investigate the potential role of city-average air-pollutant levels and climatic factors in explaining regional differences in asthma prevalence in China. Variables for significant correlation coefficients were also obtained in the multiple regression analysis. They were the mean SOollution , 11 and ollution , 12, theA growing body of research on time trends is devoted to the relationship between allergic respiratory diseases including asthma and environmental factors, climate conditions and air pollutants, but results on effects of these variables on asthma are still unclear and current knowledge is provided by epidemiological and experimental studies.2 no longer provides a statistical explanation for variations in asthma prevalence across urban units. Interestingly, the relationship between childhood asthma and SO2 was consistently negative. And surprisingly, these annual mean levels of PM10 (0.091 mg/m3), NO2 (0.042 mg/m3), and SO2 (0.042 mg/m3) detected during the sampling period did not conform with the data from World Health Organization Air Quality Guidelines [The adverse effect of air pollution on respiratory health has been well established in the literature , 14, butidelines –18, inclidelines , 15, 16 idelines . The disThe influence of short-term climate change on asthma exacerbation is well established, but its long-term influence on disease occurrence is little studied.Regarding the effect of climatic variables, a large number of studies have confirmed that temperature is related to asthma , 20–31. In the present study, annual mean relative humidity was not related to asthma prevalence. Indeed, relative humidity has been associated with the prevalence of wheezing , 29, symThese few studies analyzing associations of asthma with precipitation , 31, 36 The reasons for the inconsistencies among our study and previous studies are possibly regional discrepancies such as geographical environment, economic development, and density of the local populations. Additionally, study design, analyzing method, subgroups of children, and different parameters may also contribute to the controversies. Thus, a direct comparison of results is not accurate.On the other hand, there are still several limitations in this study. First, modeled estimates of air pollutants and climatic factors at city level are imprecise and incomplete in terms of personal exposure to ambient air pollutants. Second, collinearity between variables and the time series impeded the evaluation of particular pollutants or climatic factors that had a direct adverse effect on the prevalence of asthma. Third, other factors such as the indoor environment , economiTo conclude, our study showed associations between long-term exposure to urban air pollution and climate conditions, using Pearson correlation coefficient and multiple regression analysis capturing variations within communities, and asthma. First, the study shows the surprising results for the contribution of pollution factors. In contrast to reports from within-community studies of individuals exposed to traffic pollution, we found no evidence of a positive relation between air pollution and asthma prevalence. Meanwhile, our results suggest that climate may affect the prevalence of asthma, yet climate does not play a major role, at least in terms of children.


AbstractSciaenidae (30 spp.), Carangidae (26), Haemulidae (24), Serranidae (21), Paralichthyidae, and Gobiidae (13). Of the total species list, 134 represent first records for the Michoacán State, and one is a first record for Mexico. The results expand the number of known fish species of the Michoacán coast by almost one third and will help to develop conservation and management plans for this coastal zone.This study is the first to complete an intensive and comprehensive list of the ichthyofauna of nearly all ecosystems of the Michoacán coast, Mexico. The resulting systematic checklist, supplemented with information from the literature and scientific collections, comprises 440 species belonging to two classes, 31 orders, 104 families, and 264 genera. The families with the highest number of species were  Mexico has a wealth of both marine species and ecosystems. The country has the world’s twelfth longest marine territory, including both Atlantic and Pacific oceans. The geographic and geological history, as well as the ecological and biological richness of areas such as the Gulf of California, the Oceanic islands, and the Caribbean, expands the diversity of Mexican marine life , includiTropical Eastern Pacific (TEP), which comprises three provinces: Cortez or Sinuscaliforniana, Mexican, and Panamic . The TEP could be considered an area of low richness compared to other biogeographical regions of tropical seas, such as the Indo-Malaysian or the Great Caribbean. However, approximately 71% of identified TEP fish species are considered endemic, making it the tropical region with the highest rate of endemism per unit area in the world  represent, on the other hand, 10% of the total.The Mexican tropical Pacific is part of the biogeographic region known as the ic sensu. An estihe world . AccordiThe Mexican province is highly productive due to the convergence of the Costa Rica Coastal Current and the California Current, favoring the presence of tropical, temperate, and transitional fish species . Based oPageBreakdiffering markedly in the marine ecosystems and consequently in the fish species present: (1) the municipalities of Lazaro Cardenas and Coahuayana, which are characterized by coastal plains, with a wide sandy coastline, mangroves, and estuary zones, (2) and the municipality of Aquila comprising numerous cliffs extending into the sea and forming wide zones of rocky reefs, coralline patches, and intertidal pools; estuaries in the last zone are scarce and differ in size and dynamism from those found in Lazaro Cardenas and Coahuyana , the Colección Nacional de Peces (UNAM), the Instituto de Ciencias del Mar y Limnología Colección Ictiológica del (UNAM) and the Marine Vertebrate Collection, Scripps Institute of Oceanography (SIO). In addition, records from the data base of the fish collection of the California Academy of Sciences(CAS) were reviewed. These investigation also included an extensive review of specialized publications  and reports of specimens deposited in ichthyological collections recognized by the Secretaria de Medio Ambiente y Recursos Naturales, México, or specimens of which identification was corroborated by experts.The study area encompassed the coastline of Michoacán state, with 110 locations directly sampled .Most specimens were photographed upon collection; tissue samples were taken and deposited at the tissue collection of the obatidae , Gerreiderreidae , Atherinnopsidae , Labrisoisomidae , Blenniienniidae , Chaenopnopsidae , Tripterrygiidae , and forål, 1775 , Tomicodde, 1800 , and Albli, 1777 . All speThe systematic arrangement followed Finally, a zoogeographical affinity analysis of the species, based on the biogeographical regionalization proposed by Sampling was performed in 13 intertidal pool sites, 20 rocky reef sites, three coralline communities, two artificial reefs, 22 estuarine ecosystems, and 50 soft bottom and open sea sites, collecting 6963 fishes.Sciaenidae (30), Carangidae (26), Haemulidae (24), Serranidae (21), and Paralichthyidae and Gobiidae (13). The genera with the highest number of species were Lutjanus Bloch, 1790 (9), Carcharhinus Blainville, 1816 (7), Anchoa Jordan and Evermann, 1927 (6), Diplectrum Holbrook, 1855 (6), and Caranx Lacepède, 1801 (5).The compiled systematic list of ichthyofauna of the Michoacán coast comprises 436 species belonging to two classes, 31 orders, 104 families, and 260 genera , Aetobatuslaticeps , Apogonpacificus , Coryphaenahippurus Linnaeus, 1758, Kyphosusocyurus , and Novaculichthystaeniourus . Of the 436 species, 131 were new records for the Michoacán State, and Canthigasterjanthinoptera  was a new record for Mexico  were collected in reefs, 57 (19%) in the pelagic-demersal zone, 46 (15%) in estuarine zones, 17 (6%) in the demersal zones, and 14 (5%) in rocky intertidal zones. Forty-seven species were collected in more than one habitat type (Table Carangidae (17), Haemulidae (15), Sciaenidae (13), Serranidae (10), and Lutjanidae (7). Of these, 104 (68%) were commercially valuable, 23 (15%) were used for direct consumption or as bait. Twenty-seven (18%) were bycatch, that are normally rejected and thrown back  were widely distributed from the San Diegan province to the Panamic province, and 3 (1%) from the San Diegan province to the Mexican province. The largest number of species, 216 (49.2%), were found in the three provinces of the TEP; 15 PageBreak(3%) were restricted to the Cortez and the Mexican provinces; 35 (8%) were limited to the Mexican and Panamic provinces; and 2 (0.5%) were endemic to the Mexican province , Sicydiummultipunctatum Regan, 1905, Gobiesoxmexicanus Briggs & Miller, 1960, and Poeciliabutleri Jordan, 1889 (fresh water). The introduced species Pterygoplichthysdisjuntivus  and Cyprinuscarpio Linnaeus, 1758 were also omitted.Forty-six (11%) of the species were circumtropical, 27 (6%) transpacific, and seven (2%) amphi-American; whereas 350 (81%) belonged to the ies, 216 9.2%, werPageBreakzones with increased sampling effort  were collected from Barco Hundido del Faro de Bucerias , which identification was corroborated by BLAST (http://www.ncbi.nlm.nih.gov/genbank) and by boldsystems (http://www.boldsystems.org), searches showing 99% similarity in the cytochrome oxidase subunit 1 gene (COX-1) to specimens identified as Canthigasterjanthinoptera from the south Tropical Eastern Pacific and the Indo-Pacific Ocean (GenBank accession numbers: KX505745 and KX505746). One specimen of Bathygobiusandrei  (Gobiidae) was also reported in the Chuta estuary , representing an extension of its previously known northern distribution limit of the coast of Chiapas  was collected from rocky reef in Faro de Bucerias location (18°20'50"N and 103°30'37"W), extending its extension range in the TEP.The present checklist represents the most updated and comprehensive systematic list of fishes recorded from the coast of the Mexican State of Michoacán. Of the species cataloged, 30% were first records for this State. The highest proportion of cataloged species was collected in reefs (40%). The pelagic zone accounted for 24% of the species collected, indicated a requirement for future studies of the demersal and pelagic ort Fig. . This coort Fig. . Anotherort Fig. , for whi Chiapas . One speCarangidae ), Lutjanidae ), Paralichthyidae ), Centropomidae , and Serranidae ) (Table Sphyrna sp.), were captured, as well as pregnant Rhinobatosglaucostigma Jordan & Gilbert, 1883, Gymnuramarmorata , and Urotrygon spp. In general, the elasmobranchs, due to their unique biological and ecological characteristics, present low population growth and are considered highly vulnerable , LutScorpaenamystes  reaches 35.6 cm and is marketed in regions such as Baja California. Trichiurusnitens Garman, 1899 supports a small fishery in the central portion of the littoral zone of Ecuador , whereas, in Michoacán, this species is not used for human consumption.Bycatch in commercial fishing is frequently used for personal consumption, bait (~50%), or discarded Fig. . SpeciesPageBreakTEP provinces, with 8% of the species reported only in the Panamic and Mexican provinces, being mostly of tropical affinity, agreeing with previous fish fauna surveys in the area (It was observed that 49% of the listed species are reported as also occurring throughout the three Although visual censuses and photo identification of fish species is widely used for the study of richness, diversity, and ecology of marine habitats (Aguilar-Palomino 2002, CPUM collection by 19%, since the majority of marine species previously collected remain in collections outside of Michoacán. Many species reported in this study as new records (Table The collections obtained in the present study enriched the records of the


Post-acute community-based rehabilitation is effective in reducing disability. However, while social participation and quality of life are valued as distal outcomes of neurorehabilitation, it is often not possible to observe improvements on these outcomes within the limited time-frames used in most investigations of rehabilitation. The aim of the current study was to examine differences in the sequence of attainments for people with acquired brain injury (ABI) undergoing longer term post-acute neurorehabilitation. Participants with ABI who were referred to comprehensive home and community-based neurorehabilitation were assessed at induction to service, at 6 months and again at 1.5 years while still in service on the Mayo-Portland Adaptability Index (MPAI-4), Community Integration Questionnaire, Hospital Anxiety and Depression Scale, and World Health Organisation Quality of Life measure. At 6 months post-induction to service, significant differences were evident in MPAI abilities, adjustment, and total neurodisability; and in anxiety and depression. By contrast, there was no significant effect at 6 months on more socially oriented features of experience namely quality of life (QoL), Community Integration and Participation. Eighteen month follow-up showed continuation of the significant positive effects with the addition of QoL-related to physical health, Psychological health, Social aspects of QoL and Participation at this later time point. Regression analyses demonstrated that change in QoL and Participation were dependent upon prior changes in aspects of neurodisability. Age, severity or type of brain injury did not significantly affect outcome. Results suggest that different constructs may respond to neurorehabilitation at different time points in a dose effect manner, and that change in social aspects of experience may be dependent upon the specific nature of prior neurorehabilitation attainments. Acquired brain injury (ABI) is the leading cause of death and disability in young people aged 18–24 years: it also disproportionately affects children (up to 4 years-old) and people aged over 65 years . ABIs caThe value of specialized rehabilitation for brain injury, including community-based rehabilitation, is becoming increasingly apparent in terms of both functional outcomes , and cosA number of features of participant’s experience have been suggested to be important targets for, and outcomes of, intervention. Changes in neurodisability is one of the most prominent outcomes and in addition to targeting changes in physical and cognitive abilities and mental health, rehabilitation programs increasingly utilize more socially moderated factors such as quality of life and participation in society . Such soMental health difficulties, particularly anxiety and depression have relatively common currency in ABI and pose a significant barrier to rehabilitation progress . Indeed Community participation and quality of life (QoL) are increasingly valued as neurorehabilitation outcomes . ParticiIn terms of QoL, There is some recent evidence supporting a dose-effect relationship on outcome of neurorehabilitation. In a large study of a cohort of people with CVA , particiThe objectives of the current study were therefore, firstly to examine whether participants demonstrate significant improvement on a broad number of domains assessed at shorter and longer durations of community-based neurorehabilitation: namely neurodisability, community integration, mental health, and quality of life. Secondly to examine the contribution of differences in clinical features of injury such as age at onset, injury severity, type and duration of brain injury, age, and sex on changes from pre-treatment to follow-up. Thirdly to investigate whether changes in the more socially oriented factors including QoL, participation and social/community integration occurred later than changes in neurodisability and mental health outcomes and furthermore whether such changes in QoL, participation and community integration may be dependent upon prior changes in neurodisability and mental health outcomes.U = 118.5, p = 0.047), for parsimony, these six participants were excluded from further analysis. The final cohort consisted of 77 participants who were assessed at induction to service (time 1), 6 months later (time 2) and at 1.5 years post-induction (time 3). In addition to the cohort of participants followed up over three time points during their service, a total of 151 additional people with ABI who were referred to service after the cohort had been established, during this 2 years period were assessed at induction to the service for comparison purposes with all induction data of the cohort. The purpose of this cross sectional sample was to examine goodness of fit of the cohort to those routinely referred to post-acute neurorehabilitation services.Eighty three people were eligible for participation in the cohort study. A total of six participants dropped out of the study between their initial induction assessment and follow-up assessment. Given that there was only one significant difference between the induction data for these six participants and the cohort .The protocol was approved by the ABI Ireland national research ethics committee and all participants provided full consent for participation. Assessments were administered by staff as semi structured interviews. All clients of the service were eligible for participation where the inclusion criteria was deemed appropriate by members of the Clinical Service Team.Table 1).Demographic, clinical and social information was recorded at induction to the study  scores, duration of Post-Traumatic Amnesia, and Loss of Consciousness (LOC) using the following procedure: Severe Brain Injury = GCS score less than 9, LOC longer than 24 h, or PTA longer than 1 week. Moderate Brain Injury = GCS score of 9–12, LOC of 30 min to 24 h, or PTA of 24 h to 1 week. Mild Brain Injury = GCS score higher than 12, LOC less than 30 min, or PTA less than 24 h. If more than one indicator was present and differed in level of severity, the more severe level was assigned. For other causes , severity was assessed by multidisciplinary team discharge report from acute  rehabilitation services specifying moderate and severe disability.Participants completed the following measures of Neurodisability, Community Integration, Mental Health, and Quality of Life at induction to post-acute neurorehabilitation, at 6 months follow-up and at 1.5 years post-induction.The MPAI-4 is a widely used measure of limitations imposed by brain injury . The meaThe Community Integration Questionnaire CIQ; , is a brThe Hospital Anxiety and Depression Scale HADS; , a 14-itThe WHOQoL-BREF is a 26 item international cross-culturally comparable quality of life assessment instrument. The assessment examines a person’s Quality of Life in relation to four domains: QoL related to Physical Health, Psychological Health, Social Relationships, and the person’s living Environment. Higher scores denote better QoL. The measure has demonstrated appropriate reliability and validity , and hasZ’s > 1.21, p’s > 0.14), means and standard deviations were calculated for the four main outcome measures and subscales as appropriate. Repeated measures analysis of variance models were used to model the means of each of the outcome measures over time. Effect sizes [Partial eta squared (Post hoc pairwise comparisons were conducted and a Bonferroni adjustment (α = 0.004) was performed to examine differences between outcomes at time 1 and time 2, and between time 2 and time 3. Categorical data were analyzed by Chi square test and longitudinal categorical data by Cochrane’s Q-test for three time points and the McNemar test for two time points. Repeated measures ANCOVA was used to examine differences in outcome for the two principal causes of ABI; injury resulting from an external force (TBI), or injury resulting from an internal disease process , and severity of injury. Zero order correlations and multiple regression analysis were used to examine the potential influence of prior changes in neurodisability and mental health on subsequent changes in QoL and Participation.Descriptive statistics were computed for variables relating to injury and demographics. Q-Q plots and Kolmogorov–Smirnov test were used to examine the distribution of outcome data. Given that outcome data showed no significant deviation from normality  and people who were referred for service in the 2 years of the study (N = 158) on any of the outcome assessments at induction . There were also no significant differences between the cohort and single assessment groups on participants’ age, age at onset of their ABI, or duration with ABI , or on clinical severity of their injury . There was also no significant difference between the cohort and people who completed their assessments at induction on numbers of people with CVA vs. TBI . The cohort was therefore not unrepresentative of the profile of people with ABI routinely referred to post-acute neurorehabilitation services in the Republic of Ireland.There were no significant differences between the cohort , participants’ age, duration with ABI, or age at onset of their injury .At induction to the study, cohort participants’ performance on neurodisability, community integration, distress and QoL did not differ as a function of gender (t-tests (α < 0.004) demonstrated that when compared to participants with mild/moderate injury, people who had sustained a severe brain injury were functioning at a poorer level at induction in terms of the MPAI assessment of Abilities , Adjustment , and Participation . Whether a person sustained a moderate or severe brain injury did not significantly affect scores on Community Integration, Mental Health or QoL at induction . Means and standard deviations for all outcome measures are presented in Table 2.Independent F1,76 = 15.29, p = 0.001, F1,76 = 36.87, p = 0.001, F1,76 = 19.33, p = 0.001, F1,76 = 33.82, p = 0.001, A single repeated measures analysis of variance was used to model the MPAI Abilities, Adjustment, Participation, and Total scale score over the three time points. Significant increases were apparent for people with ABI in terms of their Abilities , Adjustment , and total Neurodisability , but not for Participation .Pairwise comparisons revealed significant induction to 6 months follow-up improvements for Abilities , Participation , and total Neurodisability . However, changes in Abilities failed to reach significance .Significant pairwise comparisons of 6 months to 1.5 years data were found for Adjustment .Using published MCID scores , for theF1,75 = 6.82, p = 0.001, t76 = 2.78, p = 0.001), however, the 6 months to 1.5 years data was not significant .Repeated measures ANOVA demonstrated that mean reductions in depression scores over time were statistically significant (n = 30/77) scored above the cut-off for clinically relevant depressive symptoms . This figure had fallen to 24.6% (n = 19/77) after 6 months of rehabilitative intervention, and to 20% (n = 15), 1 year later (1.5 years post-baseline). This represented a significant effect . Post hoc McNemar test with Bonferroni correction suggested that the principal difference was between induction and 6 months follow-up only (p = 0.01).Using the established cut-offs for the presence of clinical distress, at induction to the study 39% of clients  but not 6 months to 1.5 years . Mean (SD) scores are presented in Table 2.Repeated measures ANOVA suggested that mean differences in anxiety over time were statistically significant (n = 24/77), scored above the cut-off for clinically relevant symptoms of anxiety. This figure had fallen to 14.3% (n = 11/77) after 6 months of rehabilitative intervention, and had increased slightly to 15.6% (n = 12/77) 1 year later. Cochrane’s Q-test suggested this represented a significant effect , with post hoc McNemar tests again finding that the significant reduction was between induction and 6 months only (p = 0.001).In terms of clinically relevant anxiety , at induction 31.2% of clients assessed . Comparisons of 6 months to 1.5 years follow-up revealed significant improvements on QoL related to Physical Health , Psychological health , but not Social aspects of QoL .Pairwise comparisons (α < 0.004) demonstrated no significant induction to 6 months follow-up improvements for any of the quality of life measures (We next examined whether the significant changes in QoL and Participation from 6 months to 1.5 years were dependent upon prior (induction to 6 months) changes in neurodisability and mental health.Table 3) which suggested that significant 6 months to 1.5 years changes in Participation was related to prior improvements in the Neurodisability factors of Abilities  and Adjustment  and in prior changes in Depression . t-test showed no significant effect of injury severity on 6 months to 1.5 years Participation scores .Prior to building the regression model, correlation analysis was undertaken (Table 4) for change in Participation was significant for the three variables – prior improvements in Depression, Adjustment to disability and Abilities . However, only prior improvement in Abilities predicted subsequent improvements in Participation in the final regression equation .The regression model  and Adjustment . Neither changes in QoL related to psychological health nor social aspects of QoL were associated with prior changes in neurodisability or mental health (r’s < 0.17). The regression model for positive change in QoL-related Physical health at time 3 was initially significant for prior improvements in both Abilities and Adjustment . However, improvements in QoL-related Physical Health was solely predicted by prior improvements in MPAI Adjustment only  in the final equations (Table 4).The significant improvement in QoL related to Physical Health at 1.5 years follow-up was associated with prior significant changes from baseline in Abilities , the participants in this study were individuals at the more severe end of the brain injury spectrum and as such results may not generalize to individuals with less severe brain injuries or with a shorter duration of injury.Nonetheless, this study has demonstrated improvements over time for participants in long-term service with moderate to severe brain injury across a range of measures of outcome. Results also suggested the presence of a dose effect which varied as a function of the nature of the outcome, with some outcomes showing the need for increased time duration, and others demonstrating consolidation after a shorter duration of intervention. Importantly, this study revealed that significant changes in more nuanced person-centered and social aspects of outcome such as quality of life and participation only occurred after significant changes in aspects of neurodisability had become established. These results have obvious implications for specifying the sequence of neurorehabilitation interventions in order to best optimize proximal and distal treatment outcomes, and relates, as has been proposed previously e.g., , to the 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.


The aim of the study was to explore the association between mucin 5ac expression and cancer prognosis.A systematically comprehensive search was performed through PubMed, the Web of Science, and the China National Knowledge Infrastructure (CNKI). The prognostic value of mucin 5ac expression in cancer patients was evaluated.P = 0.003). This association was also detected in a biliary subgroup  and a gastrointestinal subgroup . In the geography subgroup analysis, a statistical association was found in the Asian subgroup . In the clinical characteristics analysis, a statistical association was found between the hyper expression of mucin 5ac and lymphatic metastasis.The overexpression of mucin 5ac was found to be significantly associated with a poor prognosis in cancer patients (pooled HR: 1.53, 95%CI: 1.158–2.028, We indicated that mucin 5ac is a promising prognostic predictor for cancer, especially for biliary and gastrointestinal cancer, and is more suitable for predicting cancer prognoses in Asians. And no ethical approval was needed because our meta-analyses were based on data from previously published studies.A computerized literature search was conducted of PubMed, the Web of Science, and the China National Knowledge Infrastructure (CNKI) with the following strategy:  from 2000 to March 2015 in order to identify all studies that explored the association between mucin 5ac levels and cancer prognoses. Meanwhile, the time period, sample size, population, types of clinical trials, or types of reports on the retrieved studies were not limited, and we only reviewed articles in the English and Chinese languages. Every retrieved study was inspected manually for the inclusion criteria. To explore additional studies, we also examined the references of the included articles and reviews. The last search was carried out in May 2015.Publications were included in our analysis if they included a 1) proven diagnosis of cancer in humans, (2) an evaluation of the association between MUC5AC and cancer prognosis , (3) mucin 5ac quantity (protein or mRNA) analysis of the primary tumor (not in metastatic tumor or in tumor adjacent tissues) or in the serum, or an (4) eligible hazard ratio (HR) with 95% confidence intervals (CI) or other available data for estimating the HR with a 95%CI (including extracting data from K-M curves). We excluded non-RCTs, case series or case reports, review articles, editorials, letters, and comments. We also excluded non-English studies, nonhuman experiments, and studies with insufficient data. For overlapping studies, we took the largest study or the study with the first published samples. The study selection flow diagram is shown in Figure  proven d16 For studies that only provided K-M survival curves, we extracted the necessary data. In order to extract more robust data from the K-M curves, 2 methods were used 21 .21 We also collected the following data from each study: first author's name, year of publication, quantitative method, cancer type, sample size, country, cut-off value, specimen, and follow-up duration. Geography was categorized, based on the source country, as “Asian” or “non-Asian” (presented in Table Two independent researchers (XW and XH) extracted data according to the inclusion criteria, and discordant studies were submitted to a third investigator (BR) for further review. To avoid overlapping patient populations, we compared data sources and geographic locations. The hazard ratios with the corresponding 95% confidence interval were extracted from every included study. For studies providing HRs, we obtained data directly.A quality assessment of the included studies was evaluated with the Newcastle–Ottawa quality assessment scale (NOS) ranging from 0 to 8 by 2 independent investigators (XW and FY). Studies with an NOS score ≥6 were considered high-quality studies. Studies from conference abstracts were defined as low-quality studies. Any inconsistencies were resolved by joint discussion.P values and I2 values. If the P value was <0.05, indicating the presence of heterogeneity in studies, a random-effects model (based on the DerSimonian and Laird method) was used to assess the HRs and corresponding 95% CIs. Otherwise, the fixed-effects model (based on the Mantel–Haenszel method) was applied.25 An observed HR >1 indicated a worse prognosis in patients with mucin 5ac high expression and an HR <1 suggested a better prognosis. Thereafter, we performed subgroup analyses to explore sources of heterogeneity. A sensitivity analysis was performed by the sequential omission of individual studies. Publication bias was evaluated using a funnel plot and Egger's regression asymmetry test. Meanwhile, pooled odds ratios (ORs) with 95% CIs were calculated to describe the association between mucin 5ac expression and clinicopathological parameters. All statistical analyses were performed with STATA software , and a P-value <0.05 was considered significant.HRs with 95% CIs were calculated to evaluate the relationships between mucin 5ac levels and overall survival. The heterogeneity among studies was assessed by calculating relevant 26 26 were included according to inclusion criteria. Imai and Shiratsu's studies included 2 different survival analyses separately . Thus, a total of 17 studies, 2102 patients, were analyzed in this meta-analysis. All of the patients were pathologically diagnosed. Of all the eligible studies, 13 were conducted in Asian, whereas 4 in non-Asians areas, 9 were gastrointestinal cancers, 4 were biliary cancers, and 4 were other cancer types . Details are shown in Table The study selection processes using electronic database were presented in Figure P = 0.003, heterogeneity, P = 0.012, Figure A total of 2102 patients from 14 articles were enrolled in this analysis  and gastrointestinal carcinoma , which was the predominant method used to detect mucin 5ac expression (see details in Table P = 0.01).As the P = 0.385 . No evidence of asymmetry was found in our funnel plot . This indicates that mucin 5ac can be used for predicting cancer prognoses. In order to refine a more detailed conclusion, we stratified the analysis of enrolled studies. First, we performed a subgroup analysis by cancer type. Four studies were enrolled in the biliary carcinomas subgroup  and 9 in the gastrointestinal carcinoma subgroup . Both cancer types exhibited a significantly decreased survival rate in patients with hyper-expressed mucin 5ac. This may be because mucin 5ac can protect cancer cells from immune system attacks (the TRAIL-induced death pathways).27 However, researchers also noted that mucin 5ac has no effects on in vitro cell growth, cell survival, proliferation, or morphology,28 and studies on mucin 5ac function are relatively few. More function studies are needed to explore this finding. Second, in the Asian subgroup, 13 studies were included  and demonstrated that mucin 5ac may play a greater role in Asian populations. In the non-Asian subgroup, 4 studies were included , which may be a result of a genotype difference and/or environmental exposure or small samples in the non-Asian subgroup. Third, to clarify the prognostic value of mucin 5ac expression detection methods, we found statistical significance in the IHC  and IB  subgroups, but we should also note that as the 3 studies that did not use IHC were all performed in biliary tract cancer patients, it is hard to draw conclusions about the IB subgroup. In the ELISA subgroup, we did not find a significant association, which may be because only 1 study was enrolled and included only 33 patients. Therefore, the major methods (IHC and IB) to detect mucin 5ac expression might be efficient for predicting cancer patients’ prognoses.In the analysis of enrolled studies, we successfully drew some conclusions for clinical application. We observed that high mucin 5ac expression was significantly associated with a poor prognosis , which form a barrier that protects the epithelial cells.33 Tandem-repeats (TRs) are one of the hallmarks of mucins, which are rich in serine, threonine, and proline residues. Tandem-repeats are highly O-glycosylated and vital to mucin structure and function, and have been shown to be involved in specific ligand–receptor interactions.3 Mucins have also been shown to capture and hold biologically active molecules and antibodies, which, when released, might trigger inflammation, repair, or healing processes.34 Moreover, mucins are abnormally expressed in various cancers, and a previous report suggested that alterations in epithelial mucin core protein and glycosylation play an important role in cellular growth, differentiation, invasion, and immune surveillance for a variety of cancers,31 suggesting their potential role as promising biomarkers.36Mucins are heavily glycosylated proteins that are expressed by various epithelial cell types existing in relatively harsh environments (the air–water interface of the respiratory system, the acidic environment of the stomach, the complex environment of the intestinal tract, and secretory epithelial surfaces of specialized organs such as the liver and pancreas39 and has also been observed by us in one of our unpublished studies, with increasing occurrences of lymph node and distant metastasis and deeper invasion.40 Moreover, a 2013 study showed mucin 5ac could protect cancer cells from neutrophils’ attacking by suppressing TRAIL-mediated apoptosis .27 Therefore, mucin 5ac may serve as an important indicator in cancer prognosis.Mucin 5ac, a secreted gel-forming mucin that is secreted by goblet cells, has been shown to be expressed in higher levels in adenocarcinomas than squamous carcinomas. The overexpression of mucin 5ac in cancer has been documented by other researchers 41 so more studies focused on the aberrant glycosylation of mucin 5ac are also needed. Third, we extracted data from survival curves in some enrolled studies because the survival data were not presented directly, and these calculated HRs and 95% CIs might be less reliable than the directly given data. Fourth, the applied methods for detecting mucin 5ac expression and the cut-off values were different in the enrolled studies, which could cause heterogeneity among the studies.Although this study is the first meta-analysis of the association between MUC5AC expression and patient survival, some limitations of this meta-analysis should be acknowledged. First, our meta-analysis did not include all human tumor types. Although mucin 5ac is a promising biomarker, the correlation still requires further research. For further confirmed results, large-scale studies are needed. Second, most of the included studies used IHC to determine mucin 5ac levels, and because it is difficult to follow the same protocol in every study, a technique bias may exist. Moreover, aberrant glycosylation of mucins is observed in various cancers, and this may influence antibody recognition,This is the first meta-analysis evaluating the role of mucin 5ac as a cancer prognostic. Our study indicates that mucin 5ac may serve as a promising prognostic factor in cancer patients, especially in biliary carcinoma and gastrointestinal carcinoma patients, and is associated with lymphatic metastasis, which may play a more important role in Asian populations. More RCTs and functional studies are needed to explore the molecular mechanisms of mucin 5ac.