Both within-device and between-device reproducibility were lower than 9%, revealing satisfactory operational and fabrication reproducibility. Furthermore, the 3D-printed device was employed for the voltammetric determination of Colc in pharmaceutical tablets and in human urine with satisfactory results, justifying its suitability for low-cost routine analysis of Colc.In this work, a simple, fast, and sensitive voltammetric method for the trace determination of the alkaloid drug colchicine (Colc) using a 3D-printed device is described. The electrochemical method was based on the adsorptive accumulation of the drug at a carbon-black polylactic acid (CB/PLA) working electrode, followed by voltammetric determination of the accumulated species. The plastic sensor was printed in a single step by a low-cost dual extruder 3D-printer and featured three CB/PLA electrodes  and a holder, printed from a non-conductive PLA filament. The electrochemical parameters that affected the response of the device towards Colc determination, such as accumulation time and potential, solution pH, and other variables, were optimized. Under the selected conditions, the oxidation current of Colc was proportional to the concentration of Colc, and its quantification was conducted in the concentration range of 0.6–2.2 μmol L Colchicum autumnale L.). Its chemical structure is shown in Colchicine (Colc), N- acetamide, is a member of the proto-alkaloids group, prepared from the dried corns and seeds of the liliaceous plant meadow saffron  causes a reduction in pain and gout symptoms, whereas a high dose of Colc (4.8 mg over 6 h) can cause common side effects, such as gastrointestinal upset including nausea, diarrhea, vomiting, low blood cell count, and rhabdomyolysis, as well as bone marrow damage, anemia, and hair loss. Colc causes oxidative stress in animals, leading to cognitive impairment, and its therapeutic use has been linked to sporadic Alzheimer’s disease in humans  /K4[Fe(CN)6] as a redox probe. As shown in the cyclic voltammograms (ct), which was due to its relatively low conductivity. On the other hand, at the electrochemically activated 3D-printed WE (black trace), the Rct decreased significantly, suggesting that the charge transfer resistance of the electrode was lowered and the conductivity enhanced. Hence, the significantly reduced Rct can be attributed to the electro-etching effect induced by the electrochemical activation, which provides a faster electron transfer rate compared to the as-printed WE [−1 KCl solution containing 10 mmol L−1 K3[Fe(CN)6]/K4[Fe(CN)6]  was studied using cyclic voltammetry (CV) and differential pulse voltammetry (DPV) in 0.1 mol Lmmograms A and difmmograms B, when tmmograms , the resmmograms , the senmmograms A revealeinted WE . This faFe(CN)6] D, which The electrochemical method for the determination of Colc was based on the adsorptive accumulation of the drug on the surface of the electrochemically activated 3D-printed electrode, followed by the anodic voltammetric determination of the accumulated species. The adsorptive voltammetric response was evaluated with respect to the pH of the working solution, the voltammetric waveform, and the accumulation time and potential.−1 PB with pH 5.0, 7.0, and 9.0. As depicted in −1 PB with pH 7. 0, which was finally selected. The oxidation peak current shifted to more negative potentials by increasing the pH of the supporting electrolyte, and the inset of pa) and the pH with a slope of 0.053 (V/pH), which is close to the anticipated theoretical value of 0.059 for the electrochemical reaction of Colc, involving two protons and two electrons /K4[Fe(CN)6] as the electrochemical probe. The frequency ranged from 105 to 1 Hz with the application of 0.1 V of potential (DC) and 0.01 V of amplitude (AC). For the differential pulse voltammetric analysis of pharmaceutical tablets, 4 tablets of COLCHICINA/ACARPIA (1 mg Colc per tablet) were pulverized and dissolved in 100 mL of doubly distilled water. Next, 100 µL of the resulting solution was transferred into the electrochemical cell, which had previously been filled with 9.9 mL of 0.1 mol L−1 PB (pH 7.0). The urine sample was obtained from a healthy 22-year-old male volunteer with his informed written consent, and the measurements were conducted in accordance with GCP regulations. The urine sample was spiked with Colc to a final concentration of 10 µmol L−1 Colc, and then it was diluted 1:10 with 0.1 mol L−1 PB (pH 7.0). In human urine and pharmaceutical tablets, the determination of Colc was carried out by the method of standard additions to minimize matrix effects. For the interference study, possible substances that can be present in pharmaceutical tablets and human urine, such as glucose, sucrose, urea, uric acid, lactic acid, and K+, Na+, Ca2+, NH4 +, Cl−, PO43−, and SO42− were added at a concentration of 100 µmol L−1 in 0.1 mol L−1 PB (pH 7.0) solution that contained 1 µmol L−1 Colc. All potentials at the 3D-printed device are referred to with respect to the carbon-black-loaded PLA reference electrode.The connection of the 3D-printed device to the portable potentiostat was accomplished with crocodile clips. For the differential pulse voltammetric measurements, the solution was stirred at 1000 rpm and a potential of −1.2 V for 120 s was applied to the working electrode, followed by a differential pulse scan in a static solution, and the voltammogram was recorded. The differential pulse parameters were: modulation amplitude, 50 mV; increment, 10 mV; pulse width, 75 ms; and pulse repeat time, 50 ms. The cyclic voltammograms were obtained in 0.1 mol L−1) and reproducibility (%RSD < 9%) and allowed the determination of Colc in real samples with satisfactory selectivity and without the requirement of any separation or complex sample pretreatment. These features made the proposed method of utilizing the 3D-printed device practical for the simple and low-cost routine determination of Colc.In summary, a 3D-printed device consisting of three carbon-black-loaded PLA electrodes was successfully applied to the DPV quantification of the drug Colc. The electrochemical device exploited the advantages of fused deposition modeling in terms of fabrication speed and operation simplicity, while electrochemical activation via anodic and cathodic polarization significantly improved the electrochemical response of the device towards Colc. The voltammetric method offered excellent sensitivity (LOD of 0.11 μmol L


However, the combined effects of typhoons and upwelling have been less studied in the South China Sea. Based on satellite remote sensing data, we investigated potential contributions of temperature-characterizing upwelling and typhoon events to Chl-a changes in the Hainan northeast area. Results showed that the Chl-a concentration was 0.80 mg m-3 at the coastal upwelling index (CUI) of 1.7°C in the summer of 2020 when there were no typhoons crossing the area. The CUI (1.01°C) of typhoon-influenced period in 2019 was 0.21°C higher than that of typhoon-free period in 2019. And the Chl-a also increased from 0.70 mg m-3 to 0.99 mg m-3. In comparison, during the typhoon-free period, with the higher CUI, there was the higher concentration of Chl-a. In addition, the typhoon affected Chl-a concentration is significantly higher than that in the other two typhoon-free periods of 2019 and 2020. Though the typhoon has a limited effect on the upwelling intensity, the Chl-a concentration is much higher than when the upwelling acts alone. This is due to the combined effect of typhoon  and upwelling. The above results indicate that upwelling dominated the changes in Chl-a concentration in the Hainan northeast upwelling area during the typhoon-free period. In contrast, strong vertical mixing and runoff dominated the changes of Chl-a concentration during the typhoon-influenced period in the above area.Typhoons or upwelling are thought to promote higher phytoplankton chlorophyll-a (Chl- In summer, the stable stratification of the upper ocean leads to the limited nutrient supplement and lower biological productivity in the South China Sea. However, significant Chl-a increases are observed in coastal areas of Hainan, especially northeast of Hainan Island, which is mainly contributed by summer upwelling [Upwelling is the vertical upward movement of a water mass that carries cold and high nutrients water from the subsurface layer to the surface layer. And it plays an essential role in phytoplankton primary productivity, fishery production and even the ecosystem of the local area . The Soupwelling .a in nearshore is influenced by land-runoff, extreme weather, upwelling, monsoons, and currents [Phytoplankton Chl-currents –10. Upwecurrents  and booscurrents . Thus, ucurrents .a [a on the east side of Hainan Island change significantly [The coastal upwelling areas of Hainan Island usually has lower seawater temperature and higher phytoplankton Chl-a . Upwellia , 15. Fura . Typhoona . In addificantly .a [www.typhoon.org.cn). Thus, this study first compares the environmental elements characteristics of Hainan Northeast Upwelling (HNEU) in the summer of 2020 without typhoons to the typhoon-influenced climatology, using satellite remote sensing data. Then, the variabilities of sea surface temperature (SST), wind field (WS) and phytoplankton Chl-a concentration in the coastal upwelling zone of northeastern Hainan Island in summer in the presence and absence of typhoon years (months) are explored. Finally, the possible roles of temperature-characterizing upwelling and typhoon events in phytoplankton Chl-a variations are revealed in this area. This article would enhance understanding of co-effect of typhoons and upwelling in nearshore areas on phytoplankton, and provide the evidences for further exploring roles of interaction between typhoons and upwelling played in the ecological impacts of typhoons in the upwelling zone.It has been shown that the temperature drops of 2–3°C in the coastal upwelling area caused by tropical cyclone is significantly weaker than that in the non-upwelling area (the maximum temperature drops to 7°C in other areas) . When tha . The stua . Howevera data was carried out to evaluate the potential mechanism of typhoon or typhoon-free on phytoplankton Chl-a in the HNEU area in summer.The HNEU is located between 19.5–20.5°N and 110.5–111.5°E . This rea data from HERMES (https://hermes.acri.fr/) were utilized in this study to examine the spatial distribution characteristics of phytoplankton Chl-a in the HNEU area. The data are derived from Sea-Viewing Wide Field-of-View Sensor , Moderate Resolution Imaging Spectroradiometer , Medium Resolution Imaging Spectrometer , and Visible Infrared Imaging Radiometer Suite  instruments. The data uses the Garver-Siegel-Maritorena semi-analytical ocean color algorithm [a data have been widely used to study the distribution characteristics of phytoplankton Chl-a in the South China Sea [a product.The merged daily Chl-lgorithm . The Chlhina Sea . Considehttps://www.ncei.noaa.gov/) with a spatial resolution of 0.25° and a temporal resolution of 1 day. The time span from September 1981 to the present. The sea surface wind speed (WS) in our research region was mapped using the Cross-Calibrated Multi-Platform (CCMP) product, which is created by Remote Sensing Systems (https://www.remss.com/) and is accessible every 6 hours with a spatial resolution of (0.25° × 0.25°).Sea surface temperature data are provided by the Optimum Interpolation Sea Surface Temperature (OISST) product from the National Centers for Environmental Information (https://www.typhoon.org.cn/) [The typhoon data is an online typhoon best track dataset provided by China Typhoon Network (org.cn/) , which cThe Coastal Upwelling Index (CUI) is commonly used to define the intensity of upwelling . Ru ZhanSSToffshore is the sea temperature in the non-upwelling area at the same latitude as the upwelling center, and SSTcoast is the temperature of the upwelling center. The difference between the two is taken to characterize the intensity of the upwelling. In this study, the area of 400 nautical miles from the upwelling area at the same latitude is defined as the non-upwelling area.Where p < 0.05 to indicate statistical significance of the correlation coefficient [Pearson correlation is a statistical method used to evaluate the linear relationship between two variables . In thisfficient .-1–6 m s-1  was slightly lower than that in July 2019 (~ 1.11 mg m-3). In a concentration in most areas on the left side of the trapezoid in 2019 is about 0.5 mg m-3 higher than that in 2020. While the Chl-a concentration in the upper left part of the trapezoid in 2020 is significantly higher than that in 2019. It coincides with the position of the new upwelling center after moving northward  is also significantly higher than that in 2019 (0.70 mg m-3). It is worth noting that the results show an interesting phenomenon: the CUI in 2020 was higher than that during typhoon crossing in 2019, but the Chl-a concentration shows the opposite trend.Further reduce the time scale to the typhoon-influenced period. We selected the CUI and Chl-hoon MUN  impact pa during the typhoon-influenced period in 2019 (R2 = 0.7381) (a concentration was still low at this time (below the trend line in a increased at first and then decreased. The CUI is moderately positively correlated with Chl-a during the typhoon-free period in 2019 (R2 = 0.1208). The CUI has a strong positive correlation with Chl-a during the typhoon-free period in 2020 (R2 = 0.2635). In these two typhoon-free periods, the Chl-a concentration all increased with the increase of CUI. From the above correlation results, it can be seen that there is a direct relationship between Chl-a and upwelling, which will be changed by typhoon crossing.The CUI is highly negatively correlated with Chl- 0.7381) . Typhoona and CUI suggested the upwelling may control the distribution of phytoplankton Chl-a in the HNEU under the typhoon-free influence in July of both 2019 and 2020. In the typhoon-free period, Chl-a concentration become higher with increasing upwelling strength. The strong correlation  or without typhoon (i.e. 2020) were observed in HNEU area during summer. Though the CUI of HNEU area is significantly higher than the typhoon-influenced period for many years, the phytoplankton Chl-a concentration is lower, suggesting typhoon would exert an important factor for the above phenomena. Here, a schematic diagram of the ecological processes acting on the HNEU area with and without typhoon is presented (a concentration in HNEU area during the typhoon-free period (i.e. higher Chl-a with high CUI). Normally, Ekman pumping caused by wind stress curl plays a more important role in upwelling than Ekman transport by wind stresses parallel to the coastline. Response of phytoplankton Chl-a concentration to typhoons are more evident than that upwelling in HNEU area. The superposition effect of stronger sea water mixing by strong wind stress during typhoon and enhanced upwelling increases further the vertical nutrient transport Click here for additional data file.


The present study sought to examine the factor structure, reliability, validity and gender invariance of the Arabic Body Appreciation Scale (BAS-2), and its two short forms (the 3-item and 2-item BAS-2SF) among community Lebanese Arabic-speaking adults.We carried-out an online cross-sectional survey. The 10-item BAS-2, the functionality appreciation scale and the Eating Attitudes Test-26 were completed by 826 Lebanese community adults .The Arabic 10-item, 3-item and 2-item BAS-2 converged on a one-factor solution, and demonstrated high internal consistency . All indices suggested that configural, metric, and scalar invariance was supported across gender. The 3-item and 2-item BAS-2SF were highly correlated with the original BAS-2 (r > 0.9). Higher 10-item, 3-item and 2-item BAS-2 scores correlated significantly and strongly with more positive eating attitudes and greater functionality appreciation, and higher 10-item BAS-2 scores correlated positively and weakly with BMI, supporting convergent validity.The present work contributes to the literature by providing a psychometrically sound Arabic-translation of the BAS-2 and short-forms, benefitting both researchers and clinicians.The online version contains supplementary material available at 10.1186/s40337-023-00885-x. Research on body image traditionally has focused on describing and predicting negative body image such as body dissatisfaction, body shame, and body preoccupation, with less focus on identifying, predicting, and promoting adaptive body attitude. Recently, however, the study of positive body image has gained considerable motion, and body appreciation has been the central organizing variable within these investigations. Body appreciation has been defined as accepting, holding favourable opinions toward, and respecting the body, while also rejecting media-promoted appearance ideals as the only form of human beauty. The Body Appreciation Scale has been utilized by researchers to understand features, correlates, and potential outcomes of positive body image. This scale and its short forms are not yet validated in Arabic. To that end, we aimed to translate and validate the Arabic version of the Body Appreciation Scale (BAS-2), and its two short forms (the 3-item and 2-item BAS-2SF) in the present study, which would in turn facilitate improved research and clinical practices related to body appreciation in Arabic-speaking nations. The present findings provide support for the psychometric properties of the Body Appreciation scales in Arabic-speaking adults in Lebanon.The online version contains supplementary material available at 10.1186/s40337-023-00885-x. Body image can be defined as individual’s perceptions, feelings, and thoughts about their own physical appearance , and encSince its original validation in 2015, the BAS-2 has shown its strong psychometric quality within various population-based samples of varying characteristics, including adolescents –14, studPositive body image concepts, including behavioural expressions toward the body, have been suggested to depend on cultural norms and values, and therefore to express differently across societies, cultures and religions –38. For In this paper, we propose to test the psychometric properties of an Arabic translation of the BAS-2 and its short forms (the 2-item BAS-2SF and the 3-item BAS-2SF) in a sample of community adults within an Arabic-speaking context, Lebanon. Even though an Arabic version of the scale exists, its validation was limited in certain ways. First, authors relied on a college student sample, which implies that findings may not be generalizable to the wider community. Second, only women have been involved in this validation study, which did not allow for comparisons across genders and for gender invariance to be ascertained. This represents a major gap, given that invariance across women and men of BAS-2 scores yielded mixed findings in previous validations across countries  the Arabic full-length and short forms of the BAS-2 would have a good internal consistency; (2) our analyses would endorse a good fit of a unidimensional factor structure and show measurement invariance across gender of all three forms of the scale; (3) an examination of the correlations between BAS-2 and BAS-2SF scores, disordered eating attitudes, functionality appreciation, and BMI would provide evidence of convergent validity; and (4) both Arabic BAS-2SF versions’ scores would strongly correlate with the BAS-2 total scores, suggesting that the two short forms evaluate the same body appreciation construct.SD = 8.44) and 57.9% females. Other characteristics are displayed in Table A total of 826 Lebanese citizens and residents enrolled in this study with a mean age of 25.42 years . Higher scores on this scale reflect greater body appreciation. Following international guidelines [This 10-item instrument assesses acceptance of one’s body, respect and care for one’s body, and protection of one’s body from unrealistic beauty standards (BAS-2 ). Items idelines , the scaidelines . In the strongly disagree to strongly agree) [The FAS was used to assess how respondents appreciate, respect, and honour their own bodies for what it is capable of doing. The scale is a unidimensional scale, composed of seven items  rated on a 5-point scale (y agree) . Higher y agree) , which dnever) to 6 . The scale is unidimensional, with higher total scores reflecting greater disordered eating attitudes. In the present study, the Cronbach’s alpha was 0.90.Participants were asked to complete the Eating Attitudes Test-7 , whichParticipants were asked to provide their demographic details consisting of age, sex, and education level.2.Self-reported data about height and weight were used to compute BMI in kg/mEthics approval for this study was obtained from the Psychiatric Hospital of the Cross ethics committee . All data were collected via a Google Form link, between December 2021 and April 2022. The project was advertised on social media and included an estimated duration. Inclusion criteria for participation included being of a resident and citizen of Lebanon of adult age. Internet protocol (IP) addresses were examined to ensure that no participant took the survey more than once. After providing digital informed consent, participants were asked to complete the instruments described above, which were presented in a pre-randomised order to control for order effects. The survey was anonymous and participants completed the survey voluntarily and without remuneration.2/df), the Steiger-Lind root mean square error of approximation (RMSEA), standardized root mean square residual (SRMR), the Tucker-Lewis Index (TLI) and the comparative fit index (CFI). Values ≤ 5 for χ2/df, ≤ 0.05 for SRMR, ≤ 0.08 for RMSEA, and 0.90 for CFI and TLI indicate good fit of the model to the data [There were no missing responses in the dataset. To examine the factor structure of the BAS-2, we conducted a Confirmatory Factor Analysis (CFA) on the total sample using the SPSS AMOS v.29 software. A previous study suggested that the minimum sample size to conduct a confirmatory factor analysis ranges from 3 to 20 times the number of the scale’s variables . Therefothe data . Additiothe data . MultivaTo explore the factor structure of BAS-2-SF (2 and 3 items), we computed a principal-axis EFA with the first split-half subsample using the FACTOR software . We verit-test only if scalar or partial scalar invariance were established.To examine gender invariance of BAS-2 scores, we conducted multi-group CFA . MeasureComposite reliability in both subsamples was assessed using McDonald’s ω and Cronbach’s α, with values greater than 0.70 reflecting adequate composite reliability. Normality was verified since the skewness (= − 0.511) and kurtosis (= − 0.354) values for the total score varied between − 1 and + 1 . To asse2/df = 177.32/35 = 5.07, RMSEA = 0.070 , SRMR = 0.018, CFI = 0.982, TLI = 0.976. The standardised estimates of factor loadings were all adequate  compared to men , t(824) = -5.337, p < 0.001, d = 0.372.As reported in Table The 3 items  and the 2 items (items 6 and 9) converged on a one-factor solution, with high loadings in the total sample and in men and women separately. Plus, the reliability values were very good (> 0.8) Table .Table 4ITo assess the validity of the three BAS-2 scores, we examined bivariate correlations with all other measures included in the present study using the total sample. As can be seen in Table Despite their peculiarities, Arabic-speaking people have long been neglected in the body image research literature due to a lack of valid and adapted screening tools . While tUsing EFA and CFA, our findings demonstrated that the unidimensional factor structure of the three Arabic forms had a good fit to the data in both genders. This is in agreement with the original English validation of the 10-item BAS-2, where all 10 items loaded onto one factor in both men and women respondents . This isAll three Arabic forms of the BAS-2 also evidenced internal consistency and invariance by gender at the metric, configural, and scalar levels. This is in line with findings from the original BAS-2 and BAS-SF validation studies , 34, as Our findings have to be carefully interpreted bearing in mind certain limitations. The test–retest reliability of the BAS-2 and BAS-2SF was not examined in the context of the present study, precluding any conclusions regarding scores stability across time. In addition, while we provide support for the validity of these Arabic versions in a community sample of Lebanese adults, future research need to assert their psychometric robustness in clinical samples, and samples from Arab countries others than Lebanon. Cross-cultural research need also to ascertain the invariance of these measures across Arab countries and communities. Another limitation is the self-report nature of all measures used in our study that might compromise the accuracy of responses due to social desirability bias.While one's relationship with body image is constantly evolving and changing in the Arab world as a result of the globalization process, research in this field has not followed at the same pace. One major and unique contribution of the present study was the examination of gender invariance and comparison of the body appreciation construct across genders in an Arab context. Another contribution was making shorter forms of the BAS-2 available for Arabic-speaking populations. Due to their brevity and good psychometric properties, the Arabic 3-item and 2-item BAS-2SF are advantageous for assessing body appreciation in Arab clinical and research settings, especially when long questionnaires at multiple time points are needed . We thus hope that providing these valid, simple, easy-to-use and low-cost tools will be of substantial benefit to clinicians, and will stimulate more research in this area in the Arab context. Such research may inform strategies aiming at promoting the appreciation and respect of one’s body in Arabic-speaking communities worldwide.Additional file 1. Body Appreciation Scale-2.


The limits of detection and quantification were within the ranges of 0.6–40 µg/L and 2.0–135 µg/L, respectively, with the exception of spermine (SPM). The recoveries were demonstrated within the range of 82.6–103.0%, with relative standard deviations (RSDs) of less than 4.2%. This simple method featuring excellent sensitivity and selectivity was suitable for the quantification of BAs in wines. The occurrence of BAs in 236 wine samples that are commercially available in China was determined. The BA levels in wines of different geographical origins varied significantly. The acute dietary exposure assessment of BAs was carried out by calculating the estimated short-term intake (ESTI) and comparing the acute reference dose (ARfD) specified by the European Food Safety Authority (EFSA). Results showed that the exposure to histamine (HIS) and tyramine (TYR) via the consumption of wines was much lower than the recommended ARfD level for healthy individuals. However, exposure could lead to symptoms in susceptible individuals. These results provided basic data regarding the occurrence and risk of BAs in wines for wine production, health guidance and consumer safety.A method for the determination of nine biogenic amines (BAs) in wine was established using ion chromatography–tandem mass spectrometry (IC-MS/MS) without derivatization. BAs were separated by a cation exchange column  with a gradient aqueous formic acid elution. Good linearity was obtained for nine BAs with coefficients of determination (R Biogenic amines (BAs) are basic, non-volatile, nitrogenous organic compounds of low molecular weight . AccordiHigh levels of BAs may lead to symptoms such as headache, hypertension, diarrhea, nausea, red rash and inflammation. HIS and TYR are considered the most harmful BAs, and they can act synergistically with PUT and CAD to aggravate poisoning symptoms, especially for susceptible individuals . The conThe presence of BAs in wine is common and inevitable . There aIt is widely recognized that BAs that are present in foods or beverages may trigger toxic symptoms. However, there is no shared legislation to limit the content of BAs in wine . The recThere have been a large number of studies carried out on BAs in wine from different countries, and these mostly aim to monitor the safety and quality of wine. However, there are only a few studies on BAs in wine with the purpose of determining the geographical origin or grape variety. Mitar et al. determined the contents of BAs in Croatian wine from two different wine-producing regions and found that the levels of BAs were associated with the geographical origin . Ladenteo-phthalaldehyde [Numerous analytical methods have been reported for the determination of BAs, such as HPLC, CE, GC and IC. HPLC coupled to UV ,23 or flaldehyde , due to aldehyde . In contaldehyde . In thisaldehyde  using inaldehyde  and BAs aldehyde . AccordiIn this work, a simple and rapid method for the determination of nine BAs in wine was developed. The separation of analytes was achieved using cation exchange chromatography, and the detection was performed by tandem mass spectrometry (MS/MS). This method not only simplified the sample’s pretreatment procedure without derivatization but also achieved good separation as well as high sensitivity and selectivity using MS/MS detection. Considering the great interest in the risk assessment of BAs in wine, the occurrence of BAs in 236 commercially available wine samples of different geographical origins in the Chinese market was investigated. This study may enrich the knowledge of the analytical methods of BAs and will be useful for the supervision of wine quality and safety, and it can establish the maximum allowable limit of BAs for wine in China.4H12N2⋅2HCl, >98%), cadaverine dihydrochloride , histamine dihydrochloride , tyramine hydrochloride , spermine tetrahydrochloride , spermidine trihydrochloride , tryptamine hydrochloride , octopamine hydrochloride  and 2-phenylethylamine  were purchased from Shanghai Yuanye Bio-Chem Technology Co., Ltd., . Formic acid was of HPLC grade and purchased from Tedia Company . Ultrapure water (18.2 MΩ/cm) was prepared by a Millipore-Milli-Q system . The other chemicals used in this study were of analytical grade.Putrescine dihydrochloride (Cn = 135), followed by France (n = 22), Australia (n = 19), Italy (n = 18) and other countries . The origin of the white wine samples comprised Italy (n = 11), followed by the Czech Republic (n = 6) and China (n = 6). The majority of red wine was produced with grape cultivars comprising Cabernet Sauvignon, Marselan, Cabernet Gernischet, Merlot, Syrah and Cabernet Franc grape varieties. Regarding the white wine, most were produced with the Chardonnay grape variety.A total of 236 commercial wine samples produced in different countries were collected and analyzed in this study. Of these wine samples, 213 were red wine samples and 23 were white wine samples according to the description on the label of the bottles. The origin of the red wine samples comprised China , and detection was performed with a tandem QTRAP5500 mass spectrometer . The used column was a cation exchange guard column  with a column temperature of 35 °C. The eluent consisted of 1.0 M aqueous formic acid (A) and ultrapure water (B). The linear gradient elution was as follows: 0.0 to 15.0 min, a linear gradient from 3% to 25% A; 15.0 to 16.0 min, a linear gradient from 25% to 100% A; 16.0 to 25.0 min, 100% A; 25.0 to 26.0 min, a linear gradient from 100% to 3% A; 26.0 to 30.0 min, 3% A. Analysis time was 25 min, and an additional 5 min was required for system equilibrium. The flow rate was 1.0 mL/min, and a flow divider valve was used before MS detection. The injection volume was 10 μL. MS analysis was carried out in the multiple-reaction monitoring (MRM) mode. Electrospray ionization was carried out in the positive ion mode. The MS parameters were as follows: ion spray voltage of 5500 V; gasification temperature at 500 °C; curtain gas at 40 psi; ion source gas at 50 psi and auxiliary gas at 60 psi. The conditions for MRM  were optimized for each BA. The MRM transitions are listed in 2) were calculated from the calibration curves. The method’s sensitivity was evaluated using the limits of detection  and limits of quantification . Repeatability was tested by injecting the standard working solution six consecutive times. Reproducibility was evaluated by analyzing a spiked wine sample six consecutive times. The accuracy was determined via the triplicate analysis of spiked wine samples at different spiking levels.The proposed method was validated by evaluating the validation parameters, including linearity, sensitivity, precision and accuracy. Linearity was assessed using calibration curves obtained after the injection of the mixed standard solutions of BAs with a wide concentration range. The coefficients of determination  of HIS and TYR was calculated and compared with the acute reference dose (ARfD) specified by EFSA . To evalThe exposure of BAs was calculated as follows:ESTI = C × QESTI = Estimated short-term intake .C = The 50th and 95th percentile contents of BAs in wines (mg/L).Q = Individual wine consumption at the 95th percentile .Microsoft Excel software was used to calculate the P5, P95, mean, median, max, standard deviation and RSD values. Principal component analysis (PCA) was performed using Origin 2018 software .2SO4 and methanesulfonic acid, some BAs were hardly eluted even with high concentrations of formic acid in the eluent. In order to obtain rapid analysis times, only the short CG17 guard column was used for the separation. The separation of nine BAs was achieved using a gradient elution of formic acid solution. The direct injection of the individual standard solution (1 mg/L) of each BA was performed to optimize mass conditions. Mass scans were carried out in the positive ion mode. After confirming the Q1 mass, two characteristic ion pairs were selected as qualitative and quantitative ions for each BA using a Q3 mass scan. Ionization conditions and collision energy were optimized for the best sensitivity  ranging from 0.9972 to 0.9998. The LODs and LOQs for eight BAs (except SPM) were within the ranges of 0.6–40 μg/L and 2.0–135 μg/L, respectively. Relatively high LODs (500 μg/L) and LOQs (1700 μg/L) were obtained for SPM due to the high concentration of the eluent. The repeatability of the developed method provided RSD values ranging from 0.9% to 2.4%, whereas the reproducibility of the method showed RSD values ranging from 0.6% to 3.4%. Good reproducibility results were probably due to the simple sample pretreatment with only one dilution step. The recovery percentage ranged from 82.6% to 101.9%, from 83.6% to 99.5% and from 84.9% to 103.0% in a selected wine sample using three different spiked levels, with RSD values lower than 4.2%  interferes with the determination of BAs. In addition, some BAs with low pKa values have difficulty forming cations and cannot be detected by using a conductivity detector. Lately, there has been growing interest in the use of GC, HPLC and IC coupled to electrospray ionization (ESI) and mass spectrometry (MS). ESI ionizes the analytes directly prior to MS detection without derivatization treatments. Furthermore, the use of MS provides a significant improvement in the method’s sensitivity, selectivity and reliability of quantification. In this study, a reliable IC-ESI-MS/MS method was developed for the accurate determination of BAs present in wine. Compared with HPLC methods, the proposed method allowed the achievement of simple and eco-friendly procedures without using derivatization steps. Compared with IC-conductivity or UV detection, the analytical figures of merit obtained with the proposed method highlighted its competitiveness with respect to the number of determined BAs, sensitivity and selectivity. In this study, it can be observed in the validation results that the method showed a good linear range, repeatability, reproducibility and accuracy, which indicated that the proposed IC-MS/MS method was sufficiently suitable for the determination of BAs in wines.The grape variety, winemaking process, specific strains and the composition of free amino acids are the main factors affecting the contents of BAs in wine and cause substantial differences in red and white wine . Both maEsposito et al. evaluated the dietary exposure to BAs via the consumption of wine commercialized in Italy, and results showed that no exposure values exceeded the levels of ARfD  . Han et In this study, a simple, rapid and sensitive method for the determination of nine BAs in wines was established by IC-MS/MS without derivatization. The developed method showed excellent analytical performance in terms of linearity, sensitivity, precision and accuracy, indicating its suitability for the fast, accurate and environmentally friendly determination of BAs in wine. The occurrence of BAs in 236 wine samples available in the Chinese consumer market was investigated. Significant variations were found in the individual and total contents of BAs present in the wine of different geographical origins. The highest content of the individual BA was found to be that of PUT, followed by HIS, TYR and CAD in wines, proving that the introduction of BAI can be used to evaluate the quality of wine. The red wine samples investigated in this study showed higher BA levels than those found in the white wines. According to the risk assessment results of HIS and TYR, the exposure values were much lower than the levels of ARfD specified by EFSA. The wine available in the Chinese consumer market is considered safe for healthy individuals but may present risks in susceptible individuals with HIS intolerance or patients taking classical MAOI drugs. This study not only provided an efficient IC-MS/MS method for BA determination but also enriched information regarding the occurrence and risk of BAs in wine available in the Chinese market. All these are of great significance for wine production, health guidance and consumer safety.


Mage = 3.41) through first grade (Mage = 7.92). Results demonstrate that parent invitations for children to explain causal phenomena had strong concurrent relations to scientific literacy but showed little relation to subsequent literacy. In contrast, the broader home science environment at preschool entry, particularly in the form of exposure to science-related activities, predicted scientific literacy over the next 4 years. The directionality and specificity of these relations were clarified through the inclusion of measures of cognitive and broader home experiences as controls in regression analyses. Overall, our investigation revealed that exposure to science-related input provided by parents has particularly powerful potential for shaping scientific literacy when children are very young. Implications for parent-focused interventions that promote science literacy are discussed.Fostering scientific literacy has become an increasingly salient goal as evidence accumulates regarding the early emergence of foundational skills and knowledge in this domain, as well as their relation to long-term success and engagement. Despite the potential that the home context has for nurturing early scientific literacy, research specifying its role has been limited. In this longitudinal study, we examined associations between children’s early science-related experiences at home and their subsequent scientific literacy. Following on our previous work, we specifically considered parent causal-explanatory talk, as well as the degree to which parents facilitate access to science-related materials and experiences. A group of 153 children from diverse backgrounds were evaluated across 5 annual waves of data collection from preschool entry ( Although the majority of this work has centered around school-aged children or typically children 5 years and up in the USA e.g., , researcScientific literacy refers to our understanding of core disciplinary ideas  and practices , as well as cross cutting concepts  as reflected in the current Next Generation Science Standards . It is nAlthough the development of evidence-based preschool science curricula e.g.,  has beeninvitations for their 3-year-olds to generate causal explanations during free play correlate with the children’s concurrent scientific literacy.First, we consider the degree to which parents evoke causal explanations in conversations with their children. Causal explanations are descriptions of how or why factors in a system influence each other. When parents scaffold children’s consideration of causal explanations, it may support scientific literacy by both contributing directly to conceptual knowledge and by offering opportunities for practicing scientific inquiry processes like hypothesis generation and revision. Several studies have broadly observed the frequency and quality of conversations between parents and their children in a variety of informal science settings . For exaSecond, we consider the broader home science environment, including access to science-related materials and experiences provided by parents . EmpiricThis work aims to further clarify links between aspects of early experience and emergent scientific literacy by examining four years of longitudinal data beyond the initial wave reported in We then ask whether broader indicators of the home science environment are potentially foundational to early scientific literacy. In addition to the composite measure of home science used in Throughout, we capitalize on the longitudinal nature of the data to achieve greater precision in our conclusions. Specifically, we clarify the directionality of effects by controlling for initial scientific literacy. We also clarify the specificity of observed associations by controlling for general cognitive skills and broader (non-science) cognitive stimulation in the home, and by considering contrastive predictions to math and reading skills.2.2.1.a priori power analysis using G*Power software . Child participants were described by their caregivers as proficient in English and absent of any diagnosed developmental delay or disorder. At the first session, eight additional children were excluded based on their inability to follow instructions due to inadequate English knowledge or behavioral noncompliance. The sample was demographically diverse across race, ethnicity, and maternal education  remained in the study, at the third wave 112  remained, at the fourth wave 88  remained, and at the final wave 87 . Throughout these years of assessment, attrition was primarily due to families moving out of town or our inability to re-establish contact. Data collection at the fourth wave was also substantially disrupted by the COVID-19 pandemic, resulting in a spike in attrition and an eventual shift to virtual data collection  and lasted between 30 and 60 min. All Wave 1, 2, and 3 sessions were conducted in a laboratory setting except for the very first session of Wave 1 which was conducted at a local science museum. Although testing began in the laboratory for Wave 4, the global pandemic necessitated that we shift to virtual format. Wave 5 was also conducted virtually. After each session, caregivers received financial compensation and, if the session was run in person, the child was also given a book.2.3.Our investigation included measures of parent causal talk and the broader home science environment, as well as children’s scientific literacy and general cognitive ability. See 2.3.1.2.3.1.1.Parent–child dyads played freely with toys affording causal explanations at Waves 1 (sink-float task in the lab and a launcher museum exhibit that involved building and testing airplanes), 2 (Hasbro’s Mouse Trap™ game) and 3 . Based on pilot observations and timing constraints, participants were allotted 10 min to play with each set of toys. They were given no specific instructions about how to interact with the toys and were only told to let the experimenter know if they wanted to stop early. The 10 min of play were coded offline and broken into 60 s windows. For each window, coders indicated if the parent (1) produced a causal explanation, (2) invited the child to explain a causal phenomenon, and (3) provided any other causally relevant utterance. Utterances coded as causal explanations often included “because” or “if, then” statements, such as, “If I put the ball here, then this will make it fall.” Likewise, utterances coded as causal invitations often contained “why” or “how,” such as, “Why do you think that one sank?.” See For each of these causal constructs, the proportion of 60 s windows  in which parents produced at least one utterance of each target construct was calculated. Utterances were coded as causally oriented even if the information provided in that utterance was not technically correct . The proportion of windows in which parents specifically produced an explanation or an invitation served as the key dependent variables. Note that because two play sessions were implemented at Wave 1 (in the lab and museum), final proportions were attained by averaging across contexts. An additional research assistant independently coded 20 percent of the co-play sessions. Although reliability was generally good , discrepancies were jointly reviewed to arrive at full consensus.2.3.1.2.A parent survey was administered at each wave of data collection asking about the number of science-themed books, toys, and apps/computer games available in the home. No specific examples were provided for these home resources and it was left to the caregiver to determine what counted as “science.” These responses were each coded into bins  and then summed across types to calculate a ‘materials’ score ranging from 0 to 12. Parents were also asked about the frequency  with which they participate in science activities  with their child, as well as how often they visit science fairs or museums with their children . These were then summed into an ‘activities’ score ranging from 0 to 12. The material and activities scores were then summed into the total home science environment score ranging from 0 to 24. See 2.3.1.3.The StimQ-Preschool  scale me2.3.2.Different developmentally appropriate measures of scientific literacy were utilized at each measurement time point. More information on the scientific literacy measures is available on OSF.2.3.2.1.Lens on Science  is an ad2.3.2.2.The SLA  is desig2.3.2.3.TerraNova Science Subtest  is a sta2.3.2.4.Administered at the last wave, the SKI  consists2.3.3.2.3.3.1.The ECB  is a hig2.3.3.2.As with TerraNova Science Subtest, the Math Subtest  is a nor2.3.3.3.The TORTT  is an ad3.χ2 (3441) = 3450.59, p = 0.404. To address missing data in a maximally unbiased manner, we conducted 100 iterations of multiple imputation including our home science environment and scientific literacy measures. Demographic and cognitive variables that correlated highly (r > 0.40) with our key measures (and had no more than 25% missingness themselves) were also included as auxiliary variables in the imputation . This decision was based on the face validity of conceptual equivalence across our scientific literacy measures and the significant correlations between them at each measurement wave see . Next, w3.1.r = 0.24, p = 0.006). Although parent invitations to explain at Waves 2 and 3 failed to correlate with either concurrent or subsequent scientific literacy, parent invitations to explain at Wave 1 did also correlate with scientific literacy one year later . In contrast, parent-produced causal explanations failed to correlate with scientific literacy in any analysis , p = 0.265.Given that (1) parent invitations-to-explain correlated with scientific literacy at Wave 1 and (2) scientific literacy was stable across Wave 1 and 2, it was important to clarify whether the observed association between parent invitations-to-explain at Wave 1 and scientific literacy at Wave 2 was truly predictive (rather than merely a spurious byproduct of these other two associations). We therefore conducted a multiple regression analysis, controlling for baseline (Wave 1) scientific literacy scores. Consistent with an early association carried forward by stability in measurement of scientific literacy, parent invitations were no longer predictive of scientific literacy scores at Wave 2 in this analysis, 3.2.r = 0.18, p = 0.028) and Wave 2 , and Wave 3  scientific literacy scores. The home science environment at Wave 2 was significantly positively related to scientific literacy at all waves, with the strongest associations observed at Wave 4  and Wave 5 . Although later home science environment scores were less strongly correlated with scientific literacy scores in general, science environment scores at Waves 3, 4, and 5 were significantly associated with scientific literacy at Wave 5 , p < 0.01 and Wave 3, B = 0.06 (SE = 0.10), p < 0.01, over and above baseline scientific literacy. To explore the possibility that the observed relation between home science environment and subsequent scientific literacy was due to a common reliance on broad cognitive skills, we ran a follow-up regression analysis including NIH-ECB as an additional predictor. Indeed, when controlling for baseline cognitive skills, the Wave 1 home science environment no longer accounted for a significant amount of variance in Wave 2 scientific literacy scores, B = 0.04 (SE = 0.02), p = 0.076, although a trend was still evident. The home science environment did, however, predict Wave 3 scientific literacy scores, B = 0.05 (SE = 0.02), p < 0.05 over and above baseline cognitive skills . To this end, we first ran a multivariate regression analysis with home science environment at Wave 1 predicting scientific literacy at Waves 1, 2, and 3, while controlling for baseline (Wave 1) scientific literacy see , Model 1ills see , Model 2B = 0.06 (SE = 0.03), p < 0.05, and trending toward significance for Wave 5 but not at Wave 3. When Wave 2 cognitive skills are added into the regression, the Wave 2 home science environment no longer accounts for significant variance in any subsequent scientific literacy score, although trends remain near significance for Waves 4 and 5.We then turned our attention to the relation between Wave 2 home science environment and subsequent scientific literacy scores. Again, we added baseline scientific literacy score as a predictor in the regression analysis see , Model 1ps > 0.10).Although the analyses reported thus far are consistent with at least some truly predictive relations between the early home science environment and subsequent scientific literacy, it is still possible that the reciprocal relation exists whereby a child’s level of scientific literacy might shape their subsequent home literacy environment. Some significant bivariate correlations seen in activities at Wave 1 and scientific literacy measured concurrently at all subsequent waves. Science materials, in contrast, only correlated relatively weakly with scientific literacy at the second wave of measurement. These associations reversed somewhat when the home science environment at Wave 2 was instead considered. Here, science activities only correlated significantly with scientific literacy at Wave 4 and 5 while science materials correlated significantly with all waves of scientific literacy, although only at a trend level for Wave 3. Correlations between later measures of home science activities and materials with scientific literacy were only sporadically observed.In further exploratory analyses, we examined the materials and activities subcomponents of the home science environment score separately. As can be seen in Importantly, when submitted to regression analyses including baseline scientific literacy see , Model 1ps > 0.10). None of the few reciprocal correlations between early scientific literacy and later home science activities or materials observed in ps > 0.10).In contrast, no predictive relations between access to science materials and subsequent scientific literacy hold when baseline scientific literacy is included as a control , it was not to the degree to cause a collinearity threat. The StimQ-P did not significantly predict scientific literacy at Wave 2, B = 0.02 (SE = 0.02), p = 0.409, nor Wave 3, B = −0.01 (SE = 0.02), p = 0.683. Home science environment  at Wave 1, on the other hand, continued to predict scientific literacy at Wave 3, B = 0.06 (SE = 0.03), p < 0.05, over and above StimQ-P (see B = 0.06 (SE = 0.03), p < 0.05. In addition, excluding the home science environment factor entirely  failed to yield any significant predictions to scientific literacy at any time point.Given that there are many ways that children’s home environment might contribute to the development of their scientific literacy, it was important to distinguish our measure of the home science environment from the broader richness of the home environment. We therefore added the StimQ-P, the more general measure of the home learning environment, as another predictor into our regression models. While scores on the StimQ-P positively correlated with our measure of the home science environment at Wave 1  taken at the last measurement time point. No associations between these outcomes and any aspect of early input  were detected see . We ther4.The goal of this longitudinal project was to broaden understanding of the relations between early science-related input and children’s emergent scientific literacy by building upon data first collected at 3 years of age and reported in With respect to parent causal-explanatory talk, we found that parent invitations to explain causal phenomena were related to children’s scientific literacy concurrently and one year later. However, because this singular longitudinal association failed to hold when controlling for initial scientific literacy, no clearly predictive relations were evident. It is entirely possible that, if a predictive relation between parent invitations to explain and scientific literacy exists at all, it is confined to only the very earliest developmental window. Any correlations to later scientific literacy might well be due to subsequent stability in outcome measurements. Notably, no associations between the degree to which parents provided explanations themselves and children’s concurrent or subsequent scientific literacy were observed at all. Although surprising given the theoretically plausible usefulness of these explanations for building scientific knowledge, this finding is consistent with results reported for the first wave of data collection in Together, these results suggest that parent causal-exploratory talk plays little role in shaping emergent scientific literacy. However, related research has found parent explanatory talk to be positively related to children’s concurrent science-related exploration and learning . StudiesOur failure to observe robust associations between aspects of parent causal talk and children’s scientific literacy might be due to limitations of our measurement approach. First, our observations were quite brief and constrained to laboratory settings (with the exception of the one museum observation at 3 years). While the play materials available in these contexts were intended to evoke natural conversations about causality, other or more varied options might have been more successful. More extended recordings in the home would have perhaps been most ideal for capturing natural variation. Second, our coding of the existing data did not capture potentially important nuances in the input. For example, we did not consider the quality of explanations produced by parents or children. Studies find explanations produced by parent and children are not always accurate or exhaustive , and it In contrast to the relatively weak effects observed in consideration of parent causal-explanatory talk, our analyses of the broader home science environment were much more promising. Specifically, we found the home science environment at Wave 1 to be related to contemporaneous scientific literacy, as well as to scores at subsequent Waves 2 and 3. These longitudinal relations held even when controlling for initial scientific literacy scores and could not be fully accounted for by general cognitive abilities. Similarly, the home science environment measured at Wave 2 predicted concurrent scientific literacy, as well as scores at all subsequent measurement waves. These relations variably held when baseline scientific literacy and cognitive skills were included as controls. Importantly, reciprocal relations between early scientific literacy and later home science environment were rare and relatively weak, failing to maintain after controlling for baseline home science environment.Interestingly, the most robust and longest lasting associations were observed in exploratory analyses differentiating home science activities from access to science-related materials. Indeed, home science activities, such as conducting experiments and visiting science museums, measured at 3 years as children were entering preschool, predicted scientific literacy through first grade. Home science materials, in contrast, failed to maintain any predictive power on their own (after controlling for baseline scientific literacy). One possibility is that materials are less important to emergent scientific literacy because children these young are unable to gain much from them of conceptual value through exploration on their own. Their value might therefore derive entirely from interactions they stimulate with parents, which would be subsumed by our measure of home science activities. It is also entirely possible that learning about the causal fundamentals of science at these early ages can be easily achieved with household items in the context of everyday activities like bathing and cooking, thus obviating the need for extensive collections of specifically science-themed materials.Although nuanced, and in need of replication and further confirmation through intervention studies, the overall pattern of results is consistent with enduring effects of the home science environment on the subsequent development of children’s scientific literacy. Moreover, this relation appears to be quite specific. Effects of science-related aspects of the home environment were generally stronger than those of cognitive stimulation more generally speaking, and some of the former maintained even when controlling for the latter. In addition, the home science environment differentially predicted scientific, in contrast to math or reading, literacy.In sum, this project demonstrates that children’s exposure to science related experiences as they enter preschool are predictive of their scientific literacy up to 4 years later. The fact that experiences at home appear to relatively decrease in importance as children spend more time in other contexts  is consistent with broad socio-ecological theories of development e.g., . Intereshttps://osf.io/z7cgd).The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found at: OSF  awarded to AB and CH.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The presence of multiple accessory spleens in the abdominal cavity is typically limited to two, with cases involving a higher number being exceedingly rare. Concurrently, accessory spleen infarction is remarkably uncommon, primarily resulting from torsion of the vascular pedicle. In this report, we present a case of a 19-year-old male who experienced infarction in one of four accessory spleens. Imaging diagnosis proved challenging, with the definitive diagnosis being made through postoperative pathology, revealing no torsion in the affected accessory spleen. Following surgery combined with anti-inflammatory and analgesic treatment, the patient exhibited an uneventful recovery. No complications were observed at the 3-month follow-up. This case indicates the challenge and difficulty of diagnosing accessory splenic infarction without torsion in imaging diagnosis. Employing a multimodality approach and diffusion-weighted imaging may aid in confirming the diagnosis. An accessory spleen is a congenital healthy organ comprising splenic tissue separated from the main body of the spleen. The size of the spleen varies from 0.5 cm to 2.8 cm in a maximal caliber, and only one accessory spleen is usually seen . AccessoA 19-year-old male patient complained of sharp pain in the left upper abdomen after heavy eating and drinking at a dinner. The pain persisted for 3 days, and was relieved after administration of anti-inflammatory medication and analgesia. Computed tomography (CT) of the lesion was performed on the 3rd day after the episode in a local hospital. Initially, a mass adjacent to the stomach, pancreas, and spleen, measuring 4.5 cm × 3.0 cm × 6.7 cm and with a well-defined margin, homogeneous soft tissue density, and edematous changes in the surrounding fat tissue, was revealed in an unenhanced CT image a. After 3 ; hemoglobin: 15.2 g/dL (14.0–18.0); platelet counts: 403 × 109 g/L (125–350 × 109); serum: AST 38 IU/L (15–40), ALT 35 IU/L (0–41); prothrombin time: 11.2 s (10.5–15); activated partial thromboplastin time: 33.2 s (22.7–31.8); D-dimer: 0.12 mg/mL (0–0.3); and CRP: 0.33 mg/dL (0.0–0.5). Tumor markers were within the normal range. On the 12th day after the onset, abdominal sonography revealed a large, oval-shaped, well-defined solid mass surrounded by hyperechoic mesenteric fat tissue in the upper left abdominal cavity. The entity demonstrated low inhomogeneous echogenicity compared to the spleen . Accessory spleens are most commonly found in the splenic hilum (75–90% of cases) and the tail of the pancreas. Accessory spleens are the most commonly encountered forms of developmental variations of the spleen . They ar%. AccessAccessory splenic infarction is a rare occurrence, and is mostly seen in children and young adults. It is rare 3.8%) in the elderly population (range: 14 days–75 years) % in the ,30,31,32In our case, no blood  system dHowever, a clear diagnosis of accessory splenic infarction was dependent on surgical pathology rather than diagnostic imaging prior to surgery, as in previously reported cases and in our patient. Visualization of a vascular pedicle originating in the splenic area was the most important element in the diagnosis of an accessory splenic infarction caused by torsion . HoweverThe imaging features of a normal accessory spleen, such as echogenicity, blood flow appearance on color Doppler, CT attenuation, iodine concentrations, signal intensity, ADC value, and enhancement pattern in CT and MRI were considered characteristic ,36,37,38In our case, fibrous tissue and slight vascular proliferation around the infarcted area were seen in microscopic specimens. The onset time and MRI examination time of these three cases were different, indicating that the change in the MRI signal was extremely sensitive to the period of infarction. The spleen is the organ with the most restricted diffusion in the upper abdomen . The ADCThe question of whether to perform a splenectomy for an infarcted accessory spleen has not been answered satisfactorily thus far. Involution or atrophy are the natural advancements of an infarcted accessory spleen . Early dIn this case report, we have identified several limitations that may impact the interpretation of our findings. First and foremost, our inability to definitively demonstrate the presence of a thrombus in the vessel feeding the infarcted accessory spleen due to a lack of direct intraoperative observation or pathological assessment contributes to the uncertainty of the actual cause. Additionally, while no torsion of the vascular pedicle was observed intraoperatively, it is important to consider that transient torsion might have been fixed spontaneously under general anesthesia, thus complicating our ability to exclude this potential cause of ischemia. Furthermore, the absence of sufficient pathological evidence for both transient torsion and thrombus formation led us to rely on a speculative approach to determine the most likely etiology of accessory splenic infarction in our case. Despite these limitations, we have made efforts to provide a balanced analysis of the potential causes, drawing support from the relevant literature and ensuring transparency in our discussion of the presented evidence.Accessory splenic infarction without torsion of the pedicle is extremely rare, and the imaging characteristics are complex due to their association with various periods of the infarction. However, these methods, such as ultrasound, CT, and MRI, can provide valuable insights into the disease and inform clinical decision making, thereby facilitating appropriate and timely intervention.


Dirofilaria. The major etiologic agent of canine dirofilariosis in the American continent is the zoonotic parasite Dirofilaria immitis. Existing reports of filarioid nematodes in Cuba are based solely on morphological and immunological analysis which do not allow unambiguous identification and/or direct detection of causal agents.Dirofilarioses are widespread diseases caused by mosquito-borne nematodes of the family Onchocercidae, genus cox1 gene fragments, the worms were unambiguously classified as D. immitis. Sequence analysis showed high identity of the gene fragments in this study with others previously obtained from D. immitis found in dogs, wolfs and jackals but also from mosquito vectors of D. immitis.Here we present the molecular characterization of filarioid nematodes found in a dog in Cuba. Based on the molecular and phylogenetic analysis of the 5.8S-ITS2-28S region and Dirofilaria in the country.Further studies are guarantee to better understand the epidemiological impact of canine dirofilariosis in Cuba as well as the competence of different species of culicid mosquitoes as vectors of The online version contains supplementary material available at 10.1186/s12917-023-03803-0. Dirofilaria, family Onchocercidae. Filariae of the Dirofilaria genus can infect wild and domestic animals of several orders including Rodentia, Artiodactyla, Perissodactyla, Carnivora, Lagomorpha, Edentata, and Primates [Dirofilaria worldwide [Dirofilariosis is a mosquito-borne disease with worldwide distribution caused by nematodes of the genus Primates . About 6orldwide .Dirofilaria repens and Dirofilaria immitis are two Dirofilaria species of special interest due to their negative impact on company animals  as well as their zoonotic potential [Canis lupus), foxes (Vulpes vulpes), and jackals (Canis aureus), but cats and weasels , can also be infected [D. immitis include respiratory distress, epistaxis, haemoptysis, ascites, exercise intolerance, and anorexia [D. immitis are asymptomatic or display no abnormalities in different laboratory tests [D. immitis with varying levels of sensitivity and specificity [otential , 4. The infected . Originainfected . Howeverinfected . Clinicaanorexia , 7. Howery tests . Severalcificity –11. Micrcificity , and thecificity  and conscificity .D. immitis [D. immitis [The diagnostic tests based on molecular methods have high sensitivity, and allow for filarioid differentiation , 15. In  immitis , 16, how immitis , 18.Dirofilaria including D. acutiuscula, D. striata, D. immitis and D. repens have been reported as causing infectation in dogs in the Americas [D. immitis is the most important causative agent of canine dirofilariosis in the continent [D. immitis in domestic dogs has been estimated to range from 1 to 12% [Several species of Americas –21, but ontinent . In Nort1 to 12% , while i1 to 12% , 23.Dirofilaria immitis was reported in Cuba in 1977 and in the 80’s several reports were published on the presence of this parasite in the country [D. immitis was reported in a dog using a coagglutination assay [D. immitis strains circulating in Cuba is available. Therefore, the aim of our study was to describe the morphological and molecular characterization of filarioid nematode collected from an infected dog in Cuba. Sequence and phylogenetic analyses showed to the presence of a D. immitis strain similar to that reported in other canids and mosquito vectors of D. immitis. country –27. In a country –27. In aon assay . HoweverDirofilaria. The dog had never been tested for heartworm or administered heartworm prophylactics.No clinical sign or symptoms indicative of diseases were observed in the dog. However, microscopic observation of Giemsa-stained blood smear revealed the presence of Microfilariae. The morphological examination of the microfilariae showed unsheathed microfilaria and structures such as the cephalic space, nerve ring, excretory pore, anal pore, and the terminal nucleus at the tail were identified Fig. , which a-ITS2-28S) rRNA region amplified a PCR amplicon at size corresponding to D. immitis DNA (542 bp) . Due to short fragment size, GenBank did not allow the deposition of the amplified cox1 fragment, which can be provided upon request.The pan-filarioid primer pair targeting the 5.8 S ribosomal RNA gene and internal transcribed spacer 2 , and Vulpes vulpes (Bulgaria MN596213). Analysed sequence showed also similarity to microfilariae of this species collected from insects of Ctenocephalides family (MW019916). Similarly, sequence of cox1 gene clustered with other D. immitis collected from canine, e.g., C. lupus familiaris , Vulpes zerda (USA MN945948); its primary vector Culex quinquefasciatus (Myanmar OL721654) and accidental host, e.g., human (Iran MH920260).Phylogenetic analysis of 5.8Sion Fig. A and coxent Fig. B placed D. immitis, has a wide distribution in Latin America [Dirofilariosis is an emerging parasitic infection of growing concern in the world . Canine  America , and the America , Curacao America , Grenada America , St Kitt America , Haiti [ America , Dominic America  and Puer America  and in i America . In Cuba America –28, and D. immitis, D. repens, and D. reconditum or Brugia malayi and Brugia pahangi), and requires considerable expertise [D. immitis in a dog from Cuba. Until now, no studies combining morphological and molecular diagnosis of canine microfilariae in Cuba have been published.The gold standard test for the diagnosis of microfilariae is the microscopic examination of blood smear stained with Giemsa or hematoxylin and eosin . Howeverxpertise . Here, w-ITS2-28S rRNA and cox1 gene fragments showed high identity with sequences obtained previously from D. immitis parasites found in dogs, wolfs and jackals [D. immitis obtained in the current study and other sequences uploaded to GenBank data BLAST is supported by previously published reports [Wolbachia sp., common bacterial endosymbiont of D. immitis influencing its survival and reproduction rate [Wolbachia-infected species [Sequence analysis of 5.8S jackals  as well  reports –42 and mion rate  but addi species .Dirofilaria infection in dogs, wild animals, and humans in Cuba. The role of different species and/or strains of culicid mosquitoes as vectors of D. immitis in the country should also be evaluated. These studies are important due to the risk of introduction of non-endemic Dirofilaria species or strains in the country. Additionally, veterinarians should be aware of the possibility that D. immitis infection might be an emerging condition with zoonotic potential in Cuba.Our findings justify a better characterization of the epidemiology of A one-year-old intact male mixed-breed dog born in Cuba and that had not left the country, underwent a routine examination by a local veterinary. The clinical inspection included anamnesis and clinical evaluation. The veterinarian performed a thorough physical examination that included several aspects such as body temperature recording, evaluation of mucous membranes, pulse assessment, hydration status, capillary refilling time, and manually checking the animal for skin lesions and tick infestation. Moreover, the evaluation involved observing the dog’s behavior, gait, and coordination, along with a neurologic examination the respiratory system was examined for symptoms. During the examination, the veterinarian searched for any signs or symptoms of dirofilariosis. The anamnesis was performed to gather information regarding the dog’s medical history, including previous illnesses, surgeries, and treatments. Any abnormalities or concerns detected were documented and addressed accordingly.A blood sample was aseptically drawn from the jugular vein using sterile Vacutainer needles and EDTA tubes  and maintained at 4 °C within 24 h of blood collection until further analysis.A thin blood smear was prepared, stained with Giemsa solution  and examined under a light microscope  at final magnifications of 100X and 1000X. Morphological identification was performed using keys previously reported .Total nucleic acid was extracted from 300 µL of EDTA-anticoagulated blood sample using the Wizard® Genomic DNA Purification kit , according to the manufacturer’s instructions. A negative control was set in which DNA extraction was carried out using 300 µL phosphate-buffered saline (PBS)  instead of blood. The quantitative and qualitative evaluation of nucleic acid extraction was determined using a Colibri Microvolume Spectrophotometer . The extracted nucleic acid sample was stored at 20 °C until further use.cox1) [cox1 gene fragment amplification consisted in a denaturing step at 95 °C for 60 s, 35 cycles of denaturing (20 s at 95 °C), annealing (20 s at 60 °C) and extension (40 s at 72 °C), a final extension (10 min at 72 °C) and a soak at 4 °C. An Eppendorf Mastercycler Nexus Gradient  was used for the PCR reactions. The molecular size of the PCR amplicons was examined on a 1.5% agarose gel for 5.8S-ITS2-28S rRNA and 2% agarose gel for cox1 gene, using a GeneRuler 100 bp Plus DNA Ladder (Thermo Scientific). Ethidium bromide was used as DNA-staining agent, visualized under UV light. Amplicon sequencing was commissioned to Eurofins MWG Operon .PCR reactions were carried out with species-specific primers to amplify fragments of 5.8S-ITS2-28S rRNA  and cytocox1)  . Next, sequences were aligned using MUSCLE algorithm available in MEGA X. Phylogenetic trees were constructed using maximum likelihood (ML) method and Tamura 3-parameter (T92) model, according to the lowest Bayesian Information Criterion (BIC) and Akaike information criterion corrected for small sample sizes (AICc) [In order to determine species identity and genetic diversity of filarioid nematodes collected in the current study, obtained sequences of 5.8Sre v.7.2 , analyzes (AICc) . ReliabiBelow is the link to the electronic supplementary material.Supplementary Material 1: Supplementary Figure


Bile acids (BAs), key regulators in the metabolic network, are not only involved in lipid digestion and absorption but also serve as potential therapeutic targets for metabolic disorders. Studies have shown that cardiac dysfunction is associated with abnormal BA metabolic pathways. As ligands for several nuclear receptors and membrane receptors, BAs systematically regulate the homeostasis of metabolism and participate in cardiovascular diseases (CVDs), such as myocardial infarction, diabetic cardiomyopathy, atherosclerosis, arrhythmia, and heart failure. However, the molecular mechanism by which BAs trigger CVDs remains controversial. Therefore, the regulation of BA signal transduction by modulating the synthesis and composition of BAs is an interesting and novel direction for potential therapies for CVDs. Here, we mainly summarized the metabolism of BAs and their role in cardiomyocytes and noncardiomyocytes in CVDs. Moreover, we comprehensively discussed the clinical prospects of BAs in CVDs and analyzed the clinical diagnostic and application value of BAs. The latest development prospects of BAs in the field of new drug development are also prospected. We aimed to elucidate the underlying mechanism of BAs treatment in CVDs, and the relationship between BAs and CVDs may provide new avenues for the prevention and treatment of these diseases. Cardio-metabolic disease (CMD) is a broad term describing cardiovascular disease (CVD) caused by systemic metabolic changes. Metabolic changes are mechanically involved in almost all forms of cardiovascular disease . CMD is Bile acids (BAs) are mainly synthesized in the liver and are the final product of cholesterol catabolism. Their components are cholesterol derivatives. Since the last century, evidence has shown that pathologically elevated BA circulation in liver disease is harmful to the heart , 4. In tBAs have not only been shown to play a key role in mediating oxidative stress, reactive oxygen species (ROS), mitochondrial dysfunction, cell membrane disruption and cellular damage ; more imIn this paper, we summarize the role of different BA synthesis pathways and several regulatory mechanisms on related cells in the development of CVDs, as well as their role in the development and treatment of CVDs and related metabolic diseases. In addition, we introduce the potential therapeutic effects of ursodeoxycholic acid (UDCA) and other BA derivatives.BAs are the product of cholesterol metabolism in the liver. BA metabolism is the main pathway for the human body to remove cholesterol, and BAs are also important substances because they affect lipid absorption. Based on their structure, BAs can be divided into primary and secondary BAs. BAs can be further classified into bound and free BAs according to whether they are combined with glycine or taurine .Currently, the major BAs identified in humans include chenodeoxycholic acid (CDCA), cholic acid (CA), and a small amount of lithocholic acid (LCA). UDCA and muricholic acid (MCA) are primary BAs in rodents rather than in humans. According to the different groups in the R1/R2/R3/X position, the lipophilicity of BAs is different. BAs are formed in the liver through a complex process and finally stored in the gallbladder. The formation process includes multiple reaction steps involving at least 17 different enzymes .There are two ways to synthesize BAs, namely, the classical pathway and the alternative pathway . In the enterohepatic circulation ..11].According to previous studies, increased hydrophobic BA serum levels are associated with various metabolism-related diseases .2+-activated potassium (K+) (BK) channels  through S1P1 receptor-mediated hypoxia ) has been synthesized with CDCA. OCA is currently the most clinically advanced BA derivative. It is anticipated that OCA can be used as a new therapeutic drug to replace UDCA. OCA has a variety of biological and pharmacological applications. As a semisynthetic BA analog, it has a strong binding affinity to FXR . Wu et aet al.. confirmet al..The metabolic effects of several drugs commonly used in the clinical treatment of diabetes are generally dependent on the regulation of BA metabolism. Among them, metformin is thought to have a hypoglycemic effect by reducing the intestinal absorption of BA . The synSeveral studies have shown that hydrophobic BAs can be blocked by inhibiting ER stress, free cholesterol-induced cell death in macrophages, and the presentation of major histocompatibility complex (MHC)-related antigens. Fat production increases the risk of developing athero-sclerosis and other cardiovascular diseases . It has FXR has a tissue-specific role in the development and progression of atherosclerosis, and its prevention depends not only on hepatic FXR activation but also on global and intestinal depletion of FXR -117. Actin vivo experiments to determine the BA-FXR interaction. LXR has also been reported to regulate CVDs such as atherosclerosis. Bradley et al. showed that activation of LXR reduces the formation of atherosclerotic lesions I [UDCA was shown to protect CMs against arrhythmias mediated by adenosine triphosphate-gated K [Ca2+]I . Other sIn patients with chronic heart failure, the serum concentration of secondary BAs was found to have increased, thus resulting in a larger proportion of secondary BAs in the BA pool . BAs actIt has been reported that TGR5 agonists enhance the adaptability of the heart to physiological, muscle strength, and hemodynamic stress, further inducing changes in its protective mechanisms . TherefoThe deterioration of cholestatic disease leads to liver cirrhosis . The couIn models of liver cirrhosis, several mechanisms associated with BAs have been proposed to induce vasodilation of splanchnic and systemic vessels, resulting in hyperdynamic circulation. Therefore, research on the relationship between the pathophysiological characteristics of cirrhosis cardiomyopathy and the abnormal metabolism of BA has sparked interest among researchers . AccordiTGR5 can regulate metabolic homeostasis in the heart. Activation of the TGR5 signaling pathway can prevent nonalcoholic steatohepatitis by reducing vascular and liver damage . S1P1R aIn recent years, UDCA, a highly hydrophilic secondary BA, has been shown to act as an alternative drug to protect hepatocytes by promoting bile flow and reducing liver enzyme levels . FurtherMuch of the abovementioned evidence suggests that BAs can affect or regulate the function of the heart in cirrhotic cardiomyopathy. However, to date, there have been few clinical studies specifically targeting the interaction of BAs and cardiovascular function in patients with liver cirrhosis, and more trials are needed in the future.As the study progressed, researchers have begun to explore the diagnostic and prognostic value, as well as pharmacological application, of BA metabolism and related signaling pathways in cardiovascular and metabolic diseases.et al [www.ncbi.nlm.nih.gov/geo), we analyzed the expression level of the regulatory genes and receptors related to BA synthesis and metabolism in dilated cardiomyopathy (DCM) patients, hypertrophic cardiomyopathy (HCM) patients and nonfailing healthy donors. According to the obtained table and heatmap, the expression of most of the BA metabolism-related genes was disturbed compared with that in the nonfailing healthy donors , colesevelam is approved for the treatment of type 2 diabetes mellitus (T2DM) and hyperlipidemia . Colesevet al. [et al. [Chevli PA et al.  investig [et al.  showed t [et al. . However [et al. . These fIn a tissue Doppler imaging study, the authors investigated the association of TBA levels and fetal cardiac function in women with intrahepatic cholestasis (ICP). Elevated maternal and fetal serum BA levels in severe ICP have been found to be associated with abnormal fetal cardiac phenotype and fetal cardiac insufficiency compared with those in healthy women with normal TBA levels and in women with mild ICP. In particular, when the maternal TBA level was greater than 440 mmol/L, the incidence of fetal complications, such as spontaneous preterm deliveries, asphyxia events, and meconium staining, was significantly higher . Howeveret al.[In a prospective, single-center study, elevated levels of specific secondary BAs and decreased levels of primary BAs were found in patients with chronic HF. Specifically, Mayerhofer CCK et al. measuredet al. showed tin vitro and in vivo studies in humans have shown that TMAO has pleiotropic negative effects on the cardiovascular system [Furthermore, BA homeostasis is jointly maintained by hepatic and intestinal BA signaling pathways. BA induces enterohepatic feedback signals by releasing intestinal hormones and regulates enterohepatic circulation . The rolr system -168. TMAr system , 170. Ser system , 172. Ser system , 174. ThIn addition, in recent years, an increasing number of studies have confirmed the pharmacological applications of bile acid derivatives. BAs are considered to be very helpful for the preparation of novel drugs due to their rigid backbone and potential for surface amphiphilicity . The broBAs serve as an attractive cornerstone for designing novel hydrogel systems for the delivery of biomolecules, drugs and vaccines , 178. ThRecently, evidence has accumulated indicating that the relationship between BA metabolism disturbances and CVDs is closely related. When BA metabolism is disordered, a series of cardiac dysfunction and CVDs may also be present. In this paper, we have clarified the metabolic mechanism of BAs and their pharmacological potential in regulating cardiovascular function. BA signaling plays an important role in different cell types through receptor-dependent or channel-mediated mechanisms. Future work should aim to further elucidate the deeper interactions between BAs and their receptors to facilitate the development of new treatments for CVDs.Current clinical studies as well as our previous metabolomic and bioinformatics analyses have revealed that TBA levels, BA pool composition ratios, and BA-related receptors are partially disturbed in human CVD. However, whether altered BA in humans can serve as a potential biomarker in the pathogenesis of CVD remains unclear and warrants further study.Fortunately, UDCA has now been found to play a protective role in CVD, although its specific protective mechanism has not been fully elucidated. Currently, some drugs targeting UDCA and its alternatives, some synthetic BA analogs such as OCA and Colesevelam, have been used in clinical practice. However, to further confirm their importance in cardioprotection, more information on their application in preclinical and clinical studies should be provided in the future.Recent reports have focused on the application of BAs in the preparation of new drugs. BAs have become the main molecules in drug carrier systems due to their good compatibility with different biologically active compounds, showing great potential in medical and biological applications. However, there is no further research on the development of drugs for the treatment of CVDs. Furthermore, the safety and effectiveness of drugs targeting BAs should be evaluated in the treatment of patients with diverse CVDs. Therefore, it is expected that the uniqueness of BAs as drug carriers can be fully utilized to complete further development in the future.In addition, it is necessary to conduct studies to investigate the effects of drugs that modulate BA metabolism or signaling pathways on lipid metabolism and other related proteins in patients with CVD in the future. This can form a better pharmacological basis for the clinical treatment of CVDs such as atherosclerosis, coronary heart disease and heart failure.Another potentially interesting area of research is the possible role of VD as a therapeutic target for cardio-vascular diseases. There is evidence that VD may affect cardiovascular function through multiple pathways and that VDR plays an important role in BA transport, metabolism, and detoxification. However, current research on the association between BA and its receptor, VDR, is still insufficient. More research in this area will be needed in the future to further elucidate whether altered VD may serve as a potential biomarker in cardiovascular pathogenesis.It is worth noting that current research on the mechanism of BA metabolism and CVD progression is mainly carried out in rodents. Nevertheless, their BA metabolism is fundamentally different from that of humans. Some animals do not have gallbladders, such as rats. Therefore, some preclinical findings may vary by species, and the use of animal models to study BA metabolism still has some limitations. Translating findings from animal models into humans is challenging. Future studies need to consider important issues related to species limitations in clinical trial design and seek more efficient ways to explore the potential roles of BA metabolism and BA pool components in human CVD clinically.Furthermore, the manipulation of factors affecting BA metabolism is unclear. Further research needs to consider whether some factors, including gender dimorphism of BAs and potential signaling crosstalk between gut microbiota and BA signaling pathways, can alter clinical outcomes. Further studies have the potential to open a new era for the application of BA in clinical practice to further prevent the risk of CVD.