Background and Objectives: Our study aimed to investigate the gross anatomy aspects of the fossa ovalis (FO) and the presence of some anatomical variation resulting from the incomplete fusion of septum primum and septum secundum, such as an atrial septal pouch (SP) and left atrial septal ridge. Materials and Methods: Thirty-one adult human hearts removed from formalin-fixed specimens were examined to provide information about the morphology of the FO. The organs were free of any gross anatomically visible pathological conditions. Results: The most common variants were the FO located in the inferior part of the interatrial septum (64.51%), circular (61.3%), with a net-like structure (51.62%), prominent limbus (93.55%), and patent foramen ovale (PFO) (25.8%). The right SP was observed in 9.67% of specimens, the left SP was observed in 29.03% of cases, and in 51.61% of cases, a double SP was observed. One sample presented a right SP and a double left SP, and one case showed a triple left SP, which was not reported previously to our knowledge. Conclusions: Knowledge of the interatrial septal anatomy becomes important for interventional cardiologists and should be documented before transeptal puncture. A true septum represents a partitioning wall shared by two structures , which cOur study supplements the knowledge of the morphology of the “true” interatrial septum, which is important in order to avoid exiting the heart during interventional cardiac procedures .-Position of the FO in the interatrial septum;-Shape of the FO;-Aspect of the FO;-Presence and location of the limbus of the FO;-Presence of the septal pouch (SP), right SP opened into the right atrium, left SP opened into the left atrium, and the position of the SP in interatrial septum;-Presence of patent foramen ovale (PFO).The study was conducted in the Department of Anatomy and Embryology, “Victor Babeş” University of Medicine and Pharmacy, Timisoara, and included 31 human hearts removed from formalin-fixed specimens. The hearts were free of any gross anatomically visible pathological conditions. The right atrium was opened by a classical incision , extendiThirty-one formalin-fixed human hearts were carefully examined, with observations below on the morphology of the FO:Regarding the position of FO in the interatrial septum, FO was positioned as inferior in 20 specimens (64.51%), anterosuperior in 2 specimens (6.45%), and in the middle of the interatrial septum in 9 specimens (29.03%).The shape of the FO  was vari(1)Six specimens (37.5%), on the entire surface of the FO;(2)Six specimens (37.5%), on the posteroinferior part of the FO;(3)Four specimens (25%), only on the inferior part of the FO.The aspect of the FO was smooth in 15 specimens (48.38%) and has a net-like structure or was cribriform in 16 specimens (51.62%). Cribriform FO presents the fibrous-strands, different from the Chiari network, described in the literature as a mobile reticulated network structure originating from the Eustachian valve, connecting to other parts of the right atrium ,11. In tIn one case , a promiConcerning the presence and location of the limbus or annulus of the FO, the limbus (annulus) was flat in 2 cases (6.45%) and prominent in 29 cases (93.55%).(1)All around the circumference in 2 cases (6.89%);(2)Superior in one case (3.44%);(3)Superior and anterior in 3 cases (10.35%);(4)Superior, posterior, and inferior in 13 cases (44.82%);(5)Anterior, superior, and posterior in 7 cases (24.14%);(6)Superior and posterior in 3 cases (10.35%).The segment of the limbus or annulus that was prominent was: The SP was bilateral (right and left double SP—(1)Right and double SP had the opening in the right atrium in 19 cases (61.29%) and the concavity was located in the inferior part , anteroinferior part , and posteroinferior part  of the circumference of the FO;(2)Left and double SP had the opening in the left atrium in 25 cases (80.64%). The concavity was located in the superior part , posteroinferior part , posterosuperior part , anterior part , posterior part , and anterosuperior part  of the circumference of the FO.A triple left SP  was founIn one case , we founPFO  was presRegarding the cribriform aspect of the FO, we found an unusual characteristic in six cases in which the entire surface of the FO had a net-like structure. The net-like appearance of the posterior and inferior parts of the FO represents multiple fenestrations in the septum primum. The net-like appearance on the entire surface of the FO represents fenestration of both the septum primum and the septum secundum. The cribriform aspect of the FO can be the consequence of incomplete overlap or atrophy of the septum primum or the septum secundum , consideRegarding the presence of a prominent ridge along the FO, this was described with a prevalence of 18.7% in pig hearts, 15% in ovine hearts , and in The limbus of FO, also called the ‘annulus ovalis’   or the ‘The atrial SP has been a much-discussed concept from the moment of its discovery. According to Krishnan and Salazar , the SP Case reports and epidemiologic results have demonstrated thrombus within the left SP and the relation to cryptogenic stroke. Initial results were mixed, with one study showing, and others not showing. an association between the left SP and cryptogenic stroke, requiring further evaluation, particularly in the younger patients suffering from a stroke. Although patients with interatrial abnormalities and stroke have lower thresholds for the induction of atrial fibrillation, this arrhythmia is rarely documented in patients with cryptogenic stroke and a PFO ,24.Later studies have demonstrated an association between the presence of the left SP and cryptogenic stroke. The left SP could thus be considered a possible site of thrombus formation. A further meta-analysis demonstrated the association between the left SP and cryptogenic stroke. The univariate analysis showed that the risk of cryptogenic stroke is higher among patients with a left SP than in cases without a left SP ,26,27,28The pathogenesis of cryptogenic stroke in patients with interatrial septal abnormalities is not entirely understood. The morphologic characteristics of the interatrial septum that best predict the risk of thromboembolism remain to be further clarified.The present recommendation is that any left SP finding should be noted during transesophageal echocardiography and thoroughly investigated, ideally with multiplane/tridimensional imaging to exclude thrombus. No clear consensus exists about whether the presence of a left SP alone in cryptogenic stroke is an indication for anticoagulation or other treatment ,30.Regarding the co-existence between the left SP and PFO, considering that both the left SP and PFO represent developmental defects in the interatrial septum (a left SP being the remnant of an incompletely fused septum primum and septum secundum and PFO is the sequel of an unfused septum primum and septum secundum), the study of Terpenning et al. on 275 cardiac CT found the prevalence of PFO in patients with a left SP to be 5.9% .Knowledge of the morphology of the FO becomes essential during cardiac surgery, especially the net-like structure of FO and the presence of the SP, which can create complications during the transseptal puncture and catheter manipulation.


A rapid and nondestructive method is greatly important for the classification of aflatoxin B1 (AFB1) concentration of single maize kernel to satisfy the ever-growing needs of consumers for food safety. A novel method for classification of AFB1 concentration of single maize kernel was developed on the basis of the near-infrared (NIR) hyperspectral imaging (1100–2000 nm). Four groups of AFB1 samples with different concentrations  and one group of control samples were prepared, which were preprocessed with Savitzky–Golay (SG) smoothing and first derivative (FD) algorithms for their raw NIR spectra. A key wavelength selection method, combining the variance and order of average spectral intensity, was proposed on the basis of pretreated spectra. Moreover, principal component analysis (PCA) was conducted to reduce the dimensionality of hyperspectral data. Finally, a classification model for AFB1 concentrations was developed through linear discriminant analysis (LDA), combined with five key wavelengths and the first three PCs. The results show that the proposed method achieved an ideal performance for classifying AFB1 concentrations in a single maize kernel with overall accuracy, with an F1-score and Kappa values of 95.56%, 0.9554, and 0.9444, respectively, as well as the test accuracy yield of 88.67% for independent validation samples. The combinations of variance and order of average spectral intensity can be used for key wavelength selection which, combined with PCA, can achieve an ideal dimensionality reduction effect for model development. The findings of this study have positive significance for the classification of AFB1 concentration of maize kernels. Maize is one of the most important food crops in the world, but it is vulnerable to fungus infection  in unsuitable storage conditions. Aflatoxin is a secondary metabolite of some species of Aspergillus fungi , which has been listed as ‘I’ carcinogen by the International Agency for Research on Cancer (IARC) due to its carcinogenicity and mutagenicity . AflatoxIn the previous decades, AFB1 was detected mainly by thin layer chromatography (TLC), high-performance liquid chromatography (HPLC), and enzyme-linked immunosorbent assay; although these methods are accurate, their instruments are complex and detection speed is slow. In other words, these methods mentioned above could not meet the needs for a rapid nondestructive method in modern industry, as they are inefficient for a large of samples.As a rapid and nondestructive technology, hyperspectral imaging technology integrates image information and spectral information into one system, which could record spatial and spectral information of given samples simultaneously ,5. At prLinear discriminant analysis (LDA) is a classic classification algorithm, which is widely used in the field of pattern recognition and image processing. Nowadays, LDA has been successfully used in the classification of hyperspectral data. Li et al.  proposedThe hyperspectral image information is huge and redundant within adjoining wavelengths; it is needed to compress data from high dimensionality to low dimensionality for improving detection efficiency using variable selection algorithms. Previous studies on this aspect often reduced the dimensionality of hyperspectral image with a dimensionality reduction algorithm, and then achieved the classification of different concentrations of AFB1 samples combined with the classification model. In this study, the dimensionality reduction in hyperspectral data was discussed, and a new key wavelength selection method was proposed by considering the between-class variance and order of average spectral intensity. A new attempt was carried out on combining this method with dimensionality reduction algorithm for compressing hyperspectral data; the result was better than that of using dimensionality reduction or characteristic wavelength alone, and the efficient compression of hyperspectral data is realized.The present study aims to investigate the feasibility of using the combination of dimensionality reduction and key wavelengths to obtain the best modeling data in the NIR region of 1100–2000 nm, combined with classification model to detect AFB1 concentration in maize kernels. The specific objectives of this study were to: (1) compare the differences between raw and pretreated spectra of different AFB1 concentrations of maize kernels and analyze the effects of pretreatment; (2) select the optimal dimensionality reduction algorithm for this study through analysis and comparison of different dimensionality reduction methods; (3) select key wavelengths according to spectral features, such as the variance and order of average spectral intensity, and identify the optimal data for modeling through the combination of specific wavelength and dimensionality reduction algorithm; and (4) obtain an optimal calibration model by comparing different classification models based on low-dimensional data.The variety of maize kernel in this study is Zhengdan 958, which is widely planted in China. All samples were provided by Beijing Academy of agriculture and Forestry Sciences, and were planted in Xiaotangshan, Beijing, North China, one of the main maize producing areas in China. Therefore, the research object of this study is representative. A total of 450 maize kernels without any defect were selected to make the maize kernel samples, with five different concentrations of AFB1. In order to ensure that there were no pre-existing natural toxins, 20 representative kernels were randomly selected from each group and the samples were tested as toxin-free by Beijing Putian Tongchuang Biotechnology Co., Ltd. In order to prepare the maize samples with five different concentrations of AFB1 artificially , five different concentrations of AFB1 diluent were prepared according to the average kernel weight (about 0.5 g).−1 diluent was created by mixing 0.25 mL of solution of toxin with 2.25 mL of methanol, and 90 samples with 10 ppb AFB1 contents were prepared by depositing 25 μL diluent on each embryoid side of each kernel (25 μL × 0.2 μg/mL = 0.005 μg of toxin per 0.5 g kernel). To achieve 20 ppb kernel samples, 0.4 μg/mL diluent was created by mixing 0.5 mL of solution of toxin with 2 mL of methanol, and 25 μL of diluent was deposited on each kernel to achieve 20 ppb (25 μL × 0.4 μg/mL = 0.01 μg of toxin per 0.5 g kernel). Following a similar procedure, different concentrations of diluents were prepared by changing the mixture ratio of toxin and methanol. Then, the diluents of different concentrations were dropped on the kernels to achieve 50 and 100 ppb. Each sample in the control group was treated with a 25 μL methanol-alone drop to eliminate the effect of diluent treatment as much as possible. After these kernels were inoculated, they were placed in a chemical hood to allow the diluent to dry. Then, after 120 min, the methanol in these samples volatilized, and AFB1 was absorbed completely by the samples [The diluent and samples were prepared as follows: initially, 5 mL of AFB1 toxin (2 μg/mL), 5 mL of 100% methanol, and other necessary equipment were prepared. Then, 0.2 μg ml samples ,18,19. AA line-scanning NIR hyperspectral imaging system  was usedIn order to acquire the high-quality hyperspectral images, the distance between the lens and the sample stage was set at 380 mm, and two halogen light area sources were mounted at 45° angles from horizontal to avoid oversaturation. The movement speed of the sample and the exposure time of camera were set at 42 mm/s and 2 ms, respectively, to avoid image spatial deformation. In order to improve the image collection efficiency, 90 samples with the same AFB1 concentrations (15 rows and 6 columns) were placed on the transport platform and scanned line by line each time. Each NIR hyperspectral image consists of 256 congruent subimages, with wavelengths ranging from 930 nm to 2548 nm. Due to the severe noise in the spectrum before 1100 nm and after 2000 nm, only the spectral range covering 1100–2000 nm (145 wavelengths) was used for the NIR imaging modes to minimize the effect of noise [The spatial intensity variation in the plane of the scene produced by halogen lamps and the dark current in the CCD camera generally led to the signal of the camera chip to be nonzero when no light hit the detectors and resulted in a large amount of noise in some bands . TherefoIn Equation (1), A hyperspectral image consists of image information and spectra information. In order to develop a maize kernel classification model with different concentrations of AFB1, the image and spectra information of maize kernel were extracted as follows:Step 1. The grayscale image at 1210.3 nm was selected to segment the maize kernel from the background due to their obvious intensity difference;Step 2. Threshold segmentation and morphological filtering were employed to remove the background and edge region of the maize samples while remaining in the effective region at the grayscale image at 1210.3 nm;Step 3. The filtered image was used as the region of interest (ROI) of the sample to mask the original hyperspectral image and extracted the region of each kernel;Step 4. The spectra data of each kernel within the ROI were averaged at each wavelength, and the average spectra were used for further analysis.The hyperspectral data also contained some disturbances  during the NIR spectra acquisition. In order to reduce the influence of nontarget factors and to establish a stable model, the raw spectra must be pretreated before further analysis . SavitzkVariable selection methods are usually used to select the key wavelength. Variance could reflect the differences between groups; in this study the key wavelengths used for the classification of AFB1 content in maize kernels were determined by combining the variance of spectral intensity with their order. The specific method is as follows:Step 1. Calculated the average spectrum of each group;Step 2. Ordered the average spectral intensity of five groups at each band, and divided the bands with the same order into the same subset. In order to eliminate the effect of tiny intensity difference on ordering, the spectral intensity of adjacent groups would be regarded as equal if their difference was less than 100 times of variance;Step 3. Calculated the variance of spectral intensity for each band, and only the wavelength with the largest variance was retained in the same subset; the remaining wavelengths were removed;Step 4. All the remaining wavelengths were sorted according to the variance from large to small, and the first few retained wavelengths were the final selected characteristic wavelengths.The flowchart of key wavelength selection in this study is shown in PCA is a well-known algorithm of multivariate analysis . The maiThe basic idea of LDA is to compress the dimensionality of feature space and extract the classification information through projecting the high-dimensional data into the optimal discriminant vector space. The projected data have the minimum within-class distance and the maximum between-class distance in the new subspace, that is, the pattern has the best separability in the new vector space .For a given data set In Equations (2) and (3), According to the nature of the ‘Generalised Rayleigh Quotient’, the optimal classification vector space obtained ,30. ThenIn this study, there were four groups of samples with different AFB1 contents and one control group of samples without AFB1. All 450 samples (five groups with 90 samples in each group) were randomly divided into the calibration set or prediction set, according to the ratio of 1:1. Thus, 225 samples (45 of each group) were selected as the calibration set to develop the calibration model, and the remaining 225 samples (45 of each group) were used as the prediction set to verify the performance of the developed model. To eliminate the influence of random sample division on the prediction ability of the developed model, the prediction model was developed 10 times by random sample division; the model which came closest to the average performance of the 10 developed models was taken as the final model. To evaluate the classification performance of the proposed method, three parameters, namely accuracy, F1-score, and Kappa, were considered ,32. The The raw and pretreated spectra of all samples in the calibration set are presented in 0, V10, V20, V50, and V100 represent the average spectral values when AFB1 concentration is 0, 10, 20, 50, and 100 ppb at the selected wavelength, respectively. The variance at the first five bands were 4.37, 1.89, 1.83, 1.12, and 1.06, respectively, but the variance decreases rapidly from the sixth wavelength, therefore only the first five wavelengths  were selected as key wavelengths according to our proposed method , factor analysis (FA), t-distributed stochastic neighbor embedding (t-SNE), and random projection (RP), were selected. It can be seen from To highlight the superiority of LDA classifier in this study, three other classifiers, including K-nearest neighbors (KNN), Naive Bayes (NB), and Decision Tree (DT), were used to establish the prediction model based on the same dataset, and then their classifier result was compared with LDA . The disTo further verify the effect of this model, 30 new samples in each group were re-prepared with the same method. The hyperspectral images of samples were preprocessed in the same way. Under the LDA model, the spectral data of the five key wavelengths and the first three PCs combinations were used for testing, and the results are shown in It can be seen from Using NIR spectroscopy technology to discriminate the AFB1 concentration of single maize is still in the exploratory stage. It is difficult to control the content of AFB1 produced by natural mold, therefore the artificial inoculation of toxin becomes a simple and effective sample preparation method due to the surface distribution characteristics of AFB1 . Tao et An ideal result for classification of different AFB1 concentration was obtained based on the proposed method of this study. Although the established model has achieved satisfactory results, the success rate for classification of 20 and 50 ppb groups still needs to be improved. PCA has good dimensionality reduction effect. It can be seen from On the basis of the NIR hyperspectral image in the 1100–2000 nm, with pretreatment by SG and FD, five key wavelengths were selected by combining variance and order of spectral intensity, and the data of the first three dimensions were obtained by PCA dimensionality reduction based on full wavelength. Then, LDA was used to establish the classification model based on the data of combinations of five key wavelengths and the first three PCs. The proposed algorithm achieved an ideal result to classify the AFB1 concentrations of maize kernels. Four groups of samples with different AFB1 contents and a group of control samples were selected for this study; 90 samples of each group were divided into the calibration set and the prediction set, according to the ratio of 1:1. The results show that the average test accuracy of AFB1 concentrations of maize kernels in the prediction set was 95.56%, which is better than the results of full wavelengths and other dimensionality reduction methods. For independent validation samples, the test accuracy reached 88.67%. This study has certain reference value for the classification of AFB1 concentrations of single maize kernels. In the future work, different varieties of maize kernels and more accurate detection of AFB1 concentration levels will be studied.


No significant difference was observed for the intake of fruit, vegetables, or F&V. Basing food education strategies on the stages of change model shows positive results, increasing the awareness and disposition of people with SMD towards the intake of F&V. More research is needed to identify the most appropriate eating intervention to increase the intake of F&V.Despite growing evidence of the benefits of adequate intake of fruit and vegetables (F&V) and the recommendation to consume five servings daily, the adoption of these habits is poor among people with severe mental disorder (SMD). The main aim of the present study is to determine changes in the intake of F&V and motivation to do so among people with SMDs after participating in a food education programme. A community-based randomized controlled trial was conducted in Spain, with the intervention group (IG) participating in a food education programme based on the stages of change model to promote consumption of F&V and the control group (CG) receiving three informative sessions on basic healthy eating. The main outcomes were related to the intake of F&V and stages of change. Data collection was performed at baseline, post intervention, and 12-month follow-up. Seventy-four participants enrolled in the study and sixty completed the 12-month follow-up. An increase in motivation towards the intake of F&V was observed in the IG but not in the CG (McNemar’s test  Following a diet rich in fruit and vegetables (F&V) is associated with numerous health benefits ,2, not oThe World Health Organization (WHO) recommends a minimum intake of 400 g of F&V a day as part of a healthy diet, prioritizing the consumption of fresh fruit and emphasizing the importance of variety . These rIn Catalonia, only 14% of people over the age of 15 consume five or more servings of F&V a day . A low iSMDs encompass a set of mental disorders of different natures and clinical presentations, displaying common criteria in terms of severity and persistence over time, with a tendency towards deterioration and alteration of social, family, and work functioning, thus, reducing the individual’s quality of life .The adoption of unbalanced eating habits is considered to be one of the individual risk factors in the accumulation of metabolic disorders among people with SMDs, such as obesity, diabetes mellitus, and metabolic syndrome, resulting in a high presence of cardiovascular comorbidity ,17. CardHowever, the adoption of unhealthy eating behaviours is identified as a modifiable risk factor, which makes it an opportunity for intervention . DevelopThe transtheoretical model  views beSeveral studies have recognized the relevance of basing interventions on the stages of change model, allowing for strategies to be designed that respond to the needs of participants, and thus effectively facilitating the adoption of healthy behaviour ,24. AlthThe main aim of the present study is to investigate changes in the intake of F&V and motivation to do so among people with SMDs after participating in a food education programme based on the stages of change model. In addition, we also investigate the association between the intake of F&V and non-motivational factors  and describes the disposition towards the intake of five daily rations of F&V according to the aforementioned model.n = 74) consisted of individuals with SMDs who were also participants at the Osonament community mental health services, a comprehensive care service offering a network of resources to cover needs related to recovery, housing, leisure, and employment for people with mental health problems and addictions in the region of Osona .A randomized community-based controlled trial was conducted between January 2019 and September 2020. The sample  or CG using the Zenon algorithm (equiprobable randomization 1:1 through R Software), considering the variables of age, gender, functionality, and primary mental health diagnosis. A researcher otherwise unconnected to the study performed the randomization.All participants, and legal guardians if required, gave their consent for inclusion before participating in the study. The study was conducted in accordance with the Declaration of Helsinki, and the protocol was approved by the Clinical Research Ethics Committee (CEIC) belonging to the Osona Foundation for Health Research and Education (FORES) in September 2018 (CEIC code 2018974).The Dietment intervention programme lasted four months (April to July 2019). It consisted of a food education strategy aimed at promoting the consumption of F&V and comprised 15 group sessions (of 5–10 people) lasting 90 min each session, which were held weekly. All sessions were conducted by the same dietitian-nutritionist.The intervention design was based on the stages of change construct included in the transtheoretical model . ConsideThe food recommendations provided were taken from the guide to healthy eating prepared by the Catalan Public Health Agency . No foodData were collected at baseline (T0) and once completed the intervention (T1) for both groups (IG and CG). To determine the long-term effect, it was repeated 12 months after the end of the intervention (T2). A questionnaire was used to re-collect the sociodemographic and clinical data as well as the different study variables.Sociodemographic variables included age, gender, marital status, level of education, and whether support was available for carrying out activities of daily living (ADL).The mental health diagnosis was obtained from the medical history. At the anthropometric level, the current weight and height were collected, making it possible to obtain the body mass index (BMI) . The FunData on the consumption of F&V were collected by means of two questions related to: “serving of fruit consumed the day before” and “serving of vegetables consumed the day before”. The answers to the two questions were added together to obtain the total consumption of F&V. The interviewer explained which foods were included in the two groups, as well as what constituted the consumption of a serving of F&V.Classification into each of the stages was determined using the algorithm proposed by Laforge et al. . This isAs described by the authors, participants were asked about the number of servings consumed the day before: if the answer was five or more daily servings, they were asked if the behaviour had been maintained over more than six months. Participants who responded from 0 to 4 servings were asked about the intention to eat a minimum of five servings a day in the next six months. If they responded “yes”, they were asked whether they intended to eat a minimum of five servings per day over the next 30 days.Respondents were assigned to the precontemplation stage if they reported usually eating fewer than five servings of F&V daily and had no intention of changing their behaviour. The contemplation stage was defined as usually eating fewer than five daily servings and reporting the intention to adopt the change within the next six months. People were assigned to the preparation stage if they reported intending to adopt the change within the next 30 days. People were considered to be in the action stage if they reported usually eating five or more servings of F&V but had been doing so for less than six months. The maintenance stage was defined as eating a minimum of five servings daily for more than six months.Participants’ disposition towards switching to the consumption of F&V was not taken into account for recruitment or randomization purposes.For the analysis of paired data, the stages of change variable were re-coded to a dichotomous variable grouping the stages of pre-contemplation, contemplation and preparation (pre-action) and action and maintenance (action/maintenance).A descriptive study was carried out, the categorical variables being described using absolute frequencies and percentages, and the measures of central tendency  and dispersion (standard deviation (SD), percentiles) being calculated for the quantitative variables. The assumption of normality of the continuous variables was studied, as well as the total scores of the tests used.t-test.An analysis of independent samples was performed between the two randomization groups to study whether the groups were homogeneous a prior (baseline). The comparison for categorical variables was performed using Pearson’s chi-square test or Fisher’s test if appropriate. The comparison of means was carried out by means of a Student’s The Pearson’s correlation coefficient, Anova, and Student’s t-test were used to analyse the association between consumption of F&V and the different sociodemographic and clinical variables. The Anova and post hoc tests were used to determine the association between the stages of change and the intake of F&V.Given the small size of the study sample, non-parametric techniques were applied (Friedman and Wilcoxon signed-rank test) to study the evolution in the number of rations of F&V consumed between baseline, post, and 12-month follow-up evaluations and for the two intervention groups. The Mann–Whitney U Test was applied to detect a difference between groups at baseline (T0), post intervention (T1), and 12-month post intervention (T2). The McNemar test was applied to determine the effect of the intervention on disposition towards change, based on the comparison of response percentages prior to and after the intervention.Seventy-four users of the Osonament community services completed baseline data collection, 37 of whom were assigned to the IG and 37 to the CG. The original target sample size could not be reached. Complete data could be collected for 67 people at the end of the intervention (T1), and for 60 people one year after its completion (T2). The mean age was 48.7 years (SD 10.8), the majority (55.4%) were men. The most common diagnosis of mental health was schizophrenia and other psychotic disorders (37.8%). At baseline, the groups were homogeneous in terms of sociodemographic and clinical characteristics . Althougn = 74). No statistically significant associations were found with age, gender, marital status, level of education, primary diagnosis of mental health, weight, BMI, functionality, cognitive impairment, or subjective well-being. A significant association was only observed between the average serving of F&V consumed and support received for ADL: 3.2 (SD:1.5) in individuals receiving professional support, 1.9 (SD:1.7) in those not receiving support, and 1.8 (SD:1.6) in those receiving family support (p = 0.022).n = 74). Some 63.5% of participants were in pre-contemplative or contemplative stages, and 10.8% reported following the defined behaviour (action and maintenance). p < 0.001). Those participants in the action/maintenance stages reported a significantly higher intake of F&V than those in the pre-contemplation and contemplation stages.n = 74), 10.7% of participants reported consuming the recommended amount of F&V (five servings/day) (5.4% of the IG participants and 16.2% of the CG participants), the median number of servings consumed being two (IQR 2.3). Some 82.4% ate fewer than three servings of fruit and 82.4% ate fewer than two servings of vegetables. At post intervention (n = 67), 15% of participants achieved five daily servings of F&V (21.3% of the IG participants and 8.8% of the CG participants).At baseline (p = 0.124), vegetables (p = 0.110), or F&V (p = 0.595) between the IG and the CG at baseline (n = 74). The IG showed a significance increase in the consumption of fruit and F&V over time , but no difference was observed in the CG (p = 0.086). No differences were observed in relation to vegetable intake in the two randomized groups over time , while the change was not significant for the CG (p = 0.625).Between baseline and post intervention, 23.3% of the IG participants progressed to the action/maintenance stage , no significant differences were observed in either of the groups. A dietary education intervention based on the stages of change model to promote consumption of F&V among people with SMDs was found to increase the number of people who switch to action/maintenance stages by 23%. Despite the observation of an increase in fruit intake over time by the IG, the change was not significant as compared with that of the CG. This study did not find any significant difference in the intake of vegetables and F&V between the groups over time, highlighting the challenges of promoting healthy eating habits among people with SMD.At baseline, only 10.7% of the sample studied achieved the recommended intake of F&V, with an overall average consumption far below the five recommended daily servings of F&V. Various studies have shown the consumption of F&V to be below the established recommendations among people with mental disorders . In a SpThere are many factors that can condition and influence food choices . AlthougOther authors have identified a relationship between gender, age, and BMI and the intake of F&V . In a quOur study indicates that at baseline almost half of the participants (47%) received professional or family support to carry out basic self-care activities and instrumental activities independently. However, it should be noted that such support does not always encourage healthy eating habits, as the results show the consumption of F&V to be below the minimum recommended intake. Thus, receiving support is identified as a favourable aspect in the adoption and maintenance of healthy eating behaviours, with a positive impact on the quality of the diet, as long as this environment promotes health and the people who are responsible for eating follow a healthy eating pattern themselves . With regard to the disposition of the sample studied towards increasing the intake of F&V, most participants were in stages of non-disposition; 64% reported being pre-contemplative or contemplative at baseline, denoting a low awareness of the consequences of insufficient intake of F&V, while those in the pre-action stages were found to display a lower intake of F&V as compared with those in the action stages. On the basis of a qualitative exploration of barriers to adopting healthy eating in adults with mental health problems, Barre et al. observed that most were in stages of pre-contemplation or contemplation, and that those who adopted dietary changes towards a healthier eating pattern were motivated by a perception of personal risk after an episode of illness . StudiesTo date, several authors have designed and implemented interventions to promote healthy lifestyles aimed at people with mental disorders . DespiteOur results show that using the stages of change model to design interventions for promoting healthy eating aimed at people with SMDs returns favourable results with regard to increasing motivation for the consumption of F&V. Basing the intervention design on the transtheoretical model allows for the use of specific intervention strategies that increase in effectiveness with each stage, providing a series of defined strategies to facilitate movement towards stages of greater disposition towards the desired behaviour . AlthougLimitations of the present study include: the small sample size may limit the power of the study in detecting differences between groups, while it may also condition uneven sampling at different stages of change; collecting information on the intake of F&V that is only based on consumption in the previous 24 h can present a bias in determining actual intake—a prospective 72-hour record or a record of non-consecutive days could also be included; there is a possible risk of information contamination between the participants of the IG and the CG, due to their attending the same centre and being located in the same environment; and finally, the period between post intervention and 12-month follow-up coincided with the COVID-19 pandemic, although there is insufficient evidence of this exerting a possible influence on the results obtained.The strengths of the study include its design, which allowed for comparison with a control group, and follow-up of participants at 12 months post intervention. In addition, the inclusion of programmes for promoting healthy eating habits in community rehabilitation services as an integral part of the individual’s recovery process, and the evolution of individuals’ disposition towards acquiring healthy eating behaviours. Although the sample size can be considered small at the statistical level, taking into account the target profile, it is concluded that there has been a high participation and adherence to the programme, with less than 20% loss one year after the end of the intervention.Despite growing evidence of the benefits of an adequate intake of F&V, people with SMDs who participated in this study displayed a consumption that was far removed from the established recommendations of five daily servings. This highlights the need to develop food education programmes that facilitate the disposition towards changing behaviour and promoting the intake of F&V among individuals in these groups. Despite there being a trend to increase fruit intake among the IG, this result was not statistically significant as compared with the CG. Basing food education strategies on the stages of change model yields positive results in relation to an increase in the disposition of people with SMDs towards a higher intake of F&V. More experimental studies are needed on the effectiveness of behavioural interventions aimed at changing eating habits in people with SMDs.


The aim of the analysis was for the first time to assess the expression of genes encoding IL-21 and IL-22 at the mRNA level in ovarian tumor specimens and the concentration of these parameters in serum and peritoneal fluid in patients with ovarian serous cancer. The levels of IL-21 and IL-22 transcripts were evaluated with the use of the real-time RT-qPCR. Enzyme-linked immunosorbent assay (ELISA) was used to determine the concentration of proteins. Quantitative analysis of IL-21 gene mRNA in the tumor tissue showed the highest activity in the G1 degree of histopathological differentiation and was higher in G1 compared to the control group. The concentration of IL-21 and IL-22 in the serum and in the peritoneal fluid of women with ovarian cancer varied depending on the degree of histopathological differentiation of the cancer and showed statistical variability compared to controls. The conducted studies have shown that the local and systemic changes in the immune system involving IL-21 and IL-22 indicate the participation of these parameters in the pathogenesis of ovarian cancer, and modulation in the IL-21/IL-22 system may prove useful in the development of new diagnostic and therapeutic strategies used in patients, which require further research. Ovarian cancer is one of the gynecological cancers with the worst prognosis. Both the number of cases and the mortality caused by this disease in the world are constantly increasing ,2. In paRecent studies indicate that interleukin 21 (IL-21) and interleukin 22 (IL-22) play an important role in the pathogenesis of cancer and cancer therapy ,10,11,12Interleukin-22 is a cytokine synthesized by lymphocyte T helper (Th) cells, including their Th17 and Th22 subpopulations as well as NKT lymphocytes, NK cells, and nonspecific lymphoid cells . It is aThe analyses conducted so far on the evaluation of IL-21 and IL-22 in ovarian cancer have been interesting and induce further research. It has been shown that the determination of IL-21 with other cytokines, for example, IL-17a, may find clinical application as prognostic and therapeutic biomarkers . InterleSo far, no studies have been conducted to assess both the expression of IL-21 and IL-22 in tumor tissue, as well as the concentration of these cytokines in the serum and peritoneal fluid in women with ovarian cancer, and considering the fact that understanding the role of these parameters in the pathogenesis of ovarian cancer may be useful in developing new diagnostic and therapeutic regimens in patients with ovarian cancer in the future, the aim of the study was: assessment of IL-21 and IL-22 at the mRNA level in ovarian cancer tissue, analysis of the concentration of IL-21 and IL-22 in the serum and peritoneal fluid of patients with ovarian cancer, and determining whether there is a relationship between the concentration of IL-21 and IL-22 and the degree of histological differentiation of the cancer.g for 15 min. The serum obtained in this manner was kept in small portions at a temperature of −80 °C until the tests. Tumor tissue intended for molecular assessment was collected during the planned surgery and frozen at −80 ° C until the tests were performed. Peritoneal fluid was collected during laparoscopy for bacteriological examination, and then centrifuged at 2000 rpm for 10 min at 4 °C, and the obtained supernatant was partitioned and frozen at −80 °C until the remaining determinations were made.The study group included 26 women, aged 41 to 82 (mean age: 62.80 ± 11.21 years) with the diagnosed ovarian cancer with the Cystadenocarcinoma papillare serosum IIIC  Staging employed the criteria recommended by the International Federation of Gynecology and Obstetrics (FIGO). The diagnosis of tumors was done on the basis of clinical symptoms, results of gynecological and histopathological examination and laboratory tests. The women qualified to the studied group were clinically diagnosed with ovarian tumor confirmed with a histopathological examination. Other coexisting disorders of the reproductive organs and autoimmune diseases were excluded. None of the examined women used pharmacological treatment in the last three months. The research was conducted on women hospitalized at the Clinical Department of Gynecology and Obstetrics of the University Clinical Center in Katowice and the Department of Gynecology and Obstetrics with the Department of Pathology of Pregnancy and Gynecological Oncology of the Provincial Specialist Hospital in Częstochowa. Serum, peritoneal fluid, and tumor tissue were the material examined in all women. In the study group, blood was taken from women after establishing clinical diagnosis, before surgery. Blood was taken in the morning from the cubital vein, to a clot tube, in order to obtain the serum. Thirty minutes after taking the blood, it was centrifuged at 1500× The control group consisted of six women aged between 40–77 (mean age: 61.83 ± 12.48 years) who have been diagnosed with a benign lesion (Cystadenoma serosum). The concentration of antigen CA125 did not exceed 35 U/m in these patients. In all the women, the blood serum and tissue samples were the research material.® homogenizer . RNA extracts quantitative assessment was performed with the use of nanospectrophotometer MaestroNano MN-913  and 2100 Bioanalyzer   and GoTaq® 1-Step RT-qPCR System , according to manufacturers’ instructions. Amplification was performed using previously described oligonucleotide primers [Total RNA was extracted from tissue samples with the use of TRIzol reagent , according to the manufacturer’s instructions. Each tissue sample was placed in a tube containing TRIzol, and then homogenized using a PolytronCA, USA) . The lev primers ,24 and cAll the women who participated in the study consented to conducting the research. The approval of the Ethics Committee of the Medical University of Silesia in Katowice was obtained.r).The obtained results were statistically analyzed using the Statistica 13.3 software . The normality of the distribution of the studied variables was checked using the Shapiro–Wilk test. The median and quartile range were determined for the parameters tested, and the obtained results were compared using the Mann–Whitney test. Correlations were tested by Spearman’s rank correlation test and presented as correlation coefficient (p = 0.002415), G2 (p = 0.033161), and G3 (p = 0.020397). No statistically significant difference was found between G2, G3, and a control group. Moreover, expression of IL21 mRNA did not differ between G2 and G3.Quantitative assessment of IL21 mRNA in tissue samples showed the highest gene activity in the degree of differentiation G1 . The expQuantitative assessment of IL22 mRNA in tissue samples did not show statistically significant difference comparing to a control group. Moreover, expression of IL22 mRNA did not differ between degrees of differentiation G1, G2, and G3. Median values of the number of copies of IL21 mRNA/µg of total RNA in tissue samples were as follows: G1: Me = 6660, G2: Me = 5065, G3: Me = 7820, C: Me = 8555.p = 0.003405; G2 vs. C p = 0.008114; G1 vs. C p = 0.007473). Moreover, statistically significant difference was found between G1 and G3 (p = 0.032391).Il-21/IL-22 mRNA ratio was the highest in the degree of differentiation G1 . In all The concentration of Il-21 was determined in the serum and peritoneal fluid of women with ovarian cancer. As the obtained values did not correspond to the normal distribution, the results were presented in the form of the median and the lower and upper interquartile range (Q1 and Q3).p < 0.001), where Q1 and Q3 were respectively: 722.19 and 909.60 with a median of 793.59 pg/mL.In the serum of women with ovarian cancer, Q1 and Q3 were respectively: 314.25 and 736.00 with a median of 408.68 pg/mL. A statistically significantly lower concentration of IL-21 in the serum of women with ovarian cancer was demonstrated compared to the concentration in the control group (p < 0.0001).In the peritoneal fluid of women with ovarian cancer, Q1 and Q3 were: 113.36 and 356.13, respectively, with a median of 175.63 pg/mL. A statistically significant higher concentration of the examined parameter was found in the serum of women with ovarian cancer as compared to the concentration in the peritoneal fluid (p < 0.001) and between G1 and G3 (p < 0.0001) in the serum of the examined women. In the peritoneal fluid of women with ovarian cancer, a statistically significant difference was found only between G2 and G3 (p < 0.05). The obtained results are presented in Then, the concentration of IL-21 in serum and the peritoneal fluid of women with ovarian cancer depending on the degree of histopathological differentiation of the cancer was analyzed. The analysis of the test results showed a statistical significance between G2 and G3 .p < 0.001), where Q1 and Q3 were respectively: 66.01 and 140.04 with a median of 98.62 pg/mL.In the serum of women with ovarian cancer, Q1 and Q3 were respectively: 337.59 and 771.33 with a median of 572.64 pg/mL. A statistically significant higher concentration of IL-22 in the serum of women with ovarian cancer was demonstrated compared to the concentration in the control group (p < 0.0001).In the peritoneal fluid of women with ovarian cancer, Q1 and Q3 were 132.40 and 275.70, respectively, with a median of 186.01 pg/mL. A statistically significant higher concentration of the examined parameter in the serum of women with ovarian cancer was demonstrated compared to the concentration in the peritoneal fluid (p < 0.0001) and between G1 and G3 (p < 0.0001) in the serum of women. In the peritoneal fluid of women with ovarian cancer, no statistically significant differences were found between the degrees of histopathological differentiation of the cancer.Then, the concentration of IL-22 in serum and peritoneal fluid in women with ovarian cancer depending on the degree of histopathological differentiation of the cancer was analyzed. The analysis of the test results showed a statistical significance between G2 and G3 . The studies showed that in the samples of the G1 histological differentiation, the number of mRNA copies of IL21 gene was statistically significantly higher, not only compared to the control group, but also compared to the G2 and G3 histological differentiation samples. In the case of IL22, we did not observe any differences in the number of mRNA copies between the cancer specimens and the control group, as well as between the G1, G2, and G3 histologically differentiated groups. The studies by Fagerberg et al.  did not Our studies showed a statistically significantly lower concentration of IL-21 in the serum of women with ovarian cancer compared to the concentration in the control group, which indicates the participation of the tested cytokine in the immune response against ovarian cancer cells. In addition, statistical significance was demonstrated between the concentration of the tested cytokine and the degrees of histological differentiation in cancer: G2 and G3, and between G1 and G3, which indicates a relationship between IL-21 secretion and histological differentiation in cancer and may prove useful in the future in selecting the optimal therapeutic treatment and in the assessment of prognosis.Interesting observations were also provided by the analysis of the concentration of IL-21 in the peritoneal fluid of the examined women. The concentration of this parameter was significantly lower in the peritoneal fluid as compared to the serum, which may indicate that the source of the tested cytokine in the serum may be cells of the immune system, which, through secreted mediators, show increased cytotoxic activity against cancer cells. Moreover, it was shown that the concentration of IL-21 in the peritoneal fluid of women with ovarian cancer differed between the grades of G2 and G3, which proves that the secretion of IL-21 is related to the histological differentiation of ovarian cancer also in this biological fluid.So far, no studies have been conducted to evaluate the expression of IL-21 in tumor tissue and the concentration in serum and peritoneal fluid. However, the potential usefulness of using this cytokine in the diagnosis of ovarian cancer was analyzed. Such research was conducted by Chen YL. et al. , who shoThe increase of IL-21 mRNA in tumor tissue at the G1 stage may indicate that there are still efficient mechanisms aimed at limiting its growth, which is also reflected in the IL21/IL22 mRNA ratio value. In the G2 and G3 stages, the expression is reduced in tumor tissue, possibly as a result of disturbed intracellular signaling or epigenetic changes.On the other hand, the increase in protein concentration, both in the serum and in the peritoneal fluid, suggests the activation of systemic mechanisms and a strong recruitment of the immune system. Due to blocked mechanisms in the tumor tissue itself, the antitumor effect of the immune system is probably ineffective, despite its strong stimulation. However, this hypothesis requires further investigation. It should also be taken into account that gene expression is a multi-step process and its regulation is complex. Strongly activated gene transcription in the tumor tissue does not mean that protein is synthesized and secreted. In our research, the protein source are likely cells of the immune system. If tumor tissue were the main source of interleukins, the protein concentration would be higher in the peritoneal fluid than in the serum.An interesting observation is the fact that serum protein concentration is significantly decreased by the G1 stage and the concentration increases with the degree of histological differentiation of the cancer. In the G3 stage, the IL21 concentration increases to a level comparable to that in the control. Most likely, in the lower degree of histological differentiation of the cancer, the mechanisms responsible for “masking” the presence of cancer cells are activated. In our research, the study group was not very large, so the results should be interpreted with caution.Further analysis included the evaluation of interleukin 22 expression at the mRNA level in tumor tissue and serum and peritoneal fluid concentration in women with ovarian cancer. The studies conducted so far have shown that the role of the interleukin IL-22 is complex. On the one hand, this cytokine promotes tumor development, is responsible in particular for the proliferation of malignant neoplasm of epithelial origin, but on the other hand, the intensification of its secretion may constitute a barrier against tumor development by suppressing inflammation due to the functions that protect against tissue damage, promote tissue repair, and prevent inflammation ,34. Our Further analysis concerned the assessment of the relationship between the concentration of IL-21 and IL-22 in the serum and in the peritoneal fluid. The demonstrated correlation between the concentrations of the tested parameters, both in the serum and in the peritoneal fluid, may indicate the need for further research in order to create complex diagnostic and therapeutic strategies targeting many different mechanisms of the immune response. Due to the fact that the interactions between these parameters may affect the direction of their biological activity, the stoichiometric relationship between the concentration of IL-21 and IL-22 and the number of IL21/IL22 mRNA copies was assessed. The determined IL-21/IL-22 ratio, both at the mRNA level in the tissue and at the protein level in the serum and in the peritoneal fluid in patients with ovarian cancer, showed a variable share of the tested cytokines in the IL-21/IL-22 system, depending on the histological differentiation of the cancer.Similar studies were conducted by Balint et al. , who assLocal and systemic changes in the immune system with the participation of soluble mediators IL-21 and IL-22 indicate the participation of these parameters in the development of ovarian cancer and may participate in pro and anti-inflammatory activation involving the tested cytokines.Modulation in the IL-21 and IL-22 system may affect the course of the inflammatory process that accompanies the development of cancer, which may prove useful in the development of new diagnostic and therapeutic strategies used in patients with ovarian cancer, which requires further studies.


To investigate the incidence and characteristics of retinopathy of prematurity (ROP) premature infants with late gestational age (GA) and large birth weight (BW) and show a 7-year trend of ROP incidence in South China.This retrospective, cross-sectional study included premature infants who received ROP screening in a 7-year period (from 2010 to 2016) at the Sun Yat-sen Memorial Hospital (SYSMH), Guangzhou, South China. Infants were screened if they had GA <37 weeks or BW <2,500 g. All screened infants were divided into two groups: Group 1  and Group 2 (others). The characteristics of ROP infants in Group 1 were analyzed and compared with those in Group 2.p = 0.50) and Type 1 ROP  in Group 1 were comparable with those in Group 2. Lower proportions of respiratory distress , blood transfusion , and oxygen administration  among ROP patients in Group 1 than those in Group 2 were revealed. Vaginal delivery [OR: 4.73 (1.83–12.26)] was identified as a factor associated with ROP among the infants in Group 1. Forty percent (6/15) of Type 1 ROP in this study would have been missed under the current screening criteria in China . Trends of increased incidence of Type 1 ROP and decreased BW were exhibited in the 7-year study period.A total of 911 premature infants were screened, with 282 infants in Group 1 and 629 in Group 2. Both the incidences of any ROP (6.7 vs. 8.3%, These findings indicate that even the premature infants with late GA and large BW also have a high risk of developing ROP, especially for those delivered by vagina. The findings may provide a significant reference for ROP screening and neonatal care in South China and other regions with similar conditions. Retinopathy of prematurity (ROP) is one of the leading causes of potentially preventable and treatable blindness among premature infants worldwide . AlthougFurthermore, the Chinese Ministry of Health first issued guidelines on supplemental oxygen delivery policies and the prevention and treatment of ROP in 2004 . The guiThis was a retrospective, cross-sectional study of the incidence and characteristics of ROP among premature infants over a 7-year period (from 1 January 2010 to 31 December 2016) at the Sun Yat-sen Memorial Hospital (SYSMH), one of the largest and the oldest tertiary general hospitals in Guangzhou, South China. In the database of the Medical Records Department of SYSMH, premature infants and ROP were encoded using the International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM)  and the According to the “Chinese Ministry of Health guidelines on oxygenation policies and prevention and treatment of ROP” [versions of 2004  and 2013The first examination was performed at 32 weeks of postmenstrual age or 4–6 weeks after birth, whichever came last. If ROP-related change was not detected at the first time point, these infants were followed up every 2 weeks until the vascularization of peripheral retina was completed. Eyes were diagnosed as Type 1 ROP if they had zone 1 ROP with plus disease; zone 1, stage 3 ROP without plus disease; or zone 2, stage 2 or 3 ROP with plus disease . Once TyAll fundus examinations were performed by two experienced ophthalmologists (XG and YYL) and mutually checked once ROP was detected. Pupils were dilated with 0.5% Compound Tropicamide Eye Drops  before examination . Infants were in the supine position and were held by an experienced nurse. Binocular indirect ophthalmoscopy was performed using a lid speculum, pre-set lens (+20 Dioptres) and a scleral compressor after topical anesthesia . Infants with ROP were classified and recorded according to the International Classification of ROP.The number of premature infants and infants with ROP in the 7-year period were extracted. The disease information, including the stage, zone, presence or absence of plus disease, and laterality of patients with ROP, were collected. Potential risk factors of ROP were also extracted, including sex, number of fetuses, GA , BW, delivery manner , blood transfusion, respiratory distress, and information on oxygen administration (mode of delivery and duration of oxygen administration).To explore the appropriate screening criteria of ROP in South China, we applied the ROP screening guidelines of the US  , UK GA , and Chin) and relative frequency (%) were used for quantitative variables, such as the incidence of ROP and the ratio of males to females. An independent t-test was used to compare GA and BW between the two groups. The Pearson chi-square test or Fisher's exact test (expected count <5) were used to compare the proportions of ROP and potential risk factors between the two groups. Binary logistic regression was used to analyze the relationships between ROP and the following factors among premature infants in Group 1, including the sex, GA, BW, multiple birth, delivery manner, blood transfusion, respiratory distress, and information of oxygen administration. The odds ratio (OR) and 95% confidential interval (CI) were used to determine the risk factors for ROP in Group 1. All tests of hypotheses were 2-tailed. The level of significance was set at P < 0.05.All of the data were entered into Microsoft Excel  spreadsheets, sorted and analyzed by two researchers. The data were further imported into the Statistical Package for the Social Sciences  for statistical analysis. All included infants were divided into two groups for the characteristic analysis of ROP with late GA and large BW: Group 1  and Group 2 (others). The absolute frequency  and 15 Type 1 ROP  infants were identified. All Type 1 ROP infants were treated by laser photocoagulation.p = 0.43) and Type 1 ROP  in Group 1 were comparable with those in Group 2. Group 1 infants were less likely to be diagnosed as respiratory distress  and less likely to have received blood transfusion  or supplemental oxygen  than Group 2. The information on the mode of oxygen delivery among infants in two groups are shown in p < 0.001), blood transfusion , and oxygen administration  among ROP patients were revealed in Group 1 than those in Group 2. We compared Group 1 infants with and without any ROP .As shown in  any ROP . VaginalThe numbers of premature infants who exceeded the screening criteria and the ROP infants (any ROP and Type 1 ROP) who may be missed are shown in The incidence of any ROP in China has been well investigated, ranging from 9.4%  to 17.8%A low-cost ROP screening could potentially reduce the disease burden of the family and the healthcare system costs. However, screening all the premature infants with late GA and large BW is stressful and costly, how to balance the risk of missed diagnosis and the reduction of screening number of infants is clinically significant. In the current study, besides conventional risk factors of blood transfusion and respiratory distress , vaginalIn a 7-year study period before and after the policy adjustment of guidelines on supplemental oxygen delivery, the number of premature infants screened and the incidence of Type 1 ROP increased by years, with a downward trend of BW. These findings suggest that the quality of neonatal care has improved over time, and it may be attributed to the decreases of transcutaneous oxygen saturation and the volume fraction of oxygen, and the use of pulmonary surfactant .There are several limitations in this study. First, this is a single center study conducted in a neonatal department of one tertiary general hospital with a relatively small sample size, especially for Type 1 ROP. Second, the proportions of vaginal delivery were very low in both two groups, which suggested that these infants may have other health issues apart from prematurity. The included premature infants may be more representative for those with sicker health conditions in the neonatal department of tertiary hospitals. Therefore, the association between vaginal delivery and ROP revealed in this study should be interpreted with caution. In order to develop suitable ROP screening criteria in South China, future multicentre studies with a larger sample size in other facilities with different health care levels and located in other areas should be used for analyzing the characteristics of ROP patients, how rates of Type 1 ROP vary and change over time, and the characteristics of the infants affected. In addition, limited by the retrospective nature of the study, information on the socioeconomic condition of patient families among premature infants was unavailable. This important information will be investigated in our following prospective studies.The incidences of any ROP and Type 1 ROP among premature infants with late GA and large BW were comparable with those of infants within the screening criteria in China. Furthermore, the increased number of infants screened, increased proportion of Type 1 ROP, and decreased BW of Type 1 ROP during the 7-year study period probably reflect improvements in neonatal care following the new revised policies in 2013, which have led to greater survival of extremely premature infants, but more mature infants are still being exposed to risk factors for ROP. Findings from this study suggest that screening criteria in China should be expanded to include more mature infants. ROP screening is recommended to those delivered by vagina among the premature infants with late GA and large BW.The original contributions presented in the study are included in the article/The studies involving human participants were reviewed and approved by the Institutional Review Board of Sun Yat-sen Memorial Hospital. Written informed consent for participation was not provided by the participants' legal guardians/next of kin because: all infants were anonymized and de-identified before being analyzed and this study was exempted from participant consent by the Institutional Review Board of Sun Yat-sen Memorial Hospital.DL, YLa, XG, and YLi contributed to the conception or design of the work. YLi, XG, LW, ZL, and YZ contributed to the acquisition of data. DL, LW, and DY contributed to the analysis. DL, XG, and YLi contributed to the interpretation of data for the work. DL and XG contributed to drafting the work and revising it critically for important intellectual content. YLa agree to be accountable for all aspects of the work. All authors gave final approval of the version to be published.This study was funded by the National Natural Science Foundation of China (82000946), Natural Science Foundation of Guangdong Province (2021A1515012238), and Science and Technology Planning Project of Guangzhou (DL). The funders played no role in the study design, data collection and analysis, decision to publish, or preparation of the 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.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.


Proteus spp., enteropathogenic Escherichia coli) decreased, and the content of some beneficial bacteria  changed compared to the control group. Unlike the C1 group, in the Bf group there was no decrease in the specific activities of maltase and alkaline phosphatase in the mucosa of the upper intestine, but the specific activity of maltase was decreased in the colon chyme compared to the control and C1 groups. In the Bf group, the specific activity of aminopeptidase N was reduced in the duodenum mucosa and the colon chyme compared to the control group. We concluded that indigenous bifidobacteria can protect the microbiota and intestinal digestive enzymes in the intestine from disorders caused by dysbiosis; however, there may be impaired motor function of the colon.In recent years, great interest has arisen in the use of autoprobiotics (indigenous bacteria isolated from the organism and introduced into the same organism after growing). This study aimed to evaluate the effects of indigenous bifidobacteria on intestinal microbiota and digestive enzymes in a rat model of antibiotic-associated dysbiosis. Our results showed that indigenous bifidobacteria (the Bf group) accelerate the disappearance of dyspeptic symptoms in rats and prevent an increase in chyme mass in the upper intestine compared to the group without autoprobiotics (the C1 group), but significantly increase the mass of chyme in the colon compared to the C1 group and the control group . In the Bf group in the gut microbiota, the content of opportunistic bacteria ( Human and animal health largely depends on the interaction between the intestinal epithelium and its microbiota. The main role in the implementation of this interaction belongs to the large intestine, which contains the largest number of microorganisms involved in the breakdown of undigested food and endogenous components  ,2,3,4. ABacteria belonging to the genus Bifidobacterium are among the main constituents of the intestinal microbiota in humans and animals ,6,7,8,9.At the same time, bifidobacteria participate in the modulation of the immune system   and restIn recent years, autoprobiotics—indigenous bacteria grown in laboratory conditions—have attracted great interest, as means for the correction of intestinal dysbiosis . This apDespite significant advances achieved in recent years into these topics, the action of individual probiotics and, especially, autoprobiotics in relation to the correction of the microbiota and the restoration of the digestive and protective functions of the intestine in most cases has not yet been sufficiently studied. Knowledge of these issues is especially important when using auto-probiotics for personalized therapy.The purpose of this work was to study the effects of autoprobiotic bifidobacteria on microbiota and intestinal membrane enzymes involved in digestion and maintaining homeostasis using a rat model of antibiotic-associated dysbiosis.Wistar rats , obtained from the Animal Breeding Center,  were kept in separate cages under constant conditions: Room temperature (18–22 °C), 12-h light/dark cycle, a noise level of no more than 85 dB, humidity of 50–60%. They were given free access to water and standard feed . The study carried out in strict accordance with the necessary ethical requirements and in accordance with the principles of humane treatment of animals (European Community No. 86/609 EC). The study was approved by the local ethics committee of the Institute of Experimental Medicine.®  at a dose of 75 mg/kg and metronidazole ®  at a dose of 50 mg/kg for three days [Experimental intestinal dysbiosis was induced in rats via daily intragastrical administration  of ampicillin ree days ,13,14.Bifidobacterium spp. were isolated from the Blauroca broth  after incubation of fecal samples for 48 h at 37 °C anaerobically. Five typical colonies grown at the bottom of the tubes were subcultured onto fresh culture medium to obtain a pure culture. Individual clones of bifidiobacteria were identified using microscopy and employing a RT-PCR set of primers encoding for 16S rRNAsequence  at a concentration of 5.5 × 108 CFU/mL and stored at −20 °C until use. PBS composition: 8.00 g/L NaCl, 0.20 g/L KCl, 1.44 g/L Na2HPO4, 0.24 g/L KH2PO4 (pH 7.4).For the preparation of auto-probiotic bifidobacteria, the feces were taken from healthy rats seven days before the administration of antibiotics. Individual clones of sequence . They weIn this study, we did not intend to identify specific species of bifidobacteria.The rats were divided into three groups : Control group C0 , group C1, and group Bf. The rats in the groups Bf and C1, after application of antibiotics for 3 days, received then for 5 days autoprobiotic bifidobacteria and PBS (instead of auto-probiotic), respectively. Rats in the control group C0 received water instead of antibiotics and PBS instead of auto-probiotic at the same time .It is important to note that the rats of the Bf group throughout the entire period of the experiment (including the period with the introduction of autoprobiotic bifidobacteria) were kept in separate cells in order to avoid the exchange of microbiota with other animals of this group. In addition, the diet of rats in our experiments did not contain bifidobacteria. These facts confirm that in our experiments, the rats of group Bf received only indigenous bifidobacteria. Fecal samples harvested on days 0 and 9 of experiments from all animals were used for microbiota study by qPCR and by metagenome analysis.After the necropsy examination (ninth day of the experiments), the state of the intestine was assessed macroscopically. The small intestine, the large intestine, and the cecum were weighed separately. The samples of mucosa and chyme were taken from various parts of the small intestine  and from the colon. They were stored at −80 °C and frozen to determine the activity of digestive enzymes of the intestinal epithelium and the chyme.The fecal samples were analyzed by 16S rRNA gene-based metagenomics analysis. Changes in the gut microbiota content were investigated by performing 16S rRNA gene-based metagenome analysis using a previously described approach . ChangesqPCR system for microbiota analysis  was used to characterize the main gut bacterial groups by quantitative  PCR. PCR amplification and detection were performed with Mini Opticon .The necessary temperature regime of the reaction was established . The probes’ DNA were labeled by HEX or FAM dyes. Analysis of PCR results in real time was carried out on the Bio-Rad device. Quantitative evaluation of the analyzed bacteria in the sample was noted on the starting point of the exponential growth curve and the program determined the cycle of amplification that corresponded to the point (threshold cycle C (t)). A special table (test system “Colonoflor”) was used to determine the number of CFU of bacteria contained in a test sample of feces. It could be used to correlate the value of the obtained threshold cycle C (t) with the number of CFU of bacteria in the sample.DNA from feces for 16S rRNA gene-based metagenome analysis was isolated using the QIAamp DNA Stool Mini Kit  following the manufacturer’s protocol. Samples were incubated in the lysis buffer at 90 °C for 10 min for optimal bacterial lysis. For microbiome sequencing, DNA libraries were prepared using the Illumina Nextera Sample Preparation Kit with DNA primers corresponding to V3–V4 regions of the 16S rRNA. Illumina MiSeq was used for sequencing the libraries .Simultaneous clustering of reads of all analyzed samples yielded 7782 OTUs. Most of the OTUs were present in both a low number of samples and low absolute amounts, which is a common feature of gut microbiome samples. Most of these small groups of reads failed to match any sequences from database Greengene 13.5 used for annotation. The number of annotated OTUs was 534 with the percent of unclassified reads being −22.4%. OTUs present in less than 5% of samples were discarded for noise filtering.http://www.bioinformatics.babraham.ac.uk/projects/fastqc (accessed on 20 November 2020)) was used to evaluate the quality of raw reads. CD-HIT-OTU-Miseq was used for OTU retrieval. CD-HIT-OTU-Miseq allowed us to retrieve OTU from paired-end reads without merging paired sequences. This was achieved by matching the clustering results for R1 and R2 reads. CD-HITOTU-Miseq could use only high-quality regions of reads for clustering. Clustering was performed using the following parameters: Lengths of high-quality regions of R1 and R2 reads of 200 and 180 bp, respectively, 97% read similarity for clustering cutoff and 0.00001 for abundance cutoff. OTUs were annotated using Greengenes database version 13.5 [Fastqc , alkaline phosphatase , and aminopeptidase N  activities were determined in homogenates of chyme from different segments of the intestine using the methods described earlier ,16. Chymt-test, Kruskal–Wallis test, ANOVA with post-hoc HSD test for unequal n, and MANOVA analyses in Python stats model; p < 0.05 was considered as significant.Statistical analysis was performed using the software package Statistica 8.0. . Differences between the groups were analyzed using Student’s When antibiotics were administered for 3 days in groups C1 and Bf, dyspepsia symptoms (which were expressed by the soft consistency of feces) was observed in animals. On the third day after the start of antibiotic administration, the number of animals with the soft consistency of feces was as follows: In the C1 group—4/15, and in the Bf group—3/10. In absence of autoprobiotics, after the withdrawal of antibiotics (the C1 group), the symptoms of dyspepsia in rats disappeared on the eighth day of the experiments, whereas in the presence of indigenous bifidobacteria (the Bf group), these symptoms disappeared already on the sixth day of the experiments.p < 0.05) . This increase in body weight in groups Bf and C1 did not differ from the weight gain in the control group C0 for the same period.On the fourth day after the start of antibiotic administration, the body weight of rats in the C1 group did not change, and in the Bf group it slightly decreased as compared to the first day. However, in the control group C0, the body weight of rats increased significantly from the first to the fourth days ( < 0.05) A. Subseqp < 0.05). On the fifth and on the seventh days of the experiments (the period of the introduction of the autoprobiotic or in its absence), the food consumption by the rats in the Bf and C1 groups increased in comparison with the first and third days of the experiments , as well as in comparison with the control group C0 on the seventh day of experiments (for each of the groups Bf and C1: p < 0.003).Food intake by rats in the control group C0 tended to decrease from 1 to 7 days of the experiments B. In grop < 0.05) compared to the C0 control group  in comparison with group C1, and did not differ from the corresponding values in the C0 group. It is noteworthy that in group Bf, the mass of chyme in the colon was significantly increased compared with groups C0 and C1   B.At the end of the experiments, an increase in the ratio of the weight of the cecum (including its contents) to the body weight of animal was also found in rats in the groups C1 and Bf compared to the control C0 .Bifidobacterium spp. in fecal samples from rats in group Bf was higher than in the control C0 and group C1 (p < 0.05). At the same time, the content of Proteus spp. and enteropathogenic Escherichia coli in the fecal microbiota in group Bf was reduced compared to group C1 (p < 0.05) and did not differ significantly from control C0   in group Bf, their relative abundance was noticeably higher than in group C1 (p < 0.05 or p < 0.01).The analysis of the metagenome revealed that in group Bf, the relative abundance of bacteria belonging to the genera  < 0.01) A–G. In g < 0.01) A–C. At tWhen examining alpha diversity, no significant differences were found between groups C0, C1, and Bf. However, it should be noted that biodiversity was highest in group C0 and lowest in group C1 .Bacteroides, Bifidobacterium, Blautia, Dorea, Lactobacillus, Oscillospira, Prevotella, Ruminococcus) in different groups using subsequent MANOVA analysis in a Python statistical model. Statistically significant differences were revealed between groups C1 and C0  of sample OTU compositions is presented in Bacteroides, Bifidobacterium, Blautia, Dorea, Lactobacillus, Oscillospira, Prevotella, Ruminococcus) and the correlations between them, we used an approach when all three groups were analyzed together. However, the correlations between the representation of the individual genera of bacteria was not established , only a tendency toward a decrease in the specific activity of maltase in mucosa of the upper intestine  was observed compared to the control group C0 A. With tp < 0.05) in the colon compared with the control group C0 (p < 0.05), and it was close to the C0 control group in the colon.The specific activity of aminopeptidase N (APN) in the mucosa in group C1 (without autoprobiotics) changed insignificantly in the duodenum and jejunum, but was markedly increased . Differences in this indicator between groups C1 and Bf were also noticeable (p < 0.01).Data presented on p < 0.01) and was characterized by a tendency to increase in comparison with the control group C0 (p < 0.05)  C.It is generally accepted that probiotics, which most commonly include strains of lactobacilli and bifidobacteria, can improve intestinal microecology and, as a result, also have a positive effect on many metabolic and physiological processes in the body, in the regulation of which they are involved ,6,7,8,10In this work, we used a rat model of antibiotic-associated dysbiosis in order to evaluate the probiotic efficacy of indigenous bifidobacteria on microbiota and membrane enzymes involved in digestion and maintaining intestinal homeostasis.Blautia and the family Prevotella in the chyme of this region .Our results showed that the use of indigenous bifidobacteria (autoprobiotic) in rats after the withdrawal of antibiotics (the group Bf) accelerated the disappearance of dyspeptic symptoms of intestinal dysbiosis in them, did not change the weight gain of animals, and prevented an increase in chyme mass in the upper intestine, which occurred in the absence of autoprobiotics (the group C1). However, there were also undesirable consequences. In the Bf group and the C1 group, there was a decrease in the mass of the mucosa in the colon and an increase in the mass of the cecum including its contents compared with the control C0. But a more significant undesirable consequence was in the Bf group: The mass of the chyme in the colon significantly increased compared with the control group C0 and the C1 group. This effect may be due to the indirect action of indigenous bifidobacteria (by changing the content of some other bacteria or their metabolites) on the motor function of the colon. This assumption was supported, for example, by the fact that we found that in rats of the Bf group there was a correlation between the mass of the chyme in the colon and the content of bacteria of the genus Proteus spp. and enteropathogenic Escherichia coli in the Bf group compared to the group C1. Thus, these data confirm the effectiveness of indigenous bifidobacteria in suppressing opportunistic bacteria. Moreover, the Bf group showed an increased content of Bifidobacterium spp. compared to control group C0 and group C1. The latter fact was in good agreement with our data obtained by metagenomic analysis. In addition, metagenomic analysis revealed changes in the relative abundance of a number of beneficial bacteria in the group Bf compared to the control group C0 and the group C1. Thus, in group Bf, the relative abundance of bacteria in the genera Ruminococcus, Prevotella, and Oscillospira was reduced and in the genera Bifidobacterium spp., Blautia spp., and Dorea spp. was increased in comparison with the control group C0. The physiological significance of these changes on body systems remains unclear.The results of the study of the microbiota in the feces of rats by qPCR showed a reduced content of opportunistic bacteria: We also identified specific effects of indigenous bifidobacteria on the activities of three important intestinal enzymes: Maltase, alkaline phosphatase, and aminopeptidase N.Maltase plays a key role in the breakdown of carbohydrates in adult mammals, participating in the final stages of hydrolysis of dietary glucose polymers, mainly starches . In our Regarding the responses of the specific alkaline phosphatase activity in the intestinal mucosa to the introduction of an autoprobiotic or its absence after the withdrawal of antibiotics, in our experiments, basically the same regularities were observed as in the case of maltase. This is especially important due to the fact that APh, in addition to participating in the digestion of phosphoric acid esters and regulation of lipid absorption ,17,18, aAPN, being a key enzyme in the final stages of hydrolysis of food proteins, is also involved in the degradation of biologically active peptides , in cholIn conclusion, the present study, using a rat dysbiosis model, showed that indigenous bifidobacteria can protect the microbiota and membrane enzymes, involved in digestion and maintaining homeostasis in the intestine, from disorders caused by dysbiosis; however, there are features in changing the listed parameters and an undesirable consequence in relation to the motor function of the colon which must be taken into account in the case of using such an autoprobiotic.


Chernozem, Luvisol, Planosol, and Cambisol. The soil samples were assayed for the content of total organic carbon (TOC) and the particle size distribution. HAs were extracted with the Schnitzer method and analysed for the elemental composition, spectrometric parameters in the UV-VIS range, and hydrophilic and hydrophobic properties, and the infrared spectra were produced. The research results have shown that the properties of HAs can be modified by the land use and the scope and that the direction of changes depends on the soil type. The HAs of Chernozem and Luvisol in the agri-ecosystem were identified with a higher “degree of maturity”, as reflected by atomic ratios , absorbance coefficients, and the FT-IR spectra, as compared with the HAs of the meadow and forest ecosystem. However, as for the HAs of Cambisol, a higher “degree of maturity” was demonstrated for the meadow ecosystem, as compared with the HAs of the agri- and forest ecosystem. The present research has clearly identified that the content of clay is the factor determining the HAs properties. Soils with a higher content of the clay fraction contain HAs with a higher “degree of maturity”.Many studies report organic carbon stabilization by clay minerals, but the effects of land use and soil type on the properties of humic acids (HAs) are missing. The aim of the paper is to determine the effects of land use and soil types on the characteristics of HAs, which have a considerable influence on organic matter quality. It was hypothesised that the effect of the land use on HAs properties depends on the particular size distribution. The research was performed in three ecosystems: agricultural, forest, and meadow, located in Slovakia. From each of them, the samples of 4 soil types were taken: The online version contains supplementary material available at 10.1007/s11356-021-14616-9. The role of organic matter (OM), including humic substances (HS), in the soil formation and development is unquestionable. OM is also an important indicator of soil fertility, and it plays an essential role in the efficiency of agriculture, the quality of the environment and global climate changes  changes  reservoirs is the soils of grasslands and forest soils. For that reason, the meadow and forest ecosystems are a land use preventing OC losses in soils. The OC losses triggered by soil processes are gas emissions of COMany reports point to progressive decreases in HS concentrations in soils converted from forest to arable land  has also been a technique used for the structural characterization of HS, especially for the identification of functional groups within the humus macromolecule Tan .Some essential information on the transformation of HAs is also provided by the results of high-performance liquid chromatography. Woelki et al. , Preuße The aim of the paper has been to present the land use and soil types effect on the characteristics of HAs determining the OM quality in the soil of various types. There are many results on the processes of OC stabilization by binding clay; however, some results on the changes in elemental composition and the other properties of HAs depending on the land use and soil types are missing. It was hypothesised that the effect of the land use on the properties of HAs will depend on the particle size distribution (soil types). The research has been performed in three ecosystems: agricultural, forest, and meadow, located in various parts of Slovakia. The properties of HAs have been determined from the assay of the elemental composition, spectroscopy in the UV-VIS, and IR ranges and the HIL-HOB properties applying the high-performance liquid chromatography.Chernozem (Ch), Luvisol (Lu), Planosol (Pl), and Cambisol (Ca) (IUSS Working Group WRB The experiment included 4 soil types: Chernozem (Ch) comes from the location of Voderady , found on the northern border of the Danube Basin. The geological structure shows the Neogene strata, which consist mainly of claystones, sandstones, and andesites, covered with younger quaternary rocks represented by different fluvial and Aeolian sediments  comes from the location of Vavrecka , found in Horna Orava, which is part of the Flysch Belt. The geological structure shows the alternation of clay slates, sandstones, and conglomerates in the layers of various depth. It is located in a cold climate region with an average annual temperature of 4.6°C and the total annual precipitation of 1010 mm  comes from the location of Prietrz , on the Myjava Hills. The geological structure includes the Carpathian flysch, which consists mainly of marine deposits of claystones, shales, and sandstones  comes from the location of Plave Vozokany , in the northeast of the Danubian Upland. The geological structure involves clay, gravels, and sands covered with quaternary sediments . The distance between the replications was 20 m in equilateral triangle. The distance between fields differed, depending on the locations; however, it was not more than 2 km. The distance between the locations   with 0.05 M HCl (1:10 w/v). After centrifugation the residue was rinsed with distilled water till neutral.The extraction (24 h) of the remaining solid with 0.5 M NaOH (1:10 w/v), with occasional mixing, followed by centrifugation.The precipitation (24 h) of HAs from the resulting alkaline extract with 2 M HCl to pH=2 and centrifugation.2O, 5 mL HCl, 5 mL HF) over a 24-h period, followed by centrifugation. This procedure was repeated three times. The HAs residue was treated with distilled water until a zero reaction to chloride was achieved.The purification of the resulting HAs; the HAs residue was treated with a mixture of HCl/HF . The H/C, O/C, O/H, and N/C atomic ratios and ω  were calculated using the following formula:The HAs separated were analysed for:280), 400 nm (A400), 465 nm (A465), 600 nm (A600), and 665 nm (A665) and used to calculate the coefficient values:UV-VIS absorption spectra . VIS spectra were obtained from 0.02% HAs solutions in 0.1 M NaOH and UV-spectra after fivefold dilution. The absorbance was measured at 280 nm (A2/4 – 280 nm and 465 nm absorbance ratioA2/6 – 280 nm and 665 nm absorbance ratioA4/6 – 465 nm and 665 nm absorbance ratioA400 - log A600 (sKumada 1987);ΔlogK = log A−1 were recorded for HAs (3 mg) in KBr (800 mg). To increase the legibility of the spectra, deconvolution was applied, with a filter making the bands of γ = 4 narrower and using the process of smoothing, for which the length parameter was l = 80%  over 400–4400 cm−1; the injection of the sample was 10 μL; solvent – acetonitrile – water; solvents flow in the gradient (ratio H2O:ACN (v/v) over 0–6 min – 99.5: 0.5, 7–13 min – 70: 30, 13–20 min – 10: 90); detection – at the excitation/emission wavelength (λex/λem) of 270/330 nm. Based on the areas determined under peaks, the share of HIL and HOB (∑HOB = HOB-1 + HOB-2 + HOB-3) fractions in HAs molecules and parameter HIL/∑HOB was determined , forest (FE), and meadow ecosystems (ME)) within a soil type was evaluated with Duncan’s post hoc test at p < 0.05.280, A465, A665, A2/4, A2/6, A4/6, and ΔlogK), as well as the HIL-HOB properties. The method involves dividing the data set into groups to produce clusters where the elements are similar to one another and, at the same time, different from the elements of the other groups. The relationships between the share of clay, silt, and sand fraction and the basic parameters of HAs were defined using the Pearson’s correlation coefficients (P ≤ 0.05). The above relationships were determined using statistics software STATISTICA MS 12.The effect of the soil type and the land use on the properties of HAs was defined with cluster analysis. The method involves dividing the data set into groups to produce clusters in which the elements are similar to one another and, at the same time, different from the elements from the other groups. The groups of similar treatments are presented in a form of dendrogram. In a given group, the smaller the Euclidean distance, the more similar the objects. Data clustering was performed with the Ward method. The analysis was made after data standardization. The cluster analysis was performed based on the elemental composition , spectrometric parameters . Cambisol demonstrated a similar content of sand and clay fraction  and Planosol a similar content of silt and clay. The lowest content of TOC was reported for Cambisol and the highest for Planosol. The analysis of correlation did not show a significant dependence between the content of clay and the content of TOC; however, as reported in literature, one cannot exclude the effect of clay on the stability of SOM . The soils differed significantly in terms of the particle size distribution Table . The higLuvisol sampled from the forest ecosystem. The lowest share of that element as well as N was found for the HAs of Planosol (FE). The lowest content of H and the highest content of O were recorded in the HAs of Chernozem from the agri-ecosystem (AE).According to De Moraes et al. , the chaTable Chernozem in the agri-ecosystem, while the lowest values of the O/H ratio and the highest of the N/C ratio were found for the HAs of Chernozem of the forest ecosystem. The above dependencies point to the HAs of agri-ecosystem showing the highest and the HAs of the forest ecosystem—the lowest “degree of maturity” . Generally, the HAs of Luvisol showed the highest contents of C and lower contents of H, as compared with the HAs of the other soils  to 29.46% (Ch FE)  Table . With thE) Table . IrrespeE) Table  were ideChernozem and Luvisol under agricultural use showed a higher band intensity in the range of 1730–1710 cm−1, as compared with the molecules of the HAs of the forest and meadow ecosystem. The intensity of the bands in the range of 3400–3100 cm−1 and 2960–2920 cm−1 was highest for the HAs of the forest soils. The HAs of Luvisol, Planosol, and Cambisol of the forest ecosystem demonstrated the lowest intensity of the bands in the range of 1500 to 1000 cm−1. As for the HAs of Chernozem, the lowest band intensity for the range from 1660 to 1000 cm−1 was recorded for the soils under agricultural use.Figure Planosol and Cambisol as well as the HAs of the forest ecosystem of Cambisol and HAs Planosol under agricultural use. In the second subgroup, the most similar properties were found for the molecules of the HAs of Chernozem in the forest and meadow ecosystem. In the second group, similar properties were noted for the HAs isolated from Luvisol, irrespective of the ecosystem, and the HAs extracted from Chernozem sampled from the agri-ecosystem. As seen from the diagram, both the soil type and the land use are the factor which determines the properties of HAs. With some approximation, one can assume that Luvisol is the type of the soil which is modified even least considerably by the land use.To acquire complete information on the differences (similarities) in the chemical composition of HAs, depending on the soil type and the land use, the cluster analysis was applied based on the elemental composition, spectrometric parameters, as well as the HIL-HOB properties dividing the HAs into two groups Fig. . In the 2/4, A2/6, A4/6 and positive between the content of C and parameter ω and the values of absorbance) are characteristic for the soils with a higher clay fraction content and a lower sand fraction content. Even earlier Jindaluang et al.  values of the coefficient of correlation between the clay fraction content and the content of C in the molecules of HAs, the values of the degree of internal oxidation and absorbance values, and the negative (significant) correlations between the content of the clay fraction and the content of H and the values of the H/C, AESM 1(DOC 61 kb)