Secondary lung cancer (SLC) can offset the benefit of adjuvant breast radiotherapy (RT), and risks compound sharply after 25 to 30 years. We hypothesized that SLC risk is mainly an issue for early-stage breast cancer, and that lives could be saved using different RT techniques.The SEER database was used to extract breast patient age, stage survival, and radiotherapy utilization over time and per stage and to assess the factors associated with increased SLC risk with a multivariable competing risk Cox model. The number of SLC was calculated using the BEIR model modified with patient survival, age, and use of RT from the SEER database. Stage distribution and number of new breast cancer cases were obtained from the NAACCR. Mean lung dose for various irradiation techniques was obtained from measurement or literature.Out of the 765,697 non-metastatic breast cancers in the SEER database from 1988 to 2012, 49.8% received RT. RT significantly increased the SLC risk for longer follow-up (HR=1.58), early stage including DCIS, stage I and IIA (HR = 1.11), and younger age (HR=1.061) . More advanced stages did not have significantly increased risk. In 2019, 104,743 early-stage breast patients received radiotherapy, and an estimated 3,413 will develop SLC (3.25%) leading to an excess of 2,900 deaths (2.77%). VMAT would reduce this mortality by 9.9%, hypofractionation 26 Gy in five fractions by 38.8%, a prone technique by 70.3%, 3D-CRT APBI by 43.3%, HDR brachytherapy by 71.1%, LDR by 80.7%, and robotic 4π APBI by 85.2%.SLC after breast RT remains a clinically significant issue for early-stage breast cancers. This mortality could be significantly reduced using a prone technique or APBI. With the generalization of mammography screening, breast cancer can be diagnosed at an early stage , and theCardiac morbidity appears relatively soon after the radiation treatment, generally 5 to 10 years following exposure of the heart . It is wIt is difficult to use LAR in making the decision to adopt a new radiation technique, or discussing the radiation treatment with a given patient, since this risk assumes the patient would survive for a long time. It is also challenging to get a clear picture from population-based studies of the absolute number of secondary lung cancers for patients diagnosed with breast cancer today. Most meta-analysis or registry studies calculate the risk of secondary lung cancer based on patients treated a long time ago , 10, 12.This study aimed first at, confirming that early-stage breast cancers have a higher risk of secondary lung cancer; second, comparing the true number of secondary lung cancers for different radiotherapy techniques; and third, estimating the number of lives that could be spared depending on the technique for early breast cancer patients diagnosed today.The SEER*Stats software version 8.3.4 was used to extract a case-listing from the Surveillance Epidemiology and End Results (SEER) 18 Custom Database with additional treatment information, which includes radiotherapy delivery information . This dabeam radiotherapy”, or “combination of beam with implanted radioisotope”, or “radiotherapy delivered but method not specified”, or “radiotherapy recommended but it is unknown if administrated”. Conversely, patients were deemed not to have received radiotherapy when the record indicated “no/unknown” or “radiotherapy recommended but refused”. The occurrence and date of lung cancer after breast cancer was matched using the patient’s SEER ID.The collected information included the patient’s SEER ID, tumor site, cancer stage, age at diagnosis, month and year of diagnosis, survival in months, vital status at study cut-off, and delivery and type of radiotherapy if any. When patients had multiple breast cancer records, it is either the first one treated with radiotherapy or, if radiotherapy was not delivered, the first one diagnosed that was chosen as date of diagnosis. Patients were deemed to have received radiotherapy when the record indicated “The final database was stratified in prognostic groups of relatively similar sizes, including DCIS, T1a N0, T1b N0, T1c N0, Stage IIA T2 tumors, Stage IIA N1 tumors, Stage IIB, Stage IIIA, Stage IIIB, or Stage IIIC. Radiotherapy utilization was evaluated per decade for each prognostic group, ranging from 1988 to 1997, 1998 to 2007, and 2008 to 2012.The overall survival and lung cancer–free survival were calculated for each patient stage group using Kaplan Meyer statistics. Univariate regression analysis was used to identify the risk factors of developing secondary lung cancer. A multivariable competing risk Cox proportional hazard model was developed using a stepwise regression (PROC PHREG procedure in SAS) to account for competing risks of all-cause mortality. Independent variables included the breast cancer stage, age at diagnosis, year of diagnosis, overall survival, and the delivery of radiotherapy, as well as the resultant interaction terms between these variables. Statistical analyses were performed using SAS/STAT 14.2  or SPSS 25.0.0.1 . Because multiple tests were performed, the level of significance had been set to p<0.001.To calculate the number of patients at risk of developing a secondary cancer after breast radiotherapy, a modified BEIR VII model for a female breast cancer patient was constructed, accounting for the age and stage distribution at diagnosis, present use of radiotherapy, and survival per stage derived from the SEER cohort , 17. StaA total of 900,085 patients with 1,079,406 cancer records were found in the SEER database. After removing stage IV, unknown stages, unknown dates of event, and patients with confounding factors, the cohort included 765,697 patients. The median age was 60 years, with an interquartile range of 21 years. Overall, 15.2% of patients had two cancer records, 1.9% had three records, and 0.24% had more than three. Breast cancer was the most frequent new cancer event recorded.in situ breast disease is not appropriately reported to the SEER. To compensate for this potential under-reporting, the final calculation of lung cancer risk and lives saved used the 2012–2016 North American Association of Central Cancer Registries (NAACCR) ((NAACCR)  stage diversus 50.2%, respectively. The usage of radiotherapy changed over time, with a significant increase in the most recent cohorts. While there was 40.3% of patients receiving adjuvant radiotherapy during the 1988–1997 decade, this increased the following decades, 52.0% between 1998 and 2007 and 52.2% between 2008 and 2012 (p<0.001). Of note, the radiotherapy usage decreased from 65 to 53% for the very favorable T1a N0 stage group.A total of 13,689 lung cancers were detected in the cohort. Using a stepwise regression procedure, all factors were highly significantly associated (p<0.001) with an increased risk of developing secondary lung cancer, including the use of radiotherapy, an earlier cancer stage, a younger age, an earlier year of diagnosis, and a longer survival. The interaction factors were also highly significantly correlated (p<0.001). Accounting for the interaction factors, the Hazard Ratio (HR) to develop a secondary cancer was higher for patients receiving radiotherapy and treated between 1988 and 1993 , compared to those receiving radiotherapy between 1994 and 1999  and those receiving radiotherapy between 2000 and 2005 .Out of the 268,600 women diagnosed with breast cancer in the USA in 2019, an estimated 205,318 had an early stage including DCIS, stage I, and node negative stage IIA (We previously reported that using a VMAT technique optimizing the lung protection could slightly reduce the mean lung dose, eventually saving 9.9% of those excess lung cancer deaths. Using a lower total dose, 26 Gy in five fractions, would reduce the mean lung dose and eventually reduce by 38.8% the excess lung cancer mortality. Based on a literature review, Aznar reported that the prone technique enables a significant reduction of the lung exposure with a mean lung dose of about 0.6 Gy . This woAPBI also induces a lower mean lung dose, though not all techniques are equal. We measured for external beam 3D-CRT APBI a mean lung dose of 1.146 Gy, which would reduce the excess mortality by 43.3%, and for multicatheter HDR brachytherapy a mean lung dose of 0.584 Gy, enabling the reduction of 71.1% of the lung cancer death . Using lAccounting for changes in radiation usage, protocols and techniques, current survival outcome, and patient’s stage and age characteristics, this study confirms the long-term risk of secondary lung cancer after radiotherapy for breast cancer patients diagnosed today. It adds to knowledge that the issue is statistically significant mainly for early-stage and young patients, who have a higher probability to have 20 to 30 years of survival , 14. ThiAlthough they occur late in the patient’s life, the absolute risk of excess lung cancers and deaths calculated for good prognosis cancers in the present work is high and in the same order of magnitude as radiation-induced cardiac morbidity and mortality. So, the same weight should be placed on reducing the mean lung dose when planning breast radiotherapy, as it is to minimize the mean heart dose . The gooAPBI is recommended by various societies for early breast cancer stages, which our study also shows to be the main benefactors for the secondary lung cancer prevention –35. In 2The FAST-Forward regimen has been strongly recommended by experts to minimize travel and potential exposure of frail patients at the hospital , 37. As Caution is needed in interpreting clinical outcomes from modeling, but similarly to the evaluation of cardiac morbidity, this might be the only possible strategy to assess very long-term breast radiotherapy toxicities . It is nWe used several adjustments in our model. The finding that DCIS might be underreported into the SEER database led us to opt for a hybrid model using the survival and radiotherapy utilization per stage from the SEER database, and the stage distribution from the NAACCR . The calCaution is also needed when applying the excess lung cancer risks reported in this work to patients with different anatomy, like a smaller or a larger breast. On one hand a larger-breast patient may experience higher dose to the lung due to the larger radiation scattering volume. On the other hand, a larger body mass index means more shielding tissue and therefore less dose scattered to organs as shown on Woo’s prospective study measuring scatter dose with skin dosimeters . This meIn conclusion, this study confirms that, accounting for the current utilization and techniques of radiotherapy, patient’s characteristics, and outcomes, a significant number of patients diagnosed with early breast cancer will succumb after two to three decades to radiation-induced secondary lung cancer. This could be prevented by reducing the mean lung dose with techniques like the prone technique, brachytherapy, or ultra-hypofractionated 4π robotic APBI.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.J-PP, HD, and DW built the model and did the analysis. J-PP and NH collected experimentally the mean lung doses for the model. J-PP, NH, HD, MN, and FV wrote the first draft of the manuscript. J-PP, NH, HD, DW, MN, and FV finalized the manuscript. All authors contributed to the article and approved the submitted version.Author FV is employed by 21st Century Oncology, Inc.The remaining 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.


Cognitive impairments, such as steep delay discounting, have been correlated with substance-related disorders. However, antisocial traits, cognitive inflexibility, and loss discounting have been barely considered despite having a high relationship with problematic consumption. This study aims to identify the predictive power of these variables in four types of drug use. Fifty-two adolescents (age range of 13 to 19 years) were assessed with a substance involvement test, four discounting tasks using $3,000, a card sorting test, and antisocial screening. Discriminant analysis with simultaneous estimation and varimax rotation was carried out. Function one included discounting of both losses, function two AT and CI, and function three probabilistic gains. The three functions explained 60.1% of the variance. The results show that preference for small and soon punishments and larger and unlikely punishments distinguished non-use and experimental use of moderate consumption and problematic consumption. High antisocial traits and low cognitive inflexibility distinguished experimental use groups of non-use. Risk-taking did not discriminate effectively between moderate consumption and problematic consumption. A replication of this study with a larger sample size is recommended to verify the results. Substance-related disorder causes significant negative health consequences  and involves expensive healthcare and criminal justice costs Gibbons, . SeveralDefined as persons aged 10 to 19, adolescents are at a high level of risk for drug use involvement  have demonstrated the prediction of both onset use and frequency of use in persons without SRD, with odd ratios between 1.34 and 2.4, as well as explained variance between 9.1 and 43%  have demonstrated their potential for prediction  and five predictive factors . The second objective of this study is to evaluate how these variables contribute to predicting each type of involvement. This research goal is important given that only few studies have included these variables in the same study. This inclusion may facilitate the identification of interaction effects, resulting in predictions that are more accurate . Additionally, all the participants provided the written informed consent following the Declaration of Helsinki, and they were not compensated with money for their participation.Mini International Neuropsychiatric Interview. A structured interview with dichotomy answers based on DSM-IV and ICD-10 was used. Each affirmative response was scored with one point. A score greater than three indicates a possible episode or disorder. The interview has test–retest reliability ≥ 0.75. The psychotic disorder section includes 10 questions, such as “Have you ever heard things other people couldn't hear, such as voices?” The (hypo-)manic episode section has nine questions such as “Are you currently feeling ‘up’ or ‘high’ or ‘hyper’ or full of energy?”  for cognitive impairment detection. The MoCA assesses executive function, attention, abstraction, working memory, language, orientation, and visuospatial abilities. A score below 19 points indicates impairment. It has shown a Cronbach's α of 0.88 in Spanish-speaking Hispanics and 0.89 in the Mexican population, as well as convergent validity of 0.83 . The ASSIST is a test developed for the WHO. It consists of eight questions about lifetime use, consumption, and consumption troubles related to the last 3 months. For example, “During the past 3 months, how often have you failed to do what was normally expected of you because of your use of (drug)?” The test includes questions about the use of alcohol, tobacco, cannabis, amphetamines, opioids, sedatives, cocaine, hallucinogens, and inhalants. Each substance is scored from questions two to seven. This test has shown an internal consistency of 0.8 and 0.87  Platform SE b version 7, for Windows®7 and 8] were applied: delay gains, delay losses, probability gains, and probability losses. Each of them consisted of a block of four practice trials and a block of 25 trials. The initial value of the large alternative was 3,000 Mexican pesos. The delays used were 1 week, 1 month, 6 months, 1 year, and 3 years. The probabilities were 10, 25, 50, 75, and 90%. All delay and probability discounting tasks used an adjusting-amount procedure that converges on the amount of an immediate certain outcome equal in subjective value to a delayed or probabilistic outcome  includes 64 cards with figures. Each card varies in color , number of figures , and shape . A sheet with four cards is placed on top of a desk, and 64 cards are taken one by one. Each time, the participants must sort the card according to one of the three possible criteria and categorize it with one of the four cards presented on the desk. The evaluator indicates when an answer is correct or incorrect. The analysis units of both tests were perseverative errors and deferred perseveration. This test has a reliability of 0.8 in the Mexican population , impulsivity , and narcissism . It has an internal consistency of 0.79 in Mexican adolescents manic episode evaluation, and six because of not answering the ASSIST.The participants were assigned to a group based on their ASSIST scores. The non-substance-use group was composed of participants who scored zero in all items of question one (never used any drug). The experimental substance-use group was composed of participants who scored three points or more in question one (tried drugs at least once in your life) but zero points in question two (no use in the last 3 months). The moderate substance-use group included participants who acquired low-risk scores, an alcohol score between 1 and 10, and other substances between 1 and 3. The problematic-use group consisted of participants who attained moderate risk scores, an alcohol score between 11 and 26, and other substances between 4 and 26.Due to the coronavirus disease-2019 contingency, the assessments were suspended, and the researchers were unable to reach the sample size. Despite this setback, the obtained sample size met the minimum requirement for discriminant analysis: a minimum of five observations per variable and the group size being equal to or greater than the number of variables plus one  was used  = 32.22, p = 0.367]. Hence, mean group imputation was carried out for variables with <10% of missing data, and regression imputation for the group for variables with more than 10%. Normality criteria for small samples were employed: Shapiro–Wilks no significant, z skewness, and z kurtosis smaller than 1.96 as well as scores not exceeding 2.5 standard deviations  was made with variables that show means statistically different. The effect size was calculated considering ω2 values: small between 0.01 and 0.05, a medium between 0.06 and 0.13, and large ≥0.14  post hoc was estimated using G*Power, considering 0.8 an adequate power . The education year mean was 9.8 (SD = 1.7). On average, the adolescents with employment have worked 16.14 months (SD = 7.62). Furthermore, the education mean of the mother was 11.55 (SD = 2.4), and the education mean of the father was 11.83 (SD = 2.72). Additionally, there were no significant differences among the groups concerning sociodemographic characteristics, cognitive functioning, psychopathological symptoms, and family background  were enough to carry out a discriminant analysis. Moreover, the mean of four variables differs significantly between groups with medium to large effect size and low statistical power: probabilistic gains , delayed losses , probabilistic losses , and antisocial traits . Although post hoc only confirmed differences in probabilistic gains  and probabilistic losses . In delayed losses, the closest comparison to significance was between moderate use and experimental use , whereas antisocial traits were associated with problematic use   see .Three functions result from discriminant analysis, together explaining 60.1% of the dependent variable variance. In the first function, delayed losses and probabilistic losses contributed the most, whereas, in the second function, antisocial traits and deferred perseveration contributed the most. Moreover, perseverative errors were very close to 0.4 in the second function. However, its value on the first two functions was very similar; thus, it is not clear to which it belongs. In the third function, only probabilistic gains had a substantial contribution to prediction. Regarding general contribution, probabilistic losses and delayed losses had the highest potency index. These were followed by probabilistic gains, antisocial traits, deferred perseveration, and perseverative errors, which had similar inputs. As for delayed gains, it had a minimal contribution as individually as overall with absolute values.p < 0.01, with non-use and problematic use groups being the most correctly classified  were above the classification thresholds. Likewise, the Q value of Press was higher than the critical value (6.63) see . Therefofied see . Additiop < 0.01.Prediction accuracy decreased in all levels, except in the moderate-use group where the percentage was the same. Nonetheless, the hit ratios were kept above analysis estimation thresholds, except for the problematic-use group located below the maximum chance criterion by 0.9%. Moreover, the Q value of Press also kept higher than the critical value see . Hence, In the non-use group, three cases were misclassified, from which only two had a higher probability of being classified in the wrong group in contrast with the likelihood of the correct one. In the experimental use group, all the individuals had a higher probability of being misclassified. In the moderate and problematic consumer groups, the cases had almost a double or more than double likelihood of being assigned to the wrong group. Furthermore, in the moderate-use group, most of the participants were classified as non-use. Thus, two analyses were carried out. The first analysis grouped all the misclassified cases together. The second analysis examined the moderate-use groups, differentiating between those classified as non-use and those with other classification.U = 0, p = 0.01, r = −0.18) and smaller AUC in delayed losses  than correctly classified, both with a small effect size. In the experimental-use group, misclassified cases had less antisocial traits than correctly classified cases, with a large effect size . In the moderate-use group, misclassified cases showed larger AUC in probabilistic gains  and in delayed losses , as well as more deferred perseveration . Lastly, in the problematic-use group, significant differences were not found.In the non-use group, misclassified cases showed larger AUC in probabilistic gains (H(2) = 7.05, p = 0.017], delayed losses , and antisocial traits . Post hoc with Bonferroni correction  revealed that misclassified 2 had larger AUC in probabilistic gains than correctly classified with small effect size . In turn, misclassified 1 showed lower antisocial traits than correctly classified with small effect size ; it was also smaller than misclassified 2, albeit it does not reach enough significance . Regarding delayed losses, correctly classified had smaller AUC than misclassified 1  and misclassified 2 ; however, these comparisons did not reach required significance.The analysis with only moderate substance use distinguished those misclassified values as non-use (misclassified 1) and others (misclassified 2). There were significant differences among correctly classified and two misclassified groups in probabilistic gains [This study aimed to identify the predictive power of gain and loss discounting, cognitive inflexibility, and antisocial traits in four types of drug use. The characteristics of the sample used were adequate. All the groups were equal in terms of sociodemographic data, global cognitive functioning, psychopathological symptoms, and family background. In comparison to other research studying adolescent drug use, the participants presented comparable drug use trends   than the problematic consumer group. The first finding supports other studies that have also found that groups with heavy and chronic substance use show more antisocial behavior than the non-substance use group  and presented less risk-taking than the problematic-use group . As for delayed losses, research in adults has found similar results. Those with higher alcohol and marijuana consumption exhibit higher discounting of delayed losses than those without use  in the third function, the analysis did not show accurate discrimination among the groups. This finding supports similar studies where risk-taking was not found to be a significant predictor of substance use  and delay discounting of negative outcomes , indicating these adolescents may display their riskiest drug involvement at a future time point or during adulthood  showed higher rates of depression (depression symptomology or indication of major depressive episode) but low antisocial behavior, whereas the misclassified case had a large AUC in delayed losses and low AT. The second trajectory (with an increase of tobacco use into adulthood) was marked with high rates of antisocial behavior, as well as low risk-taking and depression (same as above), likewise correctly classified: high APSD score, aside from small AUC in probabilistic gains and delayed losses. The third trajectory (with an increase in marijuana use in adulthood) was marked with high risk-taking behavior and depression  due to damage from chronic and heavy substance consumption . Hence, it is necessary to look for interventions that achieve a high level of awareness among adolescents regarding drug-related damages, such as SRD, and potential negative consequences. It is important not to underestimate the magnitude of a potential consequence related to drug use, especially in consideration of the high-risk probabilities demonstrated by the different drug use trajectories. Further, these findings demonstrated the relevance of antisocial traits, even for experimental consumption. Therefore, an intervention that diminishes antisocial traits could prevent adolescents from taking drugs. It is important for such intervention programming to consider the different drug use trajectories and how maturation-related changes can account for the discounting of delayed gains. Therefore, it would be further necessary to identify specific factors that distinguish moderate substance use from problematic substance use and to consider additional variables, such as depression and anxiety. These efforts could help to precisely identify which variables are active during the development of SRD. That said, it is important to note that this study is limited in that the researchers gathered a large sample size and yet received a low statistical power. It is suggested that the study be replicated with a large sample.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Institutional Review Board of the Sonora Institute of Technology (ID 37). Written informed consent to participate in this study was provided by the participants'alegal guardian/next of kin.LH and DM designed the study and wrote the protocol. LH conducted literature searches and provided summaries of previous research studies. DM wrote the first draft of the manuscript. LA-C conducted the statistical analysis. All authors contributed to and have approved the final manuscript.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 basic function of toothpastes is biofilm removal in order to prevent caries and gingivitis. Toothpastes should provide maximal fluoride availability, optimal abrasivity, and ingredients that do not interfere with fluoride release but should have additional beneficial effects. Further, the effect on cells of the oral cavity is of the utmost importance. We investigated several biological parameters of a new toothpaste (AirFlow-AF) that contains fluoride, xylitol and erythritol but no sodium lauryl sulfate and compared them to commercially available toothpastes . The half lethal concentration (LC50) as well as the proliferation behavior on gingival (GF), periodontal ligament (PDL), and mouse fibroblast cells (L929) were was tested. The mean LC50 values of AF on GF, PDL, and L929 were 16.2, 10.9, and 9.3, respectively. In comparison, the four other toothpastes showed mean LC50 values of 1.5 (OB), 1.2 (OM), 1.4 (Se), and 27.7 (Ze) on GF. Mean LC50 values on PDL and L929 were 1.0 and 0.2 (OB), 3.7 and 0.9 (OM), 1.2 and 0.6 (Se), and 25.4 and 5.6 (Ze), respectively. Proliferation behavior mainly confirmed the LC50 values. While cells after stimulation with AF returned to almost unimpaired proliferation behavior at 6%, cells were still strongly impaired after stimulation with all tested commercially toothpastes. AF showed high biocompatibility with different cell types. Caries and periodontal diseases still exist, negatively influencing both individual well-being and the general economy . AlthougIn addition to fluoride, there are other naturally occurring substances that may be effective against the risk of tooth decay. Examples of these are erythritol and xylitol. Erythritol is a naturally occurring sugar alcohol that is approved as a sugar substitute with no or minimal laxative effect and without any quantity restriction as a food additive. In various in vitro and clinical studies, erythritol showed an inhibitory effect on biofilm formation, on the presence of caries-inducing bacteria, and on the susceptibility of subjects to caries ,4,5. XylIn contrast to fluoride, sodium lauryl sulfate (SLS) is an ingredient that should be viewed with some caution. SLS belongs to the group of surfactants and its foaming effect ensures good distribution of the toothpaste in the oral cavity. In addition, surfactants reduce the surface tension due to their simultaneous hydrophobic and hydrophilic properties, which gives them a cleansing effect. Although SLS is one of the less expensive surfactants and is still widely used, some toothpaste manufacturers are already abandoning SLS as a surfactant because it is considered problematic for the cells of the oral mucosa in some in vitro studies. Furthermore, clinical reports on the association between more frequent occurrence and longer persistence of ulcerative aphthae and the use of SLS-containing toothpastes reinforce the use of alternative surfactants ,9,10.What we already know is that toothpastes that contain certain ingredients for foaming have negative effects on cells in vitro and also on the oral mucosa in vivo ,12,13. Fw/v%) and dissolved in sealable plastic tubes with a magnetic stirrer bar at 350 rpm. Afterwards, centrifugation at 16,000× g for 10 minutes of the slurries was performed and the toothpaste-conditioned medium (TCM) was filter-sterilized [w/v% toothpaste in serum-free medium.The toothpastes used for the experiments were AirFlow-AF , Zendium Complete Protection-Ze , Sensodyne Repair-Se, Odol Med3 Classic-OM, and Oral-B Repair-OB . Information on the amount of fluoride added and the surfactant used is summarized in erilized ,12. For The pH of each toothpaste was measured at room temperature using a pH meter  after mixing 20 g toothpaste and 40 g artificial saliva with an IKA Ultra-Turrax T 25 digital Dispenser for 10 s.The free fluoride was measured using a fluoride ion-specific electrode . To do so, paste slurries as above were prepared and homogenized by agitation for 1 h and the fluoride ion concentration was calculated and expressed in parts per million (ppm). The calibration curve in the expected fluoride range was performed. All measurements were made in triplicate.2 and 95% humidity. Fibroblasts that grew out from the explants were used for the experiments up to the seventh passage. For the respective experiments, the three types of cells were each seeded in microtiter plates  at 30,000 cells/cm2 one day before incubation with the different toothpastes.A murine cell line (L929) often used to perform cytotoxicity tests and human cells of the oral cavity exposed to toothpaste during oral hygiene  were used to evaluate the half lethal concentration of the toothpastes. Furthermore, the proliferation behavior of the cells after incubation with the corresponding toothpaste slurries was investigated. Human gingiva and periodontal ligament fibroblasts were obtained from tissue grafts after approval by the local ethics committee (123/2020 SFU) and after informed consent by the donors. To isolate the cells, tissue explants from the periodont and the gingiva of extracted wisdom teeth were cultivated in Dulbecco’s Modified Eagle Medium  supplemented with 10% fetal calf serum  and antibiotics (Invitrogen) at 37 °C, 5% CO2 into 96-well plate culture dishes , were incubated with different concentrations  of freshly prepared TCM. After two minutes, cells were washed with phosphate-buffered saline (PBS), which was replaced by serum-free media containing MTT . Optical density of formazan crystals, formed by NAD(P)H-dependent oxidoreductases and dissolved in dimethyl sulfoxide, was measured after incubation for 2 hours at 37 °C using a microplate reader . The results of the viability tests were normalized to untreated cells (set at 100% viability) and the half lethal concentration (LC50) was calculated by an exponential regression analysis using the formula y = m × e^(b × x)  [L929 cells, gingival fibroblasts (GF), and periodontal ligament cells (PDL), all seeded in growth medium at 30,000 cells/cm0: LC50) ,12.2 and stimulated the next day. For stimulation with the respective toothpastes, L929 cells were incubated with different concentrations of TCM for exactly two minutes, since two minutes is the recommended and also the average time for toothbrushing [For analyses of the proliferation behavior, L929 cells were seeded into 96-well plates  in growth medium at 30,000 cells/cmbrushing ,15. AfteHalf lethal concentration values (LC50) of the toothpastes are reported (mean and standard deviation) from two independent experiments, each performed in triplicate. Differences in LC50 between cells treated with the five toothpastes were tested using a non-parametric Kruskal–Wallis test followed by a post-hoc Mann–Whitney U-test with Bonferroni correction for multiple comparisons . Alpha of 5% was considered significant. The results on the cell proliferation behavior are described by the mean and standard deviation.AirFlow was the only toothpaste showing a slightly basic pH value (7.75). The four commercially available toothpastes all showed a slightly acidic pH, with OB having the lowest value at 5.57. The SLS-free Ze also showed a low value of 6.08. The remaining two toothpastes, OM and Se, had nearly similar pH values of 6.75 and 6.69, respectively. Regarding the concentration of free fluoride, the SLS-free, commercially available toothpaste Ze had the highest value of 1575 ppm. The experimental AF toothpaste had 1400 ppm and the three remaining commercially available toothpastes had 1290 ppm (OM), 1106 ppm (Se), and 1050 ppm (OB) .p < 0.01). Only in comparison to the second SLS-free toothpaste (Zendium), AF showed no statistically significant difference in the LC50 values. Comparing the commercially available toothpastes only, Ze showed significantly higher LC50 values than OB and OM (both p = 0.03). Compared to Se, Ze also showed higher LC50 values, but without statistical significance after Bonferroni correction. The LC50 values of the toothpastes on the primary cells of the oral cavity, gingival fibroblasts (GF), and periodontal ligament cells (PDL) confirm the results obtained from the L929 cell tests.The half lethal concentration (LC50) of toothpaste-conditioned medium (TCM) of the toothpastes  on L929 cells, gingival fibroblasts (GF), and periodontal ligament cells (PDL) are shown in w/v% AirFlow TCM, all commercially available toothpastes remained below a proliferation rate of 26%. When incubating the cells with 3% TCM, AF even resulted in unimpaired proliferation behavior when compared to the untreated control, while the toothpastes OM, OB, Se, and also Ze were still strongly impaired in their proliferation behavior  as a surfactant, the commercially available toothpaste Zendium and the experimental toothpaste AirFlow used alternative surfactants.The chemical parameters investigated included the measurement of pH and free fluoride concentration, while the biological parameters included the calculation of the half lethal concentration of the toothpastes in cell culturing tests and the investigation of the proliferation behavior of the cells. Remarkably, the newly developed toothpaste (AF) and the second SLS-free toothpaste (Ze) showed the best results in terms of compatibility for the cells investigated. Regarding the free fluoride content, AirFlow and Zendium, with a detected fluoride concentration of 1400 ppm and 1575 ppm, respectively, were in the upper range of the WHO recommendation or even above. OM, with a free fluoride content of 1050 ppm, was at the lower end of the recommendation, and toothpastes OB and Se, with free fluoride concentrations of 1290 ppm and 1100 ppm, were in the middle range. However, the similarities between the two SLS-free toothpastes, AF and Ze, did not extend to the measured pH value. While AF exhibited the highest basic pH value of 7.75, Ze showed an acidic pH value of 6.08. While OM had the lowest pH value of 5.57, OB and Se showed similar values of 6.75 and 6.69 also for this parameter.Regarding fluoride concentration, it must be noted that all toothpastes tested had proven fluoride concentrations above 1000 ppm. Although the caries-protective effect of the investigated toothpastes was not investigated in the present study, the protection against caries by the use of fluoride-containing toothpastes above a concentration of 1000 ppm has been confirmed by numerous studies and consequently by a systematic review by the Cochrane Collaboration . AdditioFinally, the question arises as to the significance of the individual parameters for the clinic. Looking at the chemical parameters of the study, it can be seen that all except one toothpastes complied with the WHO recommendations regarding the fluoride concentration and the International Organization for Standardization (ISO) recommendations regarding pH. However, it should be noted that a new measurement method was described in the literature explaining higher pH values in the original toothpaste than in toothpaste slurries . WhetherIn addition to the actual requirements that toothpaste must fulfill, such as cleaning the teeth, releasing fluoride, and much more, toothpaste must also be well tolerated by the tissue surrounding the teeth. Regarding the biological parameters of the toothpastes studied, the calculation of the half lethal concentration  was performed on different kinds of cells. AirFlow showed significantly higher LC50 values, which indicated its lower toxicity compared to the three commercially available toothpastes . This confirms the good tolerance of the toothpaste at the cell biological level. The fourth commercially available toothpaste (Ze) also showed high LC50 values. One explanation for the better cell survival after stimulation with AF and Ze could be the presence of alternative surfactants than sodium lauryl sulfate, which has already been shown by other studies ,12.w/v% toothpaste slurry. This additional positive cell-protective function might be provided in AirFlow by the addition of erythritol. Erythritol was previously shown to protect endothelial cells under hyperglycemic conditions [Regarding the proliferation behavior of the cells after incubation with the respective toothpastes, the results of the viability tests are confirmed insofar as the cells after incubation with AF showed higher proliferation behavior compared to the cells incubated with toothpastes containing SLS. Interestingly, cells after incubation with AF showed higher proliferation rates compared to cells treated with the SLS-free toothpaste, Ze. The toothpaste AF resulted in a cell proliferation rate of 87% while Ze, known for its good biocompatibility, resulted in 26% after incubation with 6 nditions . In addinditions . In the nditions . Despitenditions . Until nnditions . Anothernditions . Since tIn conclusion, the newly developed erythritol- and xylitol- containing toothpaste AirFlow shows excellent biocompatibility in cell tests and has high availability of free fluoride. Further studies on other important points for a toothpaste in daily use, such as the effectiveness in removing plaque, the effect on bacteria, as well as the performance in a caries model, are of course necessary and in planning.


Background and Purpose: To assess the safety and effectiveness of oral methylprednisolone (oMP) in comparison with intramuscular adrenocorticotropic hormone (imACTH) and oral prednisolone (oP) therapies in children with infantile spasms (IS).Methods: In this prospective, open-label, non-blinded, uncontrolled observational study, children (aged 2–24 months) with newly diagnosed IS presenting with hypsarrhythmia or its variants on electroencephalogram (EEG) were included. It was followed by imACTH, oP, or oMP (32–48 mg/day for 2 weeks followed by tapering) treatments. Electroclinical remission/spasm control, relapse, and adverse effects were evaluated in the short-term (days 14 and 42) and intermediary-term  intervals.Results: A total of 320 pediatric patients were enrolled: 108, 107, and 105 in the imACTH, oMP, and oP groups, respectively. The proportion of children achieving electroclinical remission on days 14 and 42 was similar among the three groups . The time to response was significantly faster in the oMP group . Spasm control at 3, 6, and 12 months was also similar in the three groups . The relapse rate in the imACTH group (24.10%) was lower than oMP (30.77%) and oP groups (33.33%), and the time taken for relapse in the imACTH group  was longer than oMP  and oP groups , but the differences were not statistically significant . The occurrence of adverse effects was similar among the three groups.Conclusions: The short and intermediary-term efficacy and recurrence rates of oMP are not inferior to those of imACTH and oP for the treatment of IS. Significantly, the time to achieve electroclinical remission with oMP was quicker than that with imACTH and oP. Considering its convenience, affordability, and the absence of irreversible side effects, oMP can serve as a form of first-line treatment for newly diagnosed IS. Infantile spasm (IS), a subtype of epileptic spasms, is an epileptic encephalopathy and is frequently associated with hypsarrhythmic interictal electroencephalograms (EEG) and psychomotor retardation . The incThe United Kingdom Infantile Spasms Study suggested that the ideal treatment agent for IS should achieve rapid control of spasms, normalize the EEG changes early, maintain a spasm-free state, and thus contribute to a better intermediary-term developmental outcome . CurrentAccording to the reports of the Sri Lanka Infantile Spasms Study (SLISS), the two most commonly used forms of hormonal therapies, the intramuscular adrenocorticotropic hormone  and the oral prednisolone , exhibit no difference in the short- (28 days) and intermediary-term  control of newly diagnosed IS and can be considered as equal options for first-line treatment , 10. HowThe reported efficacy of ivMP (30 mg/kg/day) in children with IS is ~60–83%, with freedom from spasms being achieved very early (2–3 days following treatment) among responders , 13. RecThis study was designed as a prospective, open-label, non-blinded, uncontrolled observational study to evaluate the efficacy and tolerability of oMP in children with newly diagnosed IS. From January 2016 to October 2020, children aged 2–24 months with newly diagnosed epileptic spasms in clusters with electroencephalographic evidence of hypsarrhythmia or its variants were enrolled and assessed for eligibility in the Department of Neurology of Wuhan Children's Hospital. Children with a diagnosis of IS based on the definition proposed by Lux et al.  and thosBefore treatment allocation, the baseline characteristics of all eligible participants were assessed using detailed clinical histories and examinations. Participants' ages at onset, perinatal details, family histories, and developmental statuses were recorded. The results of investigations such as routine blood tests, liver and kidney function, blood electrolytes, arterial blood gas, blood lactate, blood ammonia, urine ketones, blood tandem mass spectrometry, neuroimaging , EEG, metabolic testing, and Gesell development schedules test were documented. The etiology of IS was classified according to six etiologic categories defined by the International League Against Epilepsy Task Force, including structural, genetic, infectious, metabolic, immune, and unknown .The parents or guardians of all eligible participants were provided with detailed information about the standard treatment options available for IS, including the total cost, efficacy, and potential adverse effects of imACTH, oP, and oMP therapies. They were asked to make an informed choice based on the feasibility and expense of the three treatments. A chest X-ray and tuberculin test were performed to screen for tuberculosis before starting hormonal therapy. The baseline weight and blood pressure were measured, and pyridoxine was administered to exclude pyridoxine-dependent seizures.The imACTH group received ACTH  intramuscularly (25 IU/day) for 2 weeks. If spasms continued until day 7 or reappeared from days 8 to 14, the dose was increased to 40 IU/day. After 2 weeks of treatment, all children received oral prednisolone  at an initial 2 mg/kg dose, which was tapered off and stopped over 4 weeks. The oP group received oral prednisolone at a dose of 10 mg four times/day for 2 weeks. If spasms continued until day 7 or reappeared from days 8 to 14, the dose was increased to 20 mg three times/day. Prednisolone was tapered and stopped over 4 weeks: reductions were made once a week, with 30 mg daily, then 20 mg, then 10 mg, and then 5 mg; if on the higher dose of treatment, the doses were 40 mg daily, then 20 mg, then 10 mg, and then 5 mg.The oMP group received oral methylprednisolone  at a dose of 8 mg four times/day for 2 weeks. If spasms continued until day 7 or reappeared from day 8 to day 14, the dose was increased to 16 mg three times/day. After 2 weeks of treatment, methylprednisolone was tapered and stopped over 4 weeks: reductions were made once a week, with 24 mg daily, then 16 mg, then 8 mg, and then 4 mg; if on the higher dose of treatment, the doses were 32 mg daily, then 16 mg, then 8 mg, and then 4 mg.Currently, there are no guidelines or expert consensus for the subsequent treatment of IS following hormone therapy. In this work, all patients received oral topiramate  after the end of 6 weeks of hormone therapy.After the initial treatment, patients continued to be followed up at different time points. The initial follow-up included reviews at 14 and 42 days posttreatment, followed by subsequent reviews at 3, 6, and 12 months posttreatment. The first two time points were considered markers of a short-term response, and the three later time points were considered markers of an intermediary-term response. Parents or guardians were trained to document spasms and maintain seizure diaries. At all follow-up visits, the children were evaluated by the same pediatric neurologist. Those who relapsed were treated again with hormonal therapies or other oral anticonvulsants at their parents' discretion. Drug compliance was monitored by asking the parents or guardians directly. The numbers of patients who did not present for follow-up and those who died during follow-up were noted separately for the different time points.The primary outcomes measured were the short- and intermediary-term responses. The evaluated outcome indicators for short-term response were the number of children with electroclinical remission  at the time of review on days 14 and 42.The evaluated outcome indicators for intermediary-term response were the number of children with spasm control (defined as the absence of witnessed spasms for ≥28 consecutive days from the time of the last witnessed spasm) at 3, 6, and 12 months after initial therapy.The secondary outcomes measured were the time to response (defined as the first day after initiation of treatment on which spasms were not seen) and relapse , and the time taken for relapse .Adverse effects (defined as any untoward or unintended responses) were thought to be related to the treatments. The anticipated potential adverse reactions included cushingoid features, weight gain, increased appetite, frequent crying spells, irritability, infections, drowsiness, gastrointestinal upset , and electrolyte imbalances. Other parental concerns regarding side effects were also noted.Categorical data are summarized in proportions, and quantitative data are presented as medians and quartiles. The significance of the differences in electroclinical remission and relapses between the three treatment groups were assessed at 14 and 42 days and 3-, 6-, and 12-month intervals using the Chi-square test. The effect size was calculated using the risk ratio  and absolute risk reduction  with 95% confidence intervals (CIs). The number needed to treat (NNT) was calculated based on the number of patients who needed methylprednisolone to achieve spasm remission, compared with a patient who did not. The Kaplan–Meier survival method was used to plot the relapse and response rates under different groups. The significance level was set at 0.05, and all tests were two-tailed. Statistical analyses were performed using SPSS version 22.0.This study was reviewed and approved by the Medical Ethics Committee of Wuhan Children's Hospital, Tongji Medical College, Huazhong University of Science & Technology (2015019). The parents of the patients signed written informed consent, agreed with their children's participation in this study, and allowed the use of the relevant data and information for scientific research.A total of 335 children with clinically confirmed epileptic spasms were evaluated. Eventually, only 320 children were included in the study to compose the three treatment groups. A total of 108, 107, and 105 children were allocated to the imACTH, oMP, and oP groups, respectively. The baseline characteristics of the three groups showed no significant differences .p = 0.869), 3 (p = 0.801), 6 (p = 0.685), and 12 months (p = 0.791). Four deaths occurred during the 12-month follow-up, all of which occurred after completing the 6-week initial treatment and were related to other systemic diseases.Details of enrollment, treatment allocation, and follow-up  are outlined in p = 0.362). Electroclinical remission by the end of day 42 was higher in the oMP group  than in the imACTH  and oP groups , although the difference was not statistically significant (p = 0.470).The time course of treatment and outcome evaluation is presented in P = 0.025). The proportion of infants with an electroclinical remission and the corresponding required days within 42 consecutive days post initial treatment is graphically presented in The total number of days required for electroclinical remission was significantly lower in children treated with oMP  compared with those treated with imACTH  and oP  (P = 0.775). In addition, the proportions of children in whom spasm control was achieved also showed no significant difference at 6 (p = 0.667) and 12 months (p = 0.779) in the three groups , oMP , and oP  groups showed no significant difference (e groups .p = 0.539). In addition, no significant difference was noted in the time taken for relapse within each treatment group (p = 0.530), although children within the oMP group  tended to relapse earlier than those in the imACTH  and oP groups  during the 12-month follow-up period . AlthougAdverse effects observed in the three groups included cushingoid features, weight gain, increased appetite, frequent crying spells, irritability, drowsiness, increased susceptibility to infection, gastrointestinal upset , and electrolyte imbalances . The mosThe most accepted standard first-line treatment for IS is hormonal therapy, which mainly refers to imACTH and oP. ACTH is preferred but is expensive and difficult to obtain in some countries. Several trials and meta-analyses have confirmed that high-dose oP (40–60 mg/day) is not inferior to ACTH and is a more cost-effective treatment for IS , 21, 22.2/day) was associated with a greater early response rate than oP by analyzing the multicenter database of the National Infantile Spasms Consortium, whereas Wanigasinghe et al. found that the time required for spasm cessation was quicker in children treated with oP compared with imACTH at a dose of 40–60 IU every other day (3.85 vs. 8.65 days) in the SLISS trial. In this study, the total number of days required for spasm control was ~8 days in children treated with oP and imACTH (25–40 IU/d). In addition, oP and imACTH showed no significant difference in the time until a response in 3/5 randomized controlled trials, and the mean time to spasm cessation ranged from 3 to 8 days in a metaanalysis conducted by Kuo et al.  of the SLISS trial conducted by Wanigasinghe et al. . Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin.Z-SL, CY, J-SH, and G-FW: study conception and design. H-MZ, M-QL, SH, and X-LD: data acquisition. C-HY, CY, and H-MZ: analysis and data interpretation. H-MZ and C-HY: drafting of the manuscript. All authors contributed to the article and approved the submitted version.This work was supported by the Wuhan Municipal Health Commission of Clinical Research Projects (NO. WX17B11).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.


Toxocara canis and Toxocara cati, the roundworms of dogs and cats. Infection may result in the clinical picture of toxocarosis, one of the most common zoonotic helminthoses worldwide. To estimate the Toxocara seroprevalence in four different villages of the Wiwa community, serum samples from 483 inhabitants were analyzed for anti-Toxocara-antibodies. Overall, 79.3% (383/483) of analyzed samples were seropositive. Statistically significant differences were observed between the four villages, as well as age groups (adults > adolescents > children), while sex had no effect. The high seropositivity rate demonstrates the risk of zoonotic roundworm infections and potential clinical disease in vulnerable indigenous inhabitants.The life of the indigenous Wiwa tribe in northeast Colombia is characterized by lacking access to clean drinking water and sanitary installations. Furthermore, free-roaming domestic animals and use of yucca and/or manioc as a primary food source favor the transmission of soil-transmitted helminths, e.g.,  Toxocara canis and Toxocara cati, the zoonotic roundworms of dogs and cats. Furthermore, a lack of access to clean water and sanitary installations affects environmental hygiene as well as personal hygienic standards, providing ideal transmission possibilities for STHs [Ancylostoma spp., Necator americanus, Strongyloides stercoralis, Ascaris lumbricoides, Trichuris trichiura, and Hymenolepis nana were detected in members of the Wiwa community [The Sierra Nevada de Santa Marta in the northeast of Colombia presents a great variety of climates and tropical ecosystems within a relatively small area. The ethnohistorical habitation of the region dates back at least to the 1st century AD with the pre-Columbian culture of the Tairona, of which the Kankuamo, Kogi, and Arhuacos, as well as the Wiwa, are the believed direct descendants . To avoifor STHs ,4. Conseommunity .Toxocara spp. [Toxocara antibodies in humans is reported for some tropical and subtropical regions or countries [Toxocara spp. third-stage larvae (L3) in contaminated food, water, and soil, or via consumption of L3 in undercooked meat of paratenic hosts such as chicken [Additionally, environmental conditions, such as annual mean temperatures of 25–30 °C, high humidity, and soil moisture, are favourable for the development and tenacity of a broad range of (zoonotic) helminth eggs, such as ara spp. ,7,8. Conountries . Toxocarountries , and hum chicken . By excr chicken .Toxocara larvae into the brain [Toxocara spp. As clinically apparent toxocarosis and its consequences affect the life of the individuals, the presented study aimed to estimate the seroprevalence in the indigenous tribe of the Wiwa, and to analyse potentially related risk factors. The results are necessary to gain knowledge about the risk of exposure, and to raise awareness for this neglected disease in indigenous communities.Infections may remain asymptomatic; however, the migration and persistence of larvae in the tissues can also result in clinical disease, ranging from mild to severe, and rarely fatal organ injuries . Overallhe brain ,16. OLM he brain ,18. The he brain . Thus faSample material was obtained in accordance with the international guidelines of the Declaration of Helsinki, approved by the local ethics committee of Valledupar, Cesar, Colombia (Acta no. 0022013). Written informed consent was obtained from each participant or the parent/legal guardian of a child prior to participation.n = 173), Ashintukwa (n = 106), Cherua (n = 94), and Siminke (n = 110) were collected. Participants were categorized by their age  and sex . Furthermore, the villages were categorized regarding altitude, being categorized as below 1000 m above sea level (MASL)  and above 1000 MASL . Collected blood samples were cooled in a specific cooling box at 4 °C and transported immediately to the laboratory for serum extraction. Sera were stored thereafter at −20 °C and transferred to Germany with a permanent cooling chain. After arrival, sera were stored at −20 °C until further analysis.In July and November 2014, blood samples of 483 indigenous volunteers from the Wiwa villages of Tezhumake , based on Toxocara excretory-secretory products (TES) as diagnostic antigen, was conducted in accordance with the manufacturer’s instructions. The described diagnostic specificity and sensitivity is given as 98.63% (95%-CI (96.52–99.62%)) and 96.92% (95%-CI (89.32–99.63%)). All runs included a positive and negative control, as well as a calibrator for semi-quantitative analysis. Briefly, all samples were diluted 1:101 with supplied IgG sample diluent, and a volume of 100 µl standards/controls as well as diluted sera were incubated as duplicates for 1 h at 37 °C. After incubation and washing steps consisting of three times 300 µl washing buffer, 100 µl horseradish peroxidase-labelled conjugate was added and again incubated for 30 min at 37 °C. After washing as described, colorimetric detection was initiated by applying 100 µl tetramethylbenzidine substrate solution for 30 min at room temperature and stopped with 100 µl stop solution. The optical density (OD) was measured using a Biowave 340 photometer  at a wavelength of 450 nm as well as a reference wavelength of 620 nm to exclude background signals. Result in units (U) were calculated, and samples were tested negative if U was <9, equivocal if U was ≥9 to <11, or positive if U was ≥11.To determine anti-Toxocara seroprevalence differences in the Wiwa, a general linear model (GLM) with binomial error structure and logit link function was conducted. Village, age group, and sex were included as fixed factors, and multiple comparisons between these factors were carried out using Tukey HSD contrasts with single-step p-value adjustment . Final Toxocara antibodies were detected in 79.3% of the 483 analyzed samples, and in 3.3%, the results were equivocal. In case of equivocal results, the manufacturer recommends repetition with freshly obtained serum after 2 to 4 weeks. As repetition of equivocal results was not possible, these samples were judged as negative to fit in the GLM. Anti-The highest seroprevalence was detected in the village of Siminke (90.0%), the lowest in Tezhumake 70.5%, . SubgrouToxocara seroprevalences differed significantly between the communities. Overall, the seroprevalence in villages below 1000 MASL (Tezhumake and Ashintukwa) was statistically significantly lower compared to villages above 1000 MASL. Statistically significant differences were also determined between the age classes: the seroprevalence in adults was significantly higher than in adolescents and children, while sex had no effect on seropositivity. Detailed results are presented in Toxocara spp., their high reproduction rate, and the high tenacity of the eggs result globally in soil contamination of human environments [Toxocara eggs, soil contamination rates are elevated at lower latitudes (0–20°) compared to higher ones (41–60°) [Toxocara prevalence with, estimated on the population size, high burdens of human toxocarosis [Toxocara antibodies. Notably, the seroprevalence of 79.3% in the northeast Colombian indigenous Wiwa tribe investigated in the present study is remarkably higher than previously described for Colombia.The worldwide distribution of ronments ,23, makironments . As trop(41–60°) . Consequocarosis . Colombiocarosis . Howeverocarosis , as 42.1Toxocara infections, such as living in rural areas, having contact with dogs, cats, or soil, and the drinking of untreated water [Toxocara egg contamination of different vegetables such as lettuce, carrots, potatoes, zucchini, spinach, cucumber, or cress have been described [Toxocara infections.Besides tropical climate conditions, various further potential risk factors have been associated with human ed water . As the ed water . Their led water ,4. Furthed water . Such hiescribed ,28,29. TToxocara eggs, which may survive for up to four years in warm and humid environments [The seroprevalence in the Wiwa population varied between the different villages. The seroprevalences of 88% and 90% in inhabitants of Cherua and Siminke were significantly higher compared to those of Tezhumake and Ashintukwa (70% and 75%). Although all communities have difficult living conditions, regional climatic conditions differ between the four villages. The daily average temperature in all four communities is above 18 °C, but Tezhumake and Ashintukwa are located at 700 and 750 MASL, and the climate at this altitude is characterized by a prolonged dry season of up to six months a year. In contrast, Cherua and Siminke are located at 1300 and 1100 MASL, where higher precipitation and higher average humidity and a subtropical or premontane humid forest result in an increased soil moisture. This favors the development and survival of embryonated ronments ,32. ThusToxocara seropositivity in the Wiwas was associated with increasing age. However, age as a risk factor for human seropositivity remains debatable. Although children are regarded as being at high infection risk due to a less developed hygienic awareness compared to adults [Toxocara antibodies in elderly people may result from a cumulative effect of Toxocara exposure during their lifespan. The constant rechallenge of the immune response due to less developed hygienic standards, leading to frequent and/or high Toxocara exposure of all inhabitants regardless of age, may result in the accumulation of detectable antibodies. On average, IgG antibodies against Toxocara can be detected for 2.7 years with TES-ELISA [Toxocara larvae has been associated with mental confusion and cognitive impairment, as well as deficits in the development of speech and poor reading achievements, a higher distractibility, and lower intelligence in kindergarten children [Furthermore, o adults ,33, elevES-ELISA  as used ES-ELISA ,35,36. Nchildren ,18,37,38Toxocara exposure [Toxocara eggs due to increased agricultural activities, and thus increased contact with Toxocara egg-contaminated soil and stray animals [Toxocara eggs.Similar to age as an influencing factor, the impact of sex is controversially discussed. The majority of studies have identified males as being at higher risk for exposure ,39,40, bexposure . For mal animals ,43. In tToxocara eggs in community environments would be an important measure. This comprises the avoidance of pet defecation in the villages and farmland and/or disposal of feces in these places [Overall, the high seroprevalence of 79.3% in indigenous Wiwa indicates a high health risk of exposed people . As infee places ,47. HoweToxocara infections are potentiated by nutrition habits and free-roaming dogs and cats in the Wiwa communities, resulting in a high seroprevalence of about 80% in the tested inhabitants. Deviating climatic conditions in the included villages are most likely responsible for the higher seroprevalences in communities above 1000 MASL with a subtropical or premontane humid forest compared to the lower altitudes with up to six months of dry season. Nevertheless, frequent Toxocara exposure is a common feature in all Wiwa communities, reflected by a cumulative effect with highest seroprevalences in adult inhabitants, and lacking sex differences. As toxocarosis can result in detrimental effects and long-term consequences, raising awareness thereof, and intervention measures, such as improvement in hygienic standards and reduction in soil contamination, are urgently needed in indigenous communities such as the Wiwa tribe.The secluded life of the indigenous tribe of the Wiwas in north Colombia is characterized by, for example, a lack of access to clean drinking water and adequate hygiene standards. These well-known risk factors for zoonotic


Most solid tumours are hypoxic. Tumour cell proliferation and metabolism accelerate oxygen consumption. The low oxygen supply due to vascular abnormalisation and the high oxygen demand of tumour cells give rise to an imbalance, resulting in tumour hypoxia. Hypoxia alters cellular behaviour and is associated with extracellular matrix remodelling, enhanced tumour migration, and metastatic behaviour. In light of the foregoing, more research on the progressive and prognostic impacts of hypoxia on gliomas are crucial. In this study, we analysed the expression levels of 75 hypoxia-related genes in gliomas and found that a total of 26 genes were differentially expressed in The Cancer Genome Atlas (TCGA) database samples. We also constructed protein–protein interaction networks using the STRING database and Cytoscape. We obtained a total of 10 Hub genes using the MCC algorithm screening in the cytoHubba plugin. A prognostic risk model with seven gene signatures  was constructed based on the 10 Hub genes using LASSO–Cox regression analysis. The model was verified to be highly accurate using subject work characteristic curves. The seven-gene signatures were then analysed by univariate and multivariate Cox. Notably, PSMB10, PSMD12, UBB, PSMA5, and PSMB6 were found to be independent prognostic predictive markers for glioma. In addition, PSMB6, PSMA5, UBB, and PSMD12 were lowly expressed, while PSMB10 was highly expressed, in the TCGA and GTEx integrated glioma samples and normal samples, which were verified through protein expression levels in the Human Protein Atlas database. This study found the prognostic predictive values of the hypoxia-related genes PSMB10, PSMD12, UBB, PSMA5, and PSMB6 for glioma and provided ideas and entry points for the progress of hypoxia-related glioma. There are more than 100 histological subtypes of primary brain and central nervous system (CNS) tumours . CurrentTumour hypoxia is a condition in which tumour cells are deprived of oxygen . During Cancer cells have multiple mechanisms for evading radiotherapy-induced cell death. Among them, the development of tumour hypoxia and its associated metabolic pathways is one of the most important contributors to clinical radioresistance . This isIn this study, we analysed the expression and correlation of 75 hypoxia-related genes in gliomas and thereafter constructed a highly accurate prognostic risk prediction model consisting of seven gene signatures. PSMB10, PSMD12, UBB, PSMA5, and PSMB6 were found to be independent predictors of glioma prognosis.https://portal.gdc.cancer.gov/) website, while the mRNA expression data of 2,642 cases of normal tissues were downloaded from the Genotype-Tissue Expression  website. The 75 hypoxia-associated genes were cited in Wei et al. (The data of 663 glioma (GBM + LGG) samples, and mRNA expression data, were downloaded from The Cancer Genome Atlas  was used to analyse the differences in the 75 hypoxia-associated gene expressions. Correlations between quantitative variables without a normal distribution were described using Spearman’s correlation analysis. p < 0.05 was considered statistically significant.Consistency analysis was performed using the R package ConsensusClusterPlus (v1.54.0) with a maximum number of clusters of six and 100 replicates to extract 80% of the total sample, clustering = “hc”, innerLinkage = ‘ward.D2’. The clustering heatmaps were all analysed using the R software package pheatmap (v1.0.12). The gene expression heatmaps were retained for genes with variances above 0.1.https://string-db.org/) was used to identify known and predicted PPIs. Seventy-five hypoxia-associated genes were analysed, and PPI networks were constructed using STRING. The top 10 Hub genes in the PPI networks were further screened using cytoHubba in Cytoscape (v3.8.2) software.The STRING database  with 95% confidence intervals (CI) in the Kaplan–Meier curves were derived through log-rank test and univariate Cox proportional hazards regression.The LASSO regression algorithm was used for feature selection, and 10-fold cross-validation was used to determine the parameters needed to obtain a suitable model. The genes obtained from LASSO regression were then subjected to multifactor Cox regression analysis, and the multifactor regression coefficient of each gene was calculated to construct the risk score equation. The patients were divided into high- and low-risk groups according to the median risk score values. The Kaplan–Meier survival curve analysis was used to compare the overall survival times of the two groups, and the predictive value of the genetic markers was evaluated through time-related receiver operating characteristic (ROC).Cox regression analysis was performed using the Survival package, and forest plots were plotted using the forestplot package to obtain the p-value, HR, and 95% CI for each variable. Based on the results of the multivariate Cox proportional risk analysis, column line plots were constructed using the RMS package to predict the 1-, 3-, and 5-year survival rates.Immunohistochemical staining maps of the gene expression in both glioma tissues and normal tissues were downloaded from the Human Protein Atlas (HPA) database.Samples were divided into two groups of high and low expression according to the median value of gene expression, and the enrichment of Kyoto Encyclopedia of Genes and Genomes (KEGG) and HALLMARK pathways in the high and low expression groups were analysed using gene set enrichment analysis (GSEA).The analysis results of the expression levels of 75 hypoxia-related genes in 663 glioma samples and five paraneoplastic tissue samples from the TCGA database showed that EGLN2, PSMD1, HIF1AN, PSMD10, PSMB10, ELOB, PSME2, PSMF1, AJUBA, PSMB4, LIMD1, PSMC6, PSMB1, PSMB8, ARNT, GUL2, PSMA3, SEM1, EPAS1, PSMA2, EPO, PSME3, PSMB9, HIF1A, UBA52, and RPS27A were significantly differentially expressed in cancer and paraneoplastic tissues  of the 20S proteasome core complex. This proteasome is a large multisubunit complex that is involved in protein degradation of PSMA5 , 31. Preia model . PSMB6 iia model . In addiia model –35. PSMBia model . In the ia model , 38. Disia model . In addiia model  construcia model . The comia model , 43. Simin vivo or in vitro.Combining the above findings, we can identify the important roles of PSMB10, PSMD12, UBB, PSMA5, and PSMB6 in gliomas, thus providing new targets and ideas for tumour-targeted therapy. The present study has some limitations. Given that there is a dearth of research analysing the genes in tumours, the discovery of their mechanisms of action still needs improvement. Therefore, more biological experiments are needed to prove whether the conclusions reached can be verified In summary, we constructed a prognostic model for glioma based on seven hypoxia-related genes and further identified five independent predictors of prognosis in glioma patients, thereby providing potential new targets for glioma gene-targeting therapy.The original contributions presented in the study are included in the article/All authors listed have made substantial, direct, and intellectual contribution to the work and approved it for publication.This study was supported by Ningbo public welfare science and technology program (No. 20181JCGY020333).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. 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Objective: The occurrence, development, and prognosis of serious adverse events (SAEs) associated with anticancer drugs in clinical trials have important guiding significance for real-world clinical applications. However, to date, there have been no studies investigating SAEs reporting in randomized clinical trials of colorectal cancer treatments. This article systematically reviewed the SAEs reporting of phase III randomized clinical trials of colorectal cancer treatments and analyzed the influencing factors.Methods: We reviewed all articles about phase III randomized clinical trials of colorectal cancer treatments published in the PubMed, Embase, Medline, and New England Journal of Medicine databases from January 1, 1993, to December 31, 2018, and searched the registration information of clinical trials via the internet sites such as “clinicaltrials.gov”. We analyzed the correlation between the reported proportion (RP) of SAEs in the literature and nine elements, including the clinical trial sponsor and the publication time. Chi-square tests and binary logistic regression were used to identify the factors associated with improved SAEs reports. This study was registered on PROSPERO.Results: Of 1560 articles identified, 160 were eligible, with an RP of SAEs of 25.5% (41/160). In forty-one publications reporting SAEs, only 14.6% (6/41) described the pattern of SAEs in detail. In clinical trials sponsored by pharmaceutical companies, the RP of SAEs was significantly higher than that in those sponsored by investigators . From 1993 to 2018, the RP of SAEs gradually increased (none (0/6) before 2000, 17.1% (12/70) from 2000 to 2009, and 34.5% (29/84) after 2009). The average RP of SAEs published in the New England Journal of Medicine (N Engl J Med), the Lancet, the Journal of the American Medical Association (JAMA), the Lancet Oncology (Lancet Oncol), and the Journal of Clinical Oncology (J Clin Oncol) was significantly higher than that published in other journals . In the clinical trials referenced by clinical guidelines, the RP of SAEs was higher than that in non-referenced clinical trials . Binary logistic regression analysis showed that pharmaceutical company sponsorship, new drug research, and sample size greater than 1000 were positive influencing factors for SAEs reporting.Conclusion: Although the RP of SAEs increased over time, SAEs reporting in clinical trials needs to be further improved. The performance, outcomes and prognosis of SAEs should be reported in detail to guide clinical practice in the real world. Colorectal cancer is the third most commonly diagnosed malignancy worldwide . ChemothAEs reporting is relatively higher in cancer clinical trial publications, but the reporting quality is low. A review showed that 96% of cancer clinical studies reported AEs, but oncology-specific reporting standards were lacking . AnotherWe systematically reviewed SAEs reporting from publications of colorectal cancer clinical trials, to further draw researchers’ attention to SAEs reporting. The SAEs reporting was influenced by many social factors, such as regional policy, preciseness and awareness of investigators, purpose of sponsor, so this article analyzed the possible influencing factors of SAEs reporting. Because the results of phase III randomized clinical trials were the most instructive in the real world, herein we just reviewed phase III randomized clinical trials.This study included randomized phase III colorectal cancer clinical trials whose intervention measures contained anticancer pharmaceuticals and whose results were published in PubMed, Embase, Medline, and the New England Journal of Medicine from January 1, 1993, to December 31, 2018. We analyzed several possible factors that may affect SAEs reporting in the literature. These factors included the region where the clinical trial was conducted, the sponsor of the clinical trial, whether the trial researched new drugs, the publication date which may reflect the change of policy and awareness of investigators, factors related with the rigorous of the clinical trials such as sample size, the type of journal and whether the clinical guidelines referenced the results of the study, and factors owned by clinical trials themselves, such as treatment line, therapeutic schedule.via the following internet sites: http://www.clinicaltrials.gov, http://www.isrctn.com/search, http://www.anzctr.org.au, https://www.umin.ac.jp/, http://apps.who.int/en/. The inclusion criteria were as follows: 1) phase III randomized colorectal cancer clinical trials, 2) intervention measure contained chemotherapy and/or target therapy, 3) the articles showed the efficacy and/or safety of the clinical trial, 4) published in English. The exclusion criteria were as follows: 1) the same research published repeatedly, 2) reviews, meta-analysis, molecular analysis and cost analysis, 3) subgroup analysis of the research already included, 4) intervention measure contained immune therapy (because the AEs spectrum of chemotherapy and immunotherapy is different), 5) clinical trials aimed to observe the efficacy or AEs of accompanying regimens along with anticancer therapy. The primary objective was the reported proportion (RP) of SAEs. The secondary objectives were the performance, outcomes and prognosis of SAEs.A review of citations from PubMed, Embase, Medline, and New England Journal of Medicine for studies published between January 1, 1993 and December 31, 2018, was performed to identify eligible colorectal cancer clinical trial publications for the analysis. The search terms were as follows: “colorectal cancer” [All fields] or “colon cancer” [All fields] or “rectal cancer” [All fields], and “phase 3” [All fields] or “phase III” [All fields]. We used the filters as follows: “subjects = cancer,” “article type = clinical trial,” “language = English,” “species = humans,” and “publications dates = 1/1/1993-12/31/2018.” Endnote X4  was used to manage the publications. We searched the registration information of clinical trials According to the National Cancer Institute Common Toxicity Criteria (NCI-CTC) version 5.0 , an AE iSAEs were diagnosed according to NCI-CTC version 5.0  as folloAn SAE is any untoward medical occurrence that, at any dose: 1) results in death, 2) is life-threatening, 3) an event is considered life-threatening if it is suspected that the individual is at substantial risk of dying at the time of the AEs, 4) requires inpatient hospitalization or prolongation of existing hospitalization , 5) results in persistent or significant disability/incapacity , 6) is a congenital anomaly/congenital disability, 7) is medically significant .Herein we mainly discussed the SAEs reporting. If the publication pointed out the occurrence of SAEs, even the incidence was zero, it was judged to have reported SAEs. SAEs consisted of many events not only death, so if the publication just only reported death and didn’t mention “SAEs,” it wasn’t judged to have reported SAEs in this review. And reporting Grade 3/4 AEs were not identified as having reported SAEs.The data were collected independently by two investigators (Yanhong Yao and Zhentao Liu) who screened eligible publications and searched the registry of clinical trials. The collected data included performance, outcomes and prognosis of SAEs, the region where the clinical trial was conducted, the sponsor of the clinical trial, whether the trial researched new drugs, the sample size, the publication date, the type of journal, whether the clinical guidelines  referenced the study results, the treatment lines, and treatment schedules. Professor Baoshan Cao checked the data if inconsistencies existed between the results collected by the two investigators.p values were less than 0.05. Frequencies and percentages were calculated for counting data. The chi-square test was used to assess the association between RPs of SAEs and collected items, and Fisher’s exact test was used if the theoretical number was less than 5 or the sample size was less than 40. A binary logistic regression model was used to identify items associated with SAEs reporting. The dependent variable was whether reported SAEs, and the independent variables were the positive influencing factors for SAEs reporting based chi-square test. The method of the independent variables entering the regression equation was “Backwald”.We used SPSS version 19.0  to analyze the data, and differences were considered statistically significant when the two-sided From the 1560 publications initially collected by the two investigators, a total of 160 publications  were incForty-one (25.5%) of the 160 included publications reported SAEs . Only siOf the forty-one publications that reported SAEs, forty publications reported whether the SAEs resulted in death, and thirty-seven publications reported the relationship between death and the treatment, and only fifteen reported the relationship between the non-death SAEs and the anticancer treatment. The proportion of deaths caused by SAEs was as follows: less than 1% in nineteen clinical trials, 1–5% in sixteen clinical trials, and 5–10% in five clinical trials. Six publications reported whether the SAEs were life-threatening, and only two publications reported the prognosis of SAEs in detail .p = 0.003, p < 0.001). Clinical trials examining new drugs (45.5% [25/55]) liked to report SAEs more than those not examining new drugs . Clinical trials with larger sample sizes  seemed to have a greater RP of SAEs than those with medium sample sizes  and small sample sizes  and published before 2000 . The RPs of SAEs in studies published in famous journals were as follows: 25% [2/8] in N Engl J Med, 22.2% [2/9] in Lancet, 42.1% [8/19] in Lancet Oncol, 50.0% [1/2] in JAMA and 30.4% [17/56] in J Clin Oncol (p = 0.030). The RP of SAEs was significantly higher in clinical trials about second line and above treatment than those about first line and adjuvant/neoadjuvant treatment, and higher in clinical trials researched targeted therapy ± chemotherapy than those researched other therapeutic schedules  was higher than that in those conducted in local region (21.6%). The worldwide clinical research is supervised and reviewed by an international ethics committee and global regulatory agencies. The management system is stricter, so the reporting of SAEs is more stringent. In addition, clinical studies conducted in only one country had various reports of SAEs. The RP of SAEs was higher in China, Spain, Italy, and Greece, at 75, 50, 50, and 33.3%, respectively . This map < 0.001). In addition to the effectiveness of new drugs, the safety of new drugs was of paramount concern, so the RP of SAEs was higher. Non-new drug research mainly compared the efficacy of different treatment regimens and paid less attention to SAEs, and the RP of SAEs was lower. The SAEs reporting rate of clinical studies initiated by pharmaceutical companies (57.6%) was higher than that of investigators (20.7%) (p < 0.001). Among 33 clinical studies initiated by pharmaceutical companies, 69.7% (23) were new drug-related clinical studies, while only 39% (32 of 82) of clinical studies undertaken by investigators were new drug studies. The RP of SAEs in new drug clinical research was higher, so the RP of SAEs in clinical research initiated by pharmaceutical companies was higher. This was also the reason why the RP of SAEs was higher in clinical trials about second line and above treatment, and higher in targeted therapy based clinical trials.The RP of SAEs in new drug clinical research (45.5%) was significantly higher than that in non-new drug clinical studies (15.2%) . This was affected by the journal’s requirements. For example, Lancet has provided readers with links to websites that published clinical trial protocols since 2009, and J Clin Oncol has disclosed agreements that were previously only open to journal editors and reviewers since 2011   of the NSAEs reports need more improvement. For example, improving the construction of SAEs reporting systems in electronic information platforms, establishing precise process and data collection methods, strengthening the training of medical staff, enhancing the safety ability assessment of patients via patient education, and improving the awareness and attention of SAEs have been reported. The authors believe that co-report of AEs/SAEs via PRO and researchers in clinical trials should be adopted in the future .There were some shortcomings in this study. First, this was a retrospective study, and there may be omissions in data collection and selection bias. Second, the identification criteria for SAEs may vary because of the diverse designs of clinical trials and different judgment criteria of investigators. Third, bias existed in the data collection because the descriptions of SAEs in the publications were inconsistently attributed to the journal-specific publication requirements. Finally, this study’s included publications were all published clinical trials, and unpublished clinical trials, such as clinical trials with negative research results, were excluded. The reporting methods for SAEs have gradually improved as people pay increasing attention to SAEs. Independent reporting of SAEs by patients and researchers may better guide clinical practice and drug development in the future.In conclusion, our findings showed that the RP of SAEs increased and aroused more researchers’ attention over time. However, more efforts should be made to improve the RP of SAEs and the quality of SAEs reporting. The patterns and outcomes of SAEs should be reported in detail and given more attention to better guide drug application by clinicians in the real world. In addition, independent reporting of SAEs by patients and researchers should be encouraged.