Circulating protein levels were measured by ELISA. RNA was extracted from AT and expression levels were relatively quantified by PCR. Results: After 1 year, no significant difference in the change in the investigated markers between the intervention group and the control group was observed. Changes in circulating adiponectin and VO2 peak correlated in the total population . At baseline, circulating adiponectin and TNF correlated inversely with insulin and with C-peptide and VO2peak, respectively . Conclusion: In this population with concomitant diabetes and CAD, ATs’ inflammatory profile remained unchanged apparently after 1 year of exercise intervention. Changes in the VO2peak were nevertheless, related to changes in circulating adiponectin levels. Trial registration: http://www.clinicaltrials.gov NCT01232608.Introduction: Adipokines, expressed by adipose tissue (AT), have been associated with metabolic disturbances and coronary artery disease (CAD). The impact of exercise training on the AT in patients suffering from both diabetes and CAD is unknown. To gain knowledge on changes in ATs’ inflammatory profile in such a population, we investigated the effects of long-term exercise on selected adipokines and their associations with physical performance and glucometabolic variables. Adiponectin was selected based on its anti-atherogenic and anti-diabetic properties and visfatin and tumour necrosis factor (TNF) for their association with atherosclerosis and metabolic disorders. Not many studies have focused on the effects of long-term exercise training on adipokines in patients with concomitant T2DM and CAD. Methods: Patients with type 2 diabetes and CAD (n = 137), 41–81 years, 17.2% females, were randomized in a 1:1 manner to an exercise group, who underwent 1 year of 150 min weekly combined strength and endurance exercise, or a control group. AT from the gluteal region and blood samples were obtained at baseline and after 12 months, along with a physical performance test, assessed by the VO Adiponectin is an adipocyte-derived protein that plays a major role in metabolic disorders such as type 2 diabetes (T2DM), insulin sensitivity, metabolic syndrome (MetS) and coronary heart disease. Although mainly produced by the adipose tissue (AT), several reports have shown that scarce amounts of adiponectin are produced by different tissues and cells, such as cardiomyocytes, skeletal muscle, osteoblasts and the liver. With itAltered properties of the AT with a subsequent reduction in adiponectin and increased TNF secretion may thus result in an imbalance of the glucose homoeostasis, precipitating metabolic disorders. Circulating TNF, shown to be increased in CAD patients, seems tContrary to TNF, visfatin seems to bind and sensitize the insulin receptor, reducing glucose release from hepatic cells and increasing glucose consumption in adipocytes, thereby causing decreased circulating glucose levels. AT exprTNF has been reported to be strongly involved in the regulation of both adiponectin and visfatin, as shown by the interesting study of Hector et.al.Physical activity is effective in primary prevention of CVD, partly by regulating glucose metabolism and increasing the action of insulin ,24. Alth2 peak were further explored.To obtain more insight into the connection between the serum levels and AT-expressed adipokines with metabolic traits, exercise and physical fitness, we investigated effects of one-year exercise training on adiponectin, visfatin and TNF in patients with concomitant CAD and T2DM. Our hypThe present investigation is a sub-study of the EXCADI (exercise training in patients with coronary artery disease and type 2 diabetes) trial, in which patients with T2DM and CAD were included at Department of Cardiology, Oslo University Hospital – Ullevaal, between August 2010 and March 2012. The study design has been described previously. In brie2 peak was defined as the highest consecutive 30s average of oxygen uptake during the test, measured by breathing through a two-way breathing mask. In the main EXCADI study, we could not demonstrate effects of exercise intervention on the VO2 peak or HbA1c. However, these parameters did improve in the subgroup of patients without advanced vascular disease[A cardiopulmonary exercise test (CPET) was performed at the beginning and at the end of the study. A VOr disease.The EXCADI study was conducted in accordance with the Declaration of Helsinki. It was approved by the Hospitals Data Protection Services and the Regional Committee of Medical Research Ethics in South-Eastern Norway (2010/1060) and all patients gave their written informed consent to participate.The physical exercise program was designed in collaboration with the Norwegian School of Sport Sciences. The details of the program have been described previously. BrieflyVenous blood samples were collected by standard venipuncture between 08.00 am and 10.00 am in fasting condition, before intake of morning medication both at inclusion and after 12 months. AT samples were taken from the gluteal region and snap frozen immediately at −80°C until extraction of RNA. Blood samples, including HbA1c, insulin and C-peptide, were acquired and measured through conventional routine methods. Serum was prepared for establishing a biobank by centrifugation within 1 hour for 10 min at 2500 x g. Determination of circulating adiponectin, visfatin and TNF was performed by the following ELISA methods: Human total Adiponectin/Acrp30  and Human TNF-α  (both R&D Systems and Human Visfatin)  (MyBioSource). The inter-assay coefficients of variation in our laboratory were 8.0%, 4.2% and 8.1%, respectively.Total RNA from AT was isolated, including disruption and homogenization in Tissue lyser (Qiagen), by the use of a High Pure RNA Tissue Kit  (Hoffman-La Roche Ltd.), according to a combination of the kit protocol and previous experience in our laboratory. Minor modification included adding Proteinase K after the homogenization procedure for the inactivation of DNAses and RNAses. RNA quality and quantity (ng/µL) were determined using a NanoDrop™ 1000 Spectrophotometer (Nanodrop Technologies). Extracted RNA was stored at −80°C until analyses. Copy DNA (cDNA) was synthesized from an equal amount of RNA (5 ng/ųl) with qScript™ cDNA superMix  (Quanta Biosciences Inc.). Real-time PCR was performed with TaqMan Low Density Custom Arrays on a ViiA™7 instrument, using TaqMan® Universal PCR Maser Mix (P/N 4324018) and TaqMan® assays for adiponectin (Hs00605917_m1), visfatin (Hs00237184_m1) and TNF (Hs01113624_g1) . β-2-microglobulin (Hs99999907_m1) (Applied Biosystems) was used as the endogenous control, and mRNA levels were determined by relative quantification (RQ) using the ΔΔCT method.th and 75th percentile) for skewed data. The Mann-Whitney U test or Student’s t-test was used for analysis of continuous variables and Chi-square test for categorical variables as appropriate for differences between groups. The Wilcoxon Signed Rank test was used for analysis of within-group changes after intervention. Differences in changes during the intervention period (delta) between the randomized groups were analysed by the Mann- Whitney U test. Correlations analyses were analysed by Spearman’s rho and adjusted for by Bonferroni correction. Power calculations were performed in the main study based on expected HbA1c reductions, thus not for considering effects on the investigated adipokines. Statistical calculations were performed using SPSS version 26 . P-values <0.05 were defined as statistically significant.The demographic data are given as proportions, mean (±standard deviation) for data with equal distribution or as median . AT expression of adiponectin was inversely correlated with BMI and positively correlated with the VO2 peak . Circulating adiponectin correlated inversely with insulin, C-peptide and HOMA2-IR , but not with BMI or VO2 peak. Circulating TNF also correlated with insulin, C-peptide and HOMA2-IR , with an additional inverse correlation with the VO2 peak . After Bonferroni correction (48 comparisons), the correlations between circulating adiponectin and insulin (r = −0.271) and between circulating TNF levels and C-peptide and VO2 peak remained statistically significant .Associations between serum levels and gene expression of adipokines at baseline and after interventionWe investigated the correlations between serum levels and gene expression of the investigated adipokines at baseline and after 12 months. We found an inverse correlation between circulating visfatin and gene expression of TNF in the control group  and between circulating visfatin and visfatin gene expression in the exercise group  (Supplementary Table 1) at baseline. We also found an inverse correlation between circulating and genetically expressed TNF in the exercise group, after intervention  (Supplementary Table 2). The observed associations were no longer statistically significant after Bonferroni correction (18 comparisons).2 peakAssociations between changes in adipokines and the VO2 peak after 1 year, we observed a significant correlation between delta circulating adiponectin levels and the delta VO2 peak in the compliant population (n = 112)  as shown in When investigating delta values (i.e. changes from baseline to 12 months) for adiponectin, visfatin and TNF and VO2 peak and increased serum adiponectin levels.The main finding in our study was that long-term exercise intervention did not affect AT expression and circulating levels of adiponectin, visfatin and TNF significantly. We could, however, show an association between the improved VOPrevious studies examining the exercise effect on adiponectin suggest that a prerequisite to change in adiponectin levels, both genetically expressed in AT and circulating levels, is a 10–15% weight loss. In our A study performed on abdominally obese men showed an elevation in circulating adiponectin levels immediately after an acute bout of exercise compared to baseline, which remained elevated for at least 30 min of rest. One wouTNF increased significantly in the exercise group and non-significantly in the control group, so the difference in change was not significant. This could possibly be because in patients with longstanding CAD and T2DM, the AT may develop into a more pro-inflammatory state, potentially rendering rigorous exercise training in such patients non-beneficial. Although previous findings mainly suggest otherwise , the pop2 peak. This is consistent with previous findings[2 peak were shown to be independently related to low levels of circulating adiponectin[2peak[2 peak and circulating adiponectin, which, however, attenuated when adjusted for BMI. This suggests that BMI may be a confounder in these associations, which may further imply that the quantity of AT is the decisive factor for circulating levels of adiponectin[2 peak and the change in circulating adiponectin, which is in line with the aforementioned findings.When exploring baseline associations, we found that AT expression of adiponectin was inversely correlated with BMI and positively correlated with the VO findings. It is d findings. In a stiponectin. This istin[2peak. Animal tin[2peak. A studyiponectin. In our At baseline, we found an inverse correlation between circulating adiponectin and insulin, C-peptide and HOMA2-IR. However, after Bonferroni correction, only the inverse correlation between insulin and adiponectin remained significant. Although previous reports have mainly focused on the relationship between insulin resistance/insulin sensitivity and adiponectin, the results are somewhat in accordance with these findings, suggesting a protective role of adiponectin against insulin resistance and MetS ,41. TherWhen investigating the associations between circulating levels and gene expression of adipokines, we found an inverse correlation between circulating and AT-expressed visfatin and between circulating visfatin and TNF gene expression at baseline. After the intervention, we found an inverse correlation between circulating and genetically expressed TNF in the exercise group. However, none remained significant after bonferroni correction. This could be indicative of a downregulation at the gene expression level with increased circulating visfatin and TNF. Nonetheless, one has to consider the limited amount of representative AT samples, particularly for TNF. Furthermore, other potential sources for adipokines, such as deeper layers of the AT, have to be considered.AT expression and circulating levels of visfatin and circulating TNF increased, surprisingly in the exercise group, but the differences in changes from controls were not significant. Considering their potential deleterious effect on glucometabolism and cardiovascular system, we anticipated a reduction in TNFα and visfatin after exercise training; however, the majority of previous studies, although performed in different populations, have shown that circulating TNFα remains unchanged following exercise . Ryan et2 peak at baseline, which is significant after Bonferroni correction. In line with this, one study has shown significant improvement in the VO2 peak 4 months after treatment with anti-TNFα therapy[Nevertheless, we found that TNFα correlated inversely with C-peptide and teh VOα therapy.2 peak in our study may be multifactorial. Our population with many years of T2DM had developed vascular complications and were heavily medicated on ACE inhibitors, statins, platelet inhibitors and anti-diabetic medication that might have affected the outcome, which maybe due to some degree of irreversible changes in the AT composition. Furthermore, previous reviews on the effects of exercise on adipokines such as visfatin suggest that reduction in central fat percentage and change in AT composition are a necessity to observe changes in circulating levels of adipokines[Levels of visfatin have been shown to decrease in obese, non-diabetic and diabetic subjects, following 12 weeks of both aerobic and resistance exercise training, and the reduction seems to be related to changes in both glucose and insulin, but not necessarily to changes in weight . The absdipokines.The main limitation of our study is that it was designed to study the effect of exercise training on HbA1c , thus noThe absence of weight reduction is unfortunate. As mentioned, several studies suggest that loss of weight (through both diet and exercise) seems to be the main driving factor towards desired effects on adipokines, which is plausible, considering that AT is the main source. Whether the amount of exercise in our population was sufficient could also be discussed. Furthermore, any physical activity in the control group was not accounted for, which might have influenced the results of our study.Adiponectin circulates in blood in many isoforms and the high-molecular weight adiponectin is thought to be most biologically active in the uptake of glucose and insulin sensitivity. We haveOur study population was heavily medicated with ACE inhibitors/A2-blockers, statins and antidiabetics. Some of these medications have anti-inflammatory properties, such as statins and metformin, which might have affected the circulating levels of adiponectin and visfatin and thus masked the potential beneficial effect of exercise training ,54.The strength of our study was the randomization principle applied with equal distribution between both groups. The number of participants in our study exceeds numerous studies that have addressed the same issue.2 peak, suggesting a potential association with cardiorespiratory fitness.In our combined T2DM and CAD population, exercise training without weight loss had a limited impact on AT expression and circulating levels of adipokines. Adiponectin was associated with the VOClick here for additional data file.


The most numerous class of lipids were ceramides (n = 61), while the free fatty acids were the most abundant ones (average concentrations of 10−5 mol/L). The main advantages of apocrine sweat microsampling include: (a) the non-invasiveness of the procedure and (b) the unique feature of apocrine sweat, reflecting metabolome and lipidome of the intracellular space and plasmatic membranes. The SLIDE application as a sampling technique of apocrine sweat brings a promising alternative, including various possibilities in modern clinical practice.We designed a concept of 3D-printed attachment with porous glass filter disks—SLIDE (Sweat sampLIng DevicE) for easy sampling of apocrine sweat. By applying advanced mass spectrometry coupled with the liquid chromatography technique, the complex lipid profiles were measured to evaluate the reproducibility and robustness of this novel approach. Moreover, our in-depth statistical evaluation of the data provided an insight into the potential use of apocrine sweat as a novel and diagnostically relevant biofluid for clinical analyses. Data transformation using probabilistic quotient normalization (PQN) significantly improved the analytical characteristics and overcame the ‘sample dilution issue’ of the sampling. The lipidomic content of apocrine sweat from healthy subjects was described in terms of identification and quantitation. A total of 240 lipids across 15 classes were identified. The lipid concentrations varied from 10 The entire sweat gland system represents 1.6–5 million apocrine and eccrine glands distributed throughout the body surface . The eccTo characterize sweat glands by their secretory mechanism, they can be divided into three main groups: holocrine, merocrine  and apocrine . (1) The+, Cl−, Ca2+, K+, Mg2+, PO43−, NH4+ and others, including ionic species, and (3) organic molecules—amino acids, hydroxy/keto acids, free fatty acids, saccharides, urea, creatinine, uric acid, vitamins, volatile compounds and intact/metabolized exogenous chemicals and pharmaceuticals . Li. Li25]. −5 mol/L compared to low abundant LPE, LPC, PEO and PCO with a median of 10−9 mol/L (−10 to 10−4 mol/L). Moreover, the numbers of detected lipids significantly differed across the classes from only a few in the PI class up to dozens in Cer and TG classes.Concentrations of lipids in biological samples usually differ by many orders of magnitude. Similarly, in sweat extract a large dispersion of the concentration levels between and within lipid classes can be observed. The most abundant FA, CE, and DG classes lie in the median of 10−9 mol/L . FinallyIf the concentrations of all lipids in each class are summed, a large representation of FA (83.8%) followed by DG (7.9%), CE (4.4%), TG (2.1%) can be observed a. Sphingn = 60 samples), correlation coefficients 0.26 and higher were calculated as statistically significant, where medium and strong correlations were higher than 0.5 and 0.7, respectively. Systematic trends can be observed as lipids in one class generally behave similarly compared to another lipid class . For example, ceramides show a negative correlation with most other lipid classes except free fatty acids to which they correlate positively, and triacylglycerols with both positive and negative correlation regions on the heatmap. A positive correlation among all glycerophospholipids can be observed except for the negatively correlated PEO and PS classes.To overview the relationships between lipid classes in sweat, Pearson’s linear correlation was performed and the results visualized in a heatmap . Based ot-test and fold-change. When Benjamini-Hochberg correction  was applied, none of any of the lipids was evaluated as statistically significant. Despite systematic trends of increased ceramides, diacylglycerols, and partly decreased selected glycerophospholipids  can be seen, the relative changes are lower than 15% for the majority of lipids.In addition to the basic description of the lipid composition of apocrine sweat, the differences of sampling between the left and right armpits were studied. The visualization of all lipids categorized by lipid classes was performed using Cytoscape software . FinallyIn the study presented here, we focused on characterizing apocrine sweat as a potentially clinically relevant biofluid, from multiple points of view. First, we have developed and described a simple, easy-to-use sampling technique, using a custom 3D-printed attachment and a commercially available porous glass filter disk. This concept not only offers an opportunity for other researchers and clinicians to adopt our technique for further studies, but it also brings a unifying aspect to apocrine sweat sampling, as the 3D blueprint can be easily shared and modified for further development. We have also described a standardized protocol with recommendations for consistent sampling. The usage of antiperspirants and other cosmetics with underarm application needs to be avoided before the sampling of apocrine sweat as it can lead to obstruction of apocrine sweat glands . Due to Lipidomic analysis is increasingly showing its potential in the field of clinical diagnostics as well as offering unique insights into the biochemistry and biophysics of biological fluids and tissues . ApocrinDescriptive statistical evaluation of the studied material is necessary for its use in clinical applications. As sweat had already been shown to be a highly variable material as regards water content and concentration of ions ,20,30 anThe general principle and usefulness of the lnPQN were additionally proven by multivariate statistical analysis, particularly in the hierarchical cluster analysis, where 9 out of 10 individual participants were clustered, while an effect of the different day or side of sampling was not seen. Further insight into biological variability was offered by the results from PCA, in which all individual samples clustered close together, but 3 participants  lay further away from all other samples. This outcome highlights one imperfection of our study, namely the uneven representation of ages in our small cohort. These results are also in compliance with larger cohort studies investigating correlations between plasma lipids, age, and sex of participants, where intra-/interclass trends in lipids showed a good correlation with age .n = 61) were the most represented, followed by TG (n = 37), SM (n = 23), PC (n = 20), DG (n = 17) and FA (n = 17). At first glance the large number of ceramides of structurally heterogeneous character  may be striking; however, they are found abundantly as constituents of the cutaneous lipidome, where their function (water permeability barrier of the skin) and their role in pathobiochemical processes  have been well documented [In terms of the lipid concentration in apocrine sweat, FA dominated, followed by glycerolipids (TG and DG) and then CE and phospholipids (SP and GP). Takemura et al.  discovercumented ,36,37.To understand the biochemistry and biological behavior of a particular material, it is helpful to focus on the trends within and between groups of studied biomolecules in a particular biofluid or tissue. By constructing a correlation map, we could easily trace the relationship trends between lipid classes. We could also observe that one lipid class generally behaves in a consistent manner compared to another class, which is commonly found in studies of plasma lipidome . Glycerop-value correction (Benjamini-Hochberg). Furthermore, since these differences for the majority of lipids were only up to +/−15%, this contribution was minor compared to the interindividual variability of individual lipid classes (which are mostly greater than 50%). On the other hand, these differences could be of significant importance for diseases affecting only one of an organ pair , or they could also reflect the physiological dominance of the right or left hand and/or lymph nodes [To fully describe the studied material, differences between the sides of sampling were also evaluated. Although systematic trends in lipid class levels were observed between the left and right sides of sampling, these changes were considered nonsignificant after the application of ph nodes . HoweverThe results presented above show the potential of apocrine sweat sampling and its analysis by advanced mass spectrometry coupled with liquid chromatography, as a promising noninvasive alternative to established methods in routine clinical analysis. Based on the results of our research and also from other studies, it is likely that new unsuspected applications for various microsampling techniques will be emerging for both diagnostic and therapeutic purposes . Given t® LIPIDOMIX® Mass Spec Standard mixture and ceramide (d18:1-d7/15:0) were purchased from Avanti Polar Lipids . Arachidonic acid-d8 was acquired from Cayman Chemical Company .Acetonitrile (ACN), isopropanol (IPA), water, and ammonium acetate  were purchased from Sigma-Aldrich . SPLASHThe attachment was constructed in 3D Builder . The external dimensions of the attachment were 41 mm in length and 14 mm in radius . A 1.75 Apocrine sweat was collected in gloves using a custom 3D-printed attachment with porous glass filter disk  and afterwards they were not allowed to use any cosmetic products with underarm application until after the sweat collection procedure. Washing with soap or body shampoo when showering or bathing the evening before the collection was allowed. The next morning, the axillary apocrine sweat was collected as follows: one hour prior to sampling, the armpits were cleaned with distilled water and lightly wiped with a paper towel to remove surface dirt, microbial debris, dead cells and sebum, then the filter disk was placed on the attachment and it was carefully immersed in 80% isopropanol (so that it just touched the surface). The axillary apocrine sweat was then removed by passing the disk over the surface of the skin of the shaved part of the axilla, where the apocrine sweat glands open into the hair follicles. Apocrine sweat was expressed by slight pressure on the disk and immediately drawn by capillary forces into the interior of the disk. The extraction was performed bilaterally, with an estimated extraction time of approximately half a minute per axilla. One disk was always used for sampling only one side (left or right axilla). After the collection, the disk was pushed out of the attachment with tweezers (from the other side of the attachment) and transferred to a premarked tube, which was then immediately frozen at −80 °C until further sample preparation.To test the reproducibility of this novel sampling approach, apocrine sweat was collected from 10 healthy individuals (A–J) in three consecutive sessions (1–3) from both the left (L) and right (R) sides providing six samples from each individual and a total of 60 samples , and finally pipetted to glass vials for LC/MS analysis. An aliquot of 15 μL was taken and pooled from each extracted sample to make the final quality control (QC) sample. To eliminate potential systematic errors, QC were analyzed every 5th sample (for instrument stability control). Both orders of sample preparation and analysis were strictly randomized (for batch layout see Sweat samples were extracted from the porous glass filter disk by one-step isopropanol (IPA) extraction in random order. First, 400 µL of 80% IPA was added to the tube containing the disk and the mixture was sonicated for 10 min and incubated for 1 h at room temperature on a shaker. The extracts (200 µL) were transferred to Eppendorf tubes (1.5 mL), centrifuged  and the system was controlled by Analyst software . A reversed-phase BEH C8  column  was used for the chromatographic separation. Mobile phase A consisted of ACN: H2O , the mobile phase B was IPA: ACN , and both contained 10 mM ammonium acetate. The flow rate was 0.35 mL/min and the column was tempered at 55 °C. The elution gradient was set as follows: commencement with 32% B for 1.5 min; linear increase to 85% B at 15.5 min; increase to 97% B at 15.6 min maintained for 2.4 min. The gradient then decreased to the initial composition of 32% B at 18.1 min and this ratio was maintained for 1.9 min for column equilibration.The method for targeted lipidomic analysis, using liquid chromatography coupled to mass spectrometry, was adopted from Xuan et al. . The LC The parameters of the ion source and gases of the mass spectrometer were set as follows: ion spray voltage +4500 V and −4500 V; curtain gas, 40 psi; ion source gases 1 and 2, 60 and 50 psi respectively; source temperature 400 °C. Scheduled MRM mode with a window of 2 min was applied for the data acquisition. Positive and negative ionization of compounds in one analysis was performed, using the polarity-switching feature of the used mass analyzer.® LIPIDOMIX® Mass Spec Standard mixture (with the syringe flow rate set at 3 µL/min) simultaneously during the analysis of QC samples was carried out for the evaluation of matrix effects. Non-scheduled MRM transitions corresponding to each deuterated standard in the mixture were measured to investigate ion suppression or ion enhancement across the time of analysis (8 in positive and 5 in negative mode and the dwell time was set to 30 ms) (The adduct and fragmentation parameters  as well o 30 ms) . To uncoo 30 ms) .Data from the lipidomic analysis were integrated by SCIEX OS software  and processed in the R program  using the Metabol package . Peak art-test, box plots, violin plots) methods. The p-value was calculated by parametric t-test. The fold-change value was expressed as log two of the difference of medians of the given parameter of comparison. The Cytoscape program  was used for global visualization of changes occurring in lipid profiles [p-value  and the depth of color is based on the fold-change (shades of red/blue represent an increase/decrease between two tested groups) or the % of relative variability.Data were statistically evaluated in GraphPad  and SIMCA software . Ln and lnPQN transformation, Pareto scaling, and mean centring were applied to the final dataset. Data were evaluated by both multivariate  and univariate (profiles . Each of® LIPIDOMIX® Mass Spec Standard mixture with the addition of two more deuterated standards: arachidonic acid (d8) and ceramide (d18:1d7/15:0). A 6-point calibration curve was plotted for each standard representing the whole lipid class and the corresponding equations were used for calculating concentrations  was prepared using isotopically labeled SPLASHtrations . Values bundance , while t


Drosophila melanogaster as a model organism for the conjugation of plasmids of various incompatibility groups in the gut. Enterobacteriaceae conjugation pairs were identified in vitro and used for oral inoculation of the Drosophila gut. Flies were enumerated for the donor, recipient, and transconjugant populations. Each donor-recipient pair was observed to persist in fly guts for the duration of the experiment. Gut concentrations of the donors and recipients were significantly different between male and female flies, with females generally demonstrating increased concentrations. Furthermore, host genetics significantly altered the concentrations of donors and recipients. However, transconjugant concentrations were not affected by host sex or genetics and were detected only in the IncPε and IncI1 plasmid groups. This study demonstrates Drosophila melanogaster as a model for gut-mediated plasmid transfer.Horizontal gene transfer (HGT) is a driving force of microbial evolution. The gut of animals acts as a potent reservoir for the lateral transfer of virulence, fitness, and antimicrobial resistance genes through plasmids. Reduced-complexity models for the examination of host-microbe interactions involved in plasmid transfer are greatly desired. Thus, this study identifies the use of IMPORTANCE Microbial evolution in the gut of animals due to horizontal gene transfer (HGT) is of significant interest for microbial evolution as well as within the context of human and animal health. Microbial populations evolve within the host, and factors from the bacteria and host interact to regulate this evolution. However, little is currently known about how host and bacterial factors regulate plasmid-mediated HGT in the gut. This study demonstrates the use of Drosophila and the roles of sexual dimorphism as well as plasmid incompatibility groups in HGT in the gut. The emergence of antibiotic-resistant bacteria (ARB) is a threat to agricultural and human health and safety –4. Antib–4–8–21–The conjugation process leading to the exchange of small to large plasmids or integrative conjugative elements (ICE) from one bacterial cell to another is prevalent in every environment where bacterial communities persist. The gut of animals is a common area of focus for studying microbial communities and has been indicated as a potential habitat for subsequent plasmid-mediated conjugation and HGT , 24–28. 24–Drosophila melanogaster is an arthropod used as a model for study in a multitude of scientific fields, from biopharmaceuticals to behavior and cellular development  act as laboratory models for other significant arthropod hosts such as mosquitos (Culicoidea) and houseflies (Musca). For many of these arthropod pests, a primary food source is the fecal waste of agricultural animals in natural and built environments. Similarly, the fruit fly diet consists of the microbes and nutrients found on the surfaces of decomposing fruit, including yeast, molds, as well as dietary and contaminating bacteria. These fruit waste and decomposition sources occur in complex agricultural environments that have been demonstrated to be potent reservoirs of antimicrobial resistance and pathogens   , pCVM291(MM0001) , or pC20(MM0001)  and the ectively . TranscoDrosophila of either the W1118 or CantonS fly strain for all three conjugation pairs tested . Likewise, the HS-4 recipient colonized the guts of male and female CantonS flies at 5.41 ± 2.61 and 2.48 ± 0.86 CFU/gut, respectively.Each donor-recipient pair was assayed for the ability to persist in the gut of flies for 1 h following inoculation with recipients. Donor and recipient strains were identified in the gut of s tested . The conectively . The rec1118 and CantonS flies  medium yielded no detectable transconjugants, indicating their absence in fly homogenates.In inoculations with the MM0002 donor and HS-4 recipient, no transconjugants were detected in either male or female W1118 or CantonS genetic background. Specifically, three male and female W1118 flies demonstrated transconjugant populations, of which one female fly demonstrated a population at a level higher than the theoretical LOQ. One male and six female CantonS flies demonstrated detectable transconjugants, with four female flies demonstrating transconjugant populations at levels higher than the theoretical LOQ.In inoculations with the MM0003 donor and HS-4 recipient, transconjugants were detected in a portion of both male and female flies of either the W1118 and CantonS flies were assayed for bacterial enumeration and conjugation efficiency following oral inoculation with donor-recipient pairs (S flies were colonized by recipient HS-4 at significantly lower (P < 0.05) concentrations than those of the recipient in female W1118 flies when inoculated with SP915 harboring either pKJK5-GM or pCVM29188_146 , no studies have yet used the laboratory research model Drosophila melanogaster as a host for gut-mediated HGT  pathway, or the Imd pathway, with the subsequent activation of both antimicrobial peptide secretion as well as the DUOX pathway  sequences, and replication origins  was used as a common donor for the plasmids pKJK5-GM, pCVM29188_146, and pC20-GM, referred to as MM0001, MM0002, and MM0003, respectively, throughout  plates supplemented with antibiotics selecting for chromosomal and plasmid resistance. Cultures were incubated overnight at 37°C prior to each assay, and purity was confirmed by the lack of resistance to alternate resistance markers.roughout . Briefly600) of ∼1.0, and 1 ml of the culture was centrifuged at 16,000 × g for 5 min and resuspended in 0.5 ml of LB broth. The donor and recipient strains were mixed at a 1:1 ratio in triplicate and incubated for 6 h at 25°C. Dilutions of the mating mixtures were plated onto MacConkey agar plates containing antibiotics selecting for the recipients and transconjugants, donors and transconjugants, or transconjugants only. The antibiotics used per conjugation are described in The ability of the donor strains to transfer AR plasmids was examined using a liquid culture mating technique as previously described . Initial600 of ∼0.8. Cultures were then centrifuged and resuspended in 200 μl resuspension buffer . Next, 400 μl of lysis buffer  was added, and cultures were mixed by inversion. Cultures were incubated at room temperature for 60 min. Following incubation, 600 μl of phenol-chloroform (1:1) was added, and cultures were mixed gently by inversion. Phases were then separated by centrifugation at 10,000 × g for 10 min at room temperature. The top aqueous solution was transferred to a new tube and used directly for 0.5% agarose gel electrophoresis as previously described  prepared according to the manufacturer’s instructions. Jazz mix medium consists of brown sugar, cornmeal, yeast, agar, benzoic acid, methylparaben, and propionic acid; however, specific concentrations of each component are not available. Prior to conjugation assays, flies were transferred to defined Bloomington Drosophila Stock Center (BDSC) cornmeal medium prepared without the addition of propionate  twice before suspension in 5% sucrose in sterile double-distilled water (ddH2O) to a final OD600 of approximately 100. For conjugation experiments, flies were fasted in clean empty fly tubes for a period of 4 h in groups of males or females (n = 12/group) 5 to 7 days after eclosion (ad libitum for 1 h. Recipient cell suspensions (100 μl) were likewise used to wet the cotton end of a new sterile swab and placed into a fresh BDSC fly medium tube. Flies were transferred to the recipient-inoculated fly tube and incubated for 1 h. Following incubation, flies were transferred to fresh BDSC fly medium tubes and incubated for one additional hour. All incubations occurred under standard Drosophila conditions as described above.Fresh cultures of donor and recipient strains  grown oveclosion . A 100-μ2 anesthetized, transferred individually into sterile microcentrifuge tubes, surface sterilized in 250 μl of 70% ethanol for 90 s, rinsed with sterile 1× PBS for 90 s, and then individually transferred to microcentrifuge tubes containing 100 μl of fresh sterile PBS. Bacterial populations of the gut were inferred from whole-fly bacterial enumeration following ethanol surface sterilization  were completed. To compare the mean ranks between groups, nonparametric Kruskal-Wallis analysis of variance (ANOVA) was used, and post hoc Dunn’s correction for multiple comparisons was applied. P values of ≤0.05 were considered significant.Statistical analysis was completed using the GraphPad Prism 6 software suite. Separate comparisons between sex  and genotype (W


Most studies either followed Joint National Committee 7 (JNC 7) or World Health Organization-International Society of Hypertension (WHO-ISH) guidelines to ascertain the prevalence of hypertension among Bangladeshi adults. The American College of Cardiology/American Heart Association (ACC/AHA) revised the definition of hypertension in 2017, which has significant public health importance. In Bangladesh, the new guideline has resulted changes in prevalence and risk factors for hypertension compared to the JNC7 guideline. This study used data from the most recent round (2017–2018) of the Bangladesh Demographic and Health Survey (BDHS). According to the 2017 ACC/AHA guideline, the participants were categorized as hypertensive if they had blood pressure (BP) ≥ 130/80 mmHg, but it was ≥ 140/90 mmHg in JNC 7 guideline. A total of 11,959 participants were involved in the analysis. The median (IQR) age of the respondents was 34.0 (18.0–95.0) years. The prevalence of hypertension was 24.0% according to the JNC 7 guideline, which was 50.5% according to the 2017 ACC/AHA guideline. Participants who were overweight and obese, aged, member of affluent households, Rangpur and Rajshahi division inhabitants had significantly higher odds of being hypertensive according to both guidelines. The new guideline suggests that half of the adult population in Bangladesh is hypertensive when measured according to the new guideline, urging the policymakers and public health practitioners to take immediate action to address the already established modifiable risk factors. What changes occurred in prevalence of and associated factors with hypertension among Bangladeshi adults (aged 18–95 years) due to 2017 ACC/AHA and JNC 7 guidelines?3. In 2016, around 17.9 million people died from CVDs, representing 31% of total global deaths, of which 9.4 million deaths were attributed to hypertension5. Worldwide, approximately 1.13 billion people have hypertension, and two-thirds of them live in low- and middle-income countries (LMICs), including Bangladesh4.Globally, cardiovascular diseases (CVDs) are considered the leading causes of deaths or disability-adjusted life years, where hypertension plays a pivotal role in CVDs7. Henceforth, hypertension remains the foremost disease burden among the major non-communicable diseases (NCDs) in Bangladesh, like other South Asian countries such as India, Nepal, Bhutan, and Sri Lanka8.Due to the recent epidemiologic and demographic transitions, Bangladesh has documented significant lifestyle and behavioral changes with an increased prevalence of hypertension. According to the 2011 Bangladesh Demographic and Health Survey (BDHS) and the 2010 Non-Communicable Disease Risk Factor Survey, the prevalence of hypertension among the adult population was 25.7% and 17.9%, respectively9. The 2017 American College of Cardiology/American Heart Association (ACC/AHA) Guideline for Prevention, Detection, Evaluation, and Management of High Blood Pressure in Adults reduced the BP threshold for hypertension. According to the 2017 ACC/AHA guideline, hypertensive individuals have a SBP of ≥ 130 mmHg and/or a DBP of ≥ 80 mmHg10. Therefore, previously considered prehypertensive participants are reclassified as hypertensive due to changes in the cut-off values13. For example, the prevalence of hypertension among the Nepalese adult population almost doubled following the 2017 ACC/AHA guidelines12.The blood pressure (BP) threshold to classify prehypertension and hypertension varies according to different guidelines. Previously, the Seventh Report of the Joint National Committee (JNC 7) on Prevention, Detection, Evaluation and Treatment of High Blood Pressure described hypertension as systolic blood pressure (SBP) of ≥ 140 mmHg and/or diastolic blood pressure (DBP) ≥ 90mmHg14. Another study found 45.1% and 26.8% increase in the prevalence of hypertension among adults (aged between 45 and 75) in the US and China15. Several studies recognized the importance of this revised classification for public health resource planning and prevention strategies16. Despite the increased burden of CVDs in developing countries, there remains inadequate information around the new guideline of hypertension19.Furthermore, Muntner et al. reported a 14.7% increase in hypertension among American adults over 19 years of age according to new guideline6. Islam et al. reported the findings from 1843 Bangladeshi adults aged over 18 years and found a similar increase (22.8%) in hypertension20. Besides, Kibria et al. concluded that the risk factors and their level vary between the JNC 7 and ACC/AHA guidelines11. Thus, changes in prevalence and risk factors have direct and indirect implications and hold significant merit in revising public health policies and plans to address the issue21. However, all these studies reported the findings from the 2011 BDHS survey data. A recent research article compared the prevalence and risk factors of hypertension between BDHS 2011 and BDHS 2017–2018 data using multiple logistic regression model22. The prevalence ratio (PR) is a suitable method for cross-sectional study when the prevalence of a disease is > 10%24. Hence, the statistical modeling in this study overestimated the estimated odds ratios for the risk factors. However, we applied appropriate statistical methods to estimate the PR, which is a uniqueness of this study.Previous studies in Bangladesh reported a significant increase in hypertension when the 2017 ACC/AHA guideline is being implemented to identify hypertensive individuals. Kibria et al. found that the prevalence of hypertension among Bangladeshi adults aged ≥ 35 years increased from 25.7 to 48.0%The present research was carried out to identify the change in the prevalence and associated factors of hypertension according to the ACC/AHA guidelines in comparison to that of the JNC 7 guideline using the most recent Bangladesh Demographic and Health Survey (BDHS) 2017–2018 data. This new dataset encompasses a comparatively larger measurement of BP with more participants than 2011 BDHS. Therefore, the findings using this latest dataset can broaden policy implications regarding hypertension management in Bangladesh.The study analyzed the most recent 2017–2018 BDHS dataset. The survey was carried out from October 2017 to March 2018 under the National Institute of Population Research and Training, Medical Education and Family Welfare Division, and Ministry of Health and Family Welfare. The survey’s principal objective was to assess the health indicators and provide an overview of population, maternal and child health, and the status of several NCDs such as hypertension and diabetes.25. A total of 12,152 unweighted sample was found in the original dataset, which increased to 12,975 after applying the weight to the dataset. However, we had to exclude 1016 cases due to missing values in some of the variables, making the total weighted number of observations 11,959 for our analysis.The sampling frame used for the 2017–2018 BDHS is the complete list of enumeration areas (EAs) covering the entire population of Bangladesh. The survey used a list of enumeration areas (EAs) provided by the Bangladesh Bureau of Statistics from the 2011 Population and Housing Census of the People’s Republic of Bangladesh. The survey’s primary sampling unit (PSU) is an EA covering on average 120 households in 2017–2018. The 2017–2018 BDHS was a multistage stratified cluster sample of households’ survey, carried out in two and three stages in rural and urban settings. In the first sampling stage, rural wards were selected, following PSUs, and then households were selected from PSUs. In urban areas, wards were selected through the PSUs technique, and one EA was selected from each PSU. Then, the households were chosen from the selected EAs sample. A detailed description of the survey design, methodologies, sample size, questionnaires, and findings is available in the final summary report of 2017–2018 BDHS. Anthropometry and BP were also systematically measured from the selected subsample of 2017–2018 BDHS® UA-767 Plus BP monitor at about ten minutes interval25. Then, the average of second and third measurements was used to report respondents’ final BP25. A person with a SBP of ≥ 140 mmHg and/or a DBP of ≥ 90 mmHg was considered hypertensive, as suggested by the JNC 726. While according to the ACC/AHA 2017 guideline, individuals with a SBP of ≥ 130 mmHg and/or a DBP of ≥ 80 mmHg or who were taking any prescribed antihypertensive drugs to control BP were categorized as hypertensive27. The category of prehypertension was transformed into elevated blood pressure in the 2017 ACC/AHA guideline27.The dependent variable for this study was hypertension. Trained health technicians measured BP three times using LIFE SOURCE31. The household factors included administrative divisions ; place of residence ; and wealth status , whereas the socioeconomic and individual factors included: age of the participants in years ; sex of the participants ; education level ; and occupational status . Behavioral characteristics included smoking habit  and body mass index (BMI) level. We have used global cut-off points for BMI classification: underweight (< 18.5 kg/m2), normal (18.5–25.0 kg/m2), overweight (25.1–29.9 kg/m2), and obese (≥ 30.0 kg/m2)32.The explanatory variables included in the study were selected based on previous literature reporting the risk of hypertension in LMICs setting® OnDemand for Academics for data analysis.Considering the complex survey of BDHS, we prepared the data using the survey weights before the analysis. Next, the normality assumption of continuous variables was investigated from their distribution, and it was reported in the paper with medians and interquartile ranges (IQRs). Then, we estimated the prevalence of hypertension and reported the differences between the two guidelines. We reported the prevalence of hypertension by background characteristics accounting for complex survey design/survey weight. It is notable to mention that survey weights only account for the sampling scheme. Therefore, we standardized the prevalence of hypertension for the same standard population to remove or minimize the impact of differences in participant’s age and sex distribution. On the next step, we fitted the selected explanatory variables in unadjusted log-binomial regression model. Thereafter, we executed a log-binomial regression model considering survey weights including the explanatory variables having p-values (< 0.05) in the unadjusted analysis to identify the factors associated with hypertension, describing results with prevalence ratios (PRs) and their 95% confidence intervals (CIs) and p-values. We used Statistical Package for Social Science (SPSS) version 26 and SAS25.The study used secondary data from the Demographic and Health Surveys (DHS) Program publicly available; therefore, we did not require any further ethical approval. The details of ethical procedures followed by the DHS Program can be found in the BDHS report2 (19.4–24.9). In addition, 25.4% had no education, and 73.4% were residing in rural areas.A total of 11,959 weighted participants were included in the analysis  prevalence of hypertension observed in Bangladesh was higher at the national level, while (20.9%) lower prevalence was documented in the previous recommendation of JNC 728. The plausible explanation could be biological and behavioural characteristics among the females might have increased over the period. This fact is supported by the previous evidence that females have a higher risk of obesity and diabetes compared with men37. This needs females to require more awareness and public health information to control hypertension and minimize adverse complications28. Our study findings reported that people with higher socioeconomic status had higher odds of having hypertension. The higher wealth status participants can generally purchase more consumable resources with a large amount of calorie intake, making them overweight or obese, putting them at a greater risk of being hypertensive than those lower wealth status39. This suggests the need for prevention and control program for hypertension in urban areas of Bangladesh.Interestingly, our findings reported a similar prevalence of hypertension among male and female participants. If we compare the finding based on the previous study, the prevalence rate increased among female participants more than male counterparts41. The possible reason could be prevailing unhealthy lifestyle factors such as less physical activity, consumption of unhealthy diets among the urban populations might have contributed to the disease burden44. However, this finding warrants further detailed investigation of causes for the increased prevalence or odds of hypertension in several Bangladesh divisions28. This finding suggests the need to understand the social inequalities among the rural and urban community, which may have played a role in such variation. Understanding the inequalities mentioned earlier may help design the comprehensive hypertension prevention and control program for Bangladesh peoples.The prevalence of hypertension was higher among those living in urban areas, which is in line with previous studies where urban people were reported more hypertensive28. Since most urban participants are educated, and these had a higher prevalence of hypertension. Thus our study recommends that educated individuals in urban areas need to receive more public health awareness information to control raised blood pressure levels28.There is another explanation that would help understand why the prevalence of hypertension is high in urban areas. The study found that higher educated and higher wealth status of people are likely lives in urban areas, resulting from having a sedentary lifestyle such as low physical activity. A lack of open spaces for playing games or physical activity might result in the high-risk prevalence of hypertension47. Notably, in the current study, administrative divisions were also found significantly associated with hypertension in line with suggest 2017 ACC/AHA guidelines. People from Rangpur and Rajshahi division found higher odds in the two guidelines. Much is unknown why the people from these two divisions owned higher risk of being hypertensive; however, the reason may be because of socioeconomic inequalities such as limited resources, income inequality, low level of education and social safety net programs, poor connectivity with the urban centres, insufficiency or absence of public infrastructure49.This study identified the potential risk factors of hypertension using both JNC 7 and 2017 ACC/AHA guideline alongside to the estimation of the prevalence. People of older age 25 to more, overweight, and obese had relatively higher odds, which is in line with previous studies elsewhere12. Our study findings are significant because it shows that above fifty per cent of adults with hypertension or elevated blood pressure according to the new 2017 ACC/AHA classification require active lifestyles and healthy dietary habits. Public health programs should adequately address this emerging problem; in Bangladesh, emphasis should be paid to prevention and self-management of a condition not only for those with hypertension but also for all adults12. Therefore, it is essential to estimate the prevalence based on both thresholds to control this hypertension burden, which might exacerbate cardiovascular disease. These findings might help future researchers and appropriate authority design any programs and policies regarding control and prevent hypertension burden and overcome this massive public health challenge.The new 2017 ACC/AHA guideline recommends treating stage 1 hypertension with changing lifestyle measures and taking antihypertensive medicines to prevent future cardiovascular disease risksThe strengths and weaknesses of this study are accredited. The strength that lies in this study is the generalizability of the findings for Bangladesh since this survey covered nationally representative data covering all divisions. Along with appropriate statistical methods to estimate the weighted prevalence of hypertension from the sample.12. This survey also used an automated device, though both guidelines recommend recording blood pressure with a sphygmomanometer51.The limitations of the study are appropriately acknowledged. Due to a cross-sectional setup, no causality cannot be established, and the individuals’ blood pressure was measured three times in a single day. However, both guidelines recommend longitudinal measurement of blood pressure levels to diagnose hypertensionThe present study highlighted that the prevalence of hypertension was almost doubled according to the 2017 ACC/AHA guideline compared to the JNC 7 guideline. The policymakers and public health practitioners should consider the new guideline and make new strategies to increase awareness among the adult population in Bangladesh. The study finding also points towards addressing the already established modifiable risk factors of hypertension such as overweight/obesity, high-income status, which are also identified as the risk factors according to both guidelines.Supplementary Table 1.


Oryza sativa L.) cultivar Liaojing 371 was used. Compared with the plants in CK, those in the FM treatment showed greater average root diameter, root volume and root dry weight. The SF treatment increased the single stem flexural strength, increased the contents of silicon, lignin, and cellulose in the rice plant stalk, and reduced the lodging index, thereby increasing lodging resistance. The SF treatment resulted in increased leaf chlorophyll content at late growth stage and a higher net photosynthetic rate, which increased plant dry matter accumulation. In the FM + SF treatment, plant growth was enhanced during the whole growth period, which resulted in an increased number of effective panicles and an increased grain yield. The results show that the combination of FM and SF synergistically improves rice lodging resistance and grain yield. This low-cost, high-efficiency system is of great significance for improving the stability and lodging resistance of rice plants, thereby increasing yields.The use of bast fiber film can improve rice seedling quality, and nano-silicon fertilizer can increase rice yields. This study aimed to compare the effects of using bast fiber film, nano-silicon fertilizer, and both treatments on rice yield and lodging resistance. A 2-year field experiment was conducted in 2017 and 2018, in Liaoning, China. The experiment comprised a control , and three treatments: seedlings cultivated with bast film (FM), single nano-silicon fertilization (SF), and bast fiber film seedlings + nano-silicon fertilization (FM + SF). The japonica rice ( Bast fiber film has been widely used in agriculture in China since 2015 because it is inexpensive, improves seedling quality and uniformity, reduces seedling losses from seedling plates, and ensures better mechanical transplanting3. Compared with ordinary plastic film, bast fiber film provides a more constant temperature environment for rice seedlings5. It is also breathable and hygroscopic. Mulching with bast fiber film on the seedling tray increases the oxygen supply to the soil so that oxygen consumed by soil microbial activity is readily compensated6. Using bast fiber film to raise rice seedlings can promote root growth and enhance seedling vitality, and the stronger root systems improve seedling quality. After transplantation into the field, the rice seedlings cultivated with bast fiber film rapidly accumulate photosynthetic pigments and produce tillers early, which increases the number of effective panicles8. In addition, the bast fiber film is degraded into organic matter, which has a soil-fertilizing effect and thus further stimulates the growth and development of transplanted rice9. There is a close and direct relationship between the rice root system and the photosynthetic characteristics of aboveground leaves10. In rice plants, rapid pigment accumulation and root growth and development facilitate photosynthesis. This leads to the accumulation of photosynthates in the aboveground parts of rice plants at an early stage of growth, so that more resources are available for grain filling.Bast fiber film is an eco-friendly and biodegradable seedling raising cloth that is made from bast fibers and small amounts of eco-friendly biodegradable pulp10. Generally, the SiO2 content in stems and leaves reaches 10%–20% on a dry weight basis, so rice is a representative siliceous plants11. Silicon plays an important role in rice growth, and large amounts of this mineral are required to maintain normal growth and development12. The application of silicon fertilizer can increase chlorophyll content and delay leaf senescence in the late growth stage of rice, thus increasing photosynthesis in the functional leaves13. This increases the ability of rice plants to assimilate photosynthetic products during the vegetative growth stage, which improves grain filling and grain quality, and increases resistance and yield16. Silicon fertilizers can also increase the cellulose content in rice plants during the whole growth period, which in turn hardens the stems and enhances resistance to the lodging and certain diseases. Nano-silica is a novel type of nano-scaled silicon fertilizer made from particles of sparingly soluble materials by nanotechnology methods17. Small particle-sized nano-fertilizers are more likely to pass through biological barriers, such as plant cell walls, to fully exert their effects18. To date, few studies have evaluated the effects of nano-silicon fertilizers on rice plants.As a typical silicon-accumulating crop, rice plants contain large amounts of silicon. The total silicon content in rice plants is greater than the sum of the contents of nitrogen, phosphorus, and potassium5. However, it also results in increased plant height, which increases the risk of lodging19. At present, little is known about the effect of bast fiber film on rice plants at the later stages of growth and development. The middle to late stages of grain filling are crucially important for rice yield, and the photosynthetic characteristics in the critical growth period of rice and lodging at later stages determine the magnitude of yield23. Nano-silicon fertilizers can delay senescence and enhance the resistance of rice plants24. Therefore, we speculated that the combination of the blast fiber film and nano-silicon fertilizer might synergistically affect both rice lodging resistance and yield. If so, then this would be a low-cost, simple, and effective way to improve rice production.Previous studies on the use of bast fiber film to raise rice seedlings have reported its beneficial effects on the growth and development of rice at the seedling stage and after transplantation−1; total nitrogen content, 1.34 g/kg−1; available nitrogen content, 85.68 mg/kg−1; available phosphorus content, 12.74 mg/kg−1; available potassium content, 90.99 mg/kg−1; and available silicon content, 176.64 mg/kg−1.A field experiment was conducted in 2017 and 2018 at the Rice Research Institute in Liaoning Province, Shenyang, Liaoning, China . This area is in the North Temperate Zone and has a semi-humid continental climate, with an average annual temperature of 8 °C, an average annual precipitation of 660 mm, and an average annual frost-free period of 170 days. The basic physical and chemical properties of the paddy soil (0–20 cm) were as follows: pH 6.87; organic matter content, 25.13 g/kgjaponica rice variety Liaojing 371 was used. The bast fiber film used in this study was developed by the Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, and the fertilizer was ‘Silicon Boyuan’ micro-nano-silicon fertilizer. The experiment comprised a control  and three treatments: seedlings raised with bast fiber film (FM), a single nano-Silicon fertilization treatment (SF) and bast fiber film + nano-silicon fertilization (FM + SF). Seeds were sown uniformly with an automatic rice planter at 100 g per tray (60 cm × 30 cm) in soil. Ten seedling trays were established for each treatment. The trays were randomly placed in a greenhouse and managed in the same way as in a conventional dry nursery. Seven days before transplanting, an aqueous solution of micro-nano-silicon fertilizer  was evenly applied to seedlings in the SF and FM + SF treatments. The seedlings were transplanted at the 3.5-leaf age at a row spacing of 30 cm × 16.7 cm. The planting area of each plot was 100 m2. Each treatment had three replicates and the experiment had a randomized block design. Seeds were sown on April 25th, seedlings were transplanted on May 25th, and the mature plants were harvested on October 1st.The conventional Samples were taken from the tray one day before transplanting. Ten representative rice seedlings in each plot were sampled to measure root length and root volume using a root scanner and supporting analytical software . At maturity, The dry weight of aboveground parts was determined and five representative plants were selected for analysis(The average number of tillers of ten consecutive rice plants is used as a representative standard). The plants were divided into four parts: ear, stem, leaf, and sheath. The parts were dried separately in an oven at 105 °C for 10 min, and then at 80 °C to constant weight.The upper third of the rice flag leaves was used to measure the SPAD value and photosynthetic characteristics on the 15th day after full heading. The SPAD value and photosynthetic characteristics were measured with a SPAD-502 chlorophyll meter  and an LI-6400 portable photosynthesis system , respectively. The photosynthetic characteristics were net photosynthetic rate (A), stomatal conductance (Gs), intercellular carbon dioxide concentration (Ci), and transpiration rate (E). These measurements were conducted between 9 and 11 am on a sunny day.25. The stems were collected from mature plants. The silicon concentration in the plant stem was determined by the colorimetric molybdenum blue method26, and cellulose and lignin contents were determined using Van Soest’s washed fiber analysis method27.On the tenth days before maturity, rice plants from five hills per plot were selected, five representative tillers with consistent growth were chosen using the secondary sampling method, and the average value was determined. The plant height, center of gravity height, basal internode (including leaf sheath) length, stem diameter, and dry matter weight were recorded. The internode breaking resistance was measured with a stem strength tester 2 per treatment. After drying and threshing, the rice grains were weighed to calculate grain yield. The number of effective panicles in 20 consecutively growing rice plants in each treatment was counted. Use the average number of tillers as the standard for selecting rice to determine the yield components. The number of grains per panicle, seed setting rate, 1000-grain weight and the grain yield were recorded.At harvest, the rice yield was measured for an area of 10 mData were processed and analyzed using GraphPad Prism 8 and SPSS 18 statistical software. Significant differences among treatments were detected using two-way ANOVA.Oryza sativa L (either cultivated or wild), including the collection of plant material, comply with relevant institutional, national, and international guidelines and legislation.My experimental research and field studies on Both the FM and SF treatments had significant positive effects on the number of panicles Table . The numThe root projected area, root surface area, and root volume differed significantly among treatments Table . The 2-yThe A and Gs values were 19.1% and 16.9% higher, respectively, in the FM + SF treatment than in CK. There were no significant differences in Ci and E among treatments. The SPAD values were significantly higher in FM + SF and SF treatments than in CK. The A, Gs, E, and SPAD values of plants in the various treatments are shown in Table As shown in Table As shown in Table As shown in Table −1 and 6.74 mg.g−1, respectively, 38% and 34% higher than that in CK (Fig. The application of nano-silicon fertilizer significantly increased the silicon content in rice stems. The stem silicon content in the FM + SF and SF treatments was 6.94 mg.g CK Fig. .Figure 2The cellulose and lignin contents in stems were also significantly higher in the SF and FM + SF treatments than in CK Fig. . The cel28. The use of bast fiber film benefits the root growth of rice seedlings. It can promote root primordia differentiation, which increases the numbers of adventitious roots and denser and better-quality root networks29. Mulching with bast fiber film can also increase rice seedling root vitality and the number of white roots, which contributes to whole plant growth. This leads to improved seedling quality and faster pigment accumulation after transplanting30. Higher dry matter accumulation in above-ground parts of seedlings can induce earlier tillering after transplanting, which resulted in an increased number of effective tillers and higher yields33. The results of this study show that the use of bast fiber film can promote the growth and development of rice seedling roots. In this study, seedlings cultivated with bast fiber film showed increased dry weight, mean diameter, volume, and surface area of roots, increased stem base diameter and aboveground growth, and improved plant quality. These attributes resulted in rapid greening after transplanting and better growth and development of photosynthetic organs. These effects resulted in rapid dry matter accumulation, which provided the basis for higher and more stable rice yields.Because bast fiber film is a breathable mulch, oxygen consumed at the bottom of rice seedling discs is readily replaced, similar to the case in aerobic cultivation34. In this study, the use of bast fiber film to cultivate seedlings and the application of nano-silicon fertilizer at the seedling stage increased the chlorophyll content of rice leaves and delayed senescence, thereby enhancing photosynthesis in functional leaves. This improved the capacity of rice plants to assimilate photosynthetic products during the growth stage. Our results also show that the use of bast fiber film to cultivate seedlings resulted in increased dry matter accumulation in leaves and stems, thereby increasing biological and grain yields. Using bast fiber film to cultivate seedlings or applying nano-silicon fertilizer at the seedling stage slightly increased yield compared with that of the control, but the combination of these two treatments increased the effective panicle number per unit area, which significantly increases the yield.The SPAD value is an indirect index of leaf chlorophyll contentIn the rice plants in the combined bast fiber film and nano-silicon fertilizer treatment, the flag leaf chlorophyll content at the late growth stage was 4.8% higher, the net photosynthetic rate was 10.9% higher, and the dry weight at the mature stage was 9.5% higher than their respective values in the control. Our results show that the use of bast fiber film can promote seedling regeneration after transplanting and that the application of silicon fertilizer can delay senescence, enhance the photosynthetic capacity, and increase dry matter accumulation, thereby boosting yields.35 reported that a high lignin level in the cell wall improves lodging resistance, and suggested target genes for the genetic modification of lignin content to breed rice lines with high lodging resistance. Our results show that the application of nano-silicon fertilizer can significantly increase the contents of silicon, lignin, and cellulose in stalks, shorten internode length, and reduce plant height. All these changes increase lodging resistance. In addition, the application of nano-silicon fertilizer can significantly increase the dry weight per unit length, which lowers the lodging index and increases lodging resistance.Liu et al.Using bast fiber film to cultivate rice seedlings can promote growth at the cost of increasing the risk of lodging. Applying nano-silicon fertilizer can delay senescence and improve the lodging resistance of rice plants. Our results show that the combination of bast fiber film to cultivate rice seedlings and nano-silicon fertilizer synergistically affects rice yield, lodging resistance, root characteristics, and leaf photosynthetic traits. Thus, this is a low-cost and simple method to improve rice yields effectively and efficiently.


Background: The results of previous studies are heterogeneous about the effect of body fatness on risk of gastric cancer (GC). Herein we investigated the effect of changes of BMI and body shape on risk of GC.Methods: A population-based case-control study enrolled 1989 controls and 937 GC cases. Logistic regression models were used to calculate odd ratios (ORs) and 95% confidence intervals (CIs) for BMI and body shape in association with GC risk, according to anatomical subsite, Laurén's classification, sex and Helicobacter pylori (Hp) infection.Results: Subjects with higher BMI or body shape 10 years before interview had a lower risk of GC regardless of anatomical subsite, Laurén's classification, and sex . But the relative risk patterns were different by Hp status. When checking the effect of changes of body fatness, in Hp+ stratum, the ORs (95% CI) were 0.40 (0.17-0.93) for subjects who were underweight at age 20 but had increased BMI afterwards, and 0.48 (0.32-0.73) for those of body shape 1/2 at age 20 but increased body shape subsequently, compared to subjects with stable BMI or body shape. When subjects had a normal BMI or 3/4 body shape at age 20, weight loss nearly doubled the risk of GC, and weight gain would decrease the risk.Conclusion: The association between body fatness and GC risk might differ by time point of measurement and Hp-infection status. Further, the influence of changes of body fatness might be different by baseline body fatness and Hp-infection status. Helicobacter pylori (Hp) infection Gastric cancer (GC) is one of the most dreadful malignancy around the world, with over 1,000,000 new cases and an estimated 783,000 deaths reported in 2018 et al found that adolescent obesity was associated with an increased risk of subsequent TGC in both men and women et al pointed out that higher body mass index (BMI) was associated with a decreased risk of TGC, particularly in men and older persons GC can be subdivided by anatomic subsite into esophagogastric-junction cancer (EGJC) and true gastric cancer  BMI is one of the most used measures to assess body fatness, and it has high specificity but low sensitivity to identify adiposity Taixing of Jiangsu Province, situating in Eastern China, is a high incidence area of GC The detailed design of this population-based case-control study could be found in our previous publications Eligible participants were limited to those between ages of 40 and 85 years, and living in Taixing for at least 5 years. GC cases were recruited if the patients were suspected as GC cases during gastroscopy examination in four largest local hospitals , where more than 90% of the patients in Taixing were diagnosed. And we established a rapid case recruitment system that directly notified our study staff to interview the suspected GC cases. To find all potential GC cases, a cross-matching with the local Cancer Registry's list of cases was made each year to identify any missing cases. Finally, all cases were examined by a pathologist and excluded if not confirmed. The detailed process is shown in Figure Controls were from the local Population Registry System, which covers the whole population of Taixing. They were randomly selected every 12 months and a frequency matching method for control selection was used to increase statistical power. We recruited the controls at the same time for all upper gastrointestinal cancer cases because of the approximately identical distribution of age and sex. All upper gastrointestinal cancer cases were stratified by sex and 5-years age groups, and corresponding controls were selected by an appropriate ratio for each stratum, considering the response rate among controls. In this way, 3501 controls were randomly selected, of whom 643 were excluded because of the death before being contacted, outmigration or inability to be contacted. Finally, 2011 controls participated in this study (response rate: 70.4%), of whom 19 controls were out of the age range (40-85 years) and 1992 controls were included in the final dataset.2; normal, ≥ 18.5 to < 24 kg/m2; overweight, ≥ 24 to < 28 kg/m2; obesity, ≥ 28 kg/m2. Body shape was chosen from the revised image of somatotypes . In this study, Hp infection was defined as Yes if the Ure-B antibody was seropositive .All participants were interviewed by trained interviewers face to face, using an electronic questionnaire. Cases were under investigation before they knew their diagnoses of cancer. Data on demographic characteristics and other study variables were obtained, including the body weight, height and body shape at age 20 and 10 years before interview. BMI was calculated as the body weight in kilograms divided by the square of the height in meters and divided into four categories: underweight, < 18.5 kg/matotypes  25, fromAnatomically, GC cases were divided into EGJC and TGC based on the illustration of tumor sites in their gastroscopy reports. Meanwhile, GC cases were also classified as intestinal type and diffuse type, according to Laurén's classification ORs) with 95% confidence intervals (CIs) for BMI and body shape in association with risk of GC, according to anatomical classification, Laurén's classification, sex or Hp infection, respectively. All relative risk estimates were adjusted for the following potential confounding factors: age (continuous), sex , education , marital status , occupation , Hp infection (yes or no), sum of missing and filled teeth , daily frequency of brushing teeth (< 2 or ≥ 2), tea drinking (yes or no), smoking , alcohol drinking , job intensity , family wealth score (quintiles) and family history of GC among first-degree relatives (yes or no). Likelihood ratio tests were used to test the effect modification by family wealth, smoking, alcohol drinking and job intensity. Then, sensitivity analyses were undertaken by excluding cases only from the local Cancer Registry and by excluding subjects older than 75. All analyses were performed using Stata software (version 14.0). All tests were two-sided and P-values less than 0.05 were considered significant.Distribution of demographic characteristics and other study variables was presented as mean and standard deviation for continuous variables, and frequency and percentage for categorical variables. Unconditional logistic regression models were used to calculate odds ratios . This pattern also applied to body shape 10 years before interview, and a more obvious relative risk gradient was observed. Analysis by anatomic subsite showed similar results (The associations between BMI and body shape in different time points (age 20 and 10 years before interview) and risks of GC are shown in Table  results , and so  results  and sex  results .Hp infection. In the Hp+ stratum, compared with those with normal BMI 10 years before interview, underweight subjects had an increased risk of GC, while overweight or obese subjects had a lower risk of GC (P for trend < 0.001). The relative risk pattern was similar for body shape 10 years before interview . No significant associations were found between BMI or body shape at age 20 and the risk of GC. However, in the Hp- stratum, relative risk patterns were different. Although there was no clear trend of ORs in association with BMI, higher body shape at age 20 was associated with an increased risk of GC (P for trend = 0.018). Compared with those with body shape 3 10 years before interview, ORs was 2.62  for body shape 1, and 2.12  for body shape 5.Table Hp infection  compared with those keeping underweight all the time. Compared with those who had a normal BMI both at age 20 and 10 years before interview, those with decreased BMI 10 years before interview had an almost doubled risk of GC , while those with increased BMI had a 41% decreased risk of GC . For change in body shape, the relative risk patterns were almost similar. Compared with those with stable body shape 1/2, those with body shape 1/2 at age 20 but higher body shape 10 years before interview had a more than 50% decreased risk of GC . And compared with those with stable body shape 3/4, those with lower body shape later had an almost doubled risk of GC . In the Hp- stratum, the direction of most point estimates was consistent, although almost all results were not statistically significant, except that those with lower body shape later had an increased risk of GC  compared with those with stable body shape 3/4 from age 20.We further explored the effect of change in BMI and body shape stratified by on Table . In the p-value of interaction >0.05). Finally, sensitivity analysis was respectively performed by excluding 60 male cases and 21 female cases collected only from the local Cancer Registry, and by excluding subjects older than 75. The results did not change materially (data not shown).Besides, analyses of possible effect modification by family wealth, smoking, alcohol drinking, and job intensity were performed. The results were consistent and likelihood ratio tests did not detect any statistically significant interaction  2) had a higher risk of overall GC et al found a higher risk of EGJC with increasing BMI at age 20 et al found the same trends for TGC It remained controversial for the association between body fatness and risk of GC, when GC was separated into EGJC and TGC by anatomical subsite Meanwhile, we explored two other subtypes, intestinal type and diffuse type, according to Laurén's classification Hp was classified as a Group I carcinogen for GC by the International Agency for Research on Cancer (IARC) Hp infection. The protective effect of higher BMI 10 years before interview was just found in the Hp+ stratum. A possible explanation was that weight loss could be caused by Hp infection. Lender et al found a negative association between Hp infection and BMI level Hp infection could cause a series of gastric diseases, such as chronic gastritis Hp, subjects might suffer from chronic gastritis and other gastric diseases for a long time, even more than 10 years, which could cause their weight loss by affecting food intake or digestion.Hp infection, those underweight or with normal-BMI (BMI < 24 kg/m2) at age 20 had a higher risk of GC in association with a decreased BMI 10 years before interview, and a lower risk with gain in BMI. There were few studies considering the effects of changes in body fatness et al calculated as BMI at baseline minus BMI at age 20 years and found that subjects with a BMI gain of >8.0 kg/m2 had a 2.07 times higher risk  of EGJC compared to subjects with 0-3.9 kg/m2 change in BMI et al only found modestly association of weight change with EGJC In our study, among subjects with et al to depict their body shape Hp- stratum. It was found that higher body shape at age 20 was positively associated with risk of GC, which was consistent with the findings of Merry et al and Wu et alThere were other commonly used measurements of body fatness besides BMI, such as abdominal diameter Discrepancies between our results with those of previous studies might be due to different study populations. Several studies in Asian countries showed a borderline significant or no associations between BMI and risk of GC The biological mechanism is unclear for the association between body fatness and risk of GC. It has been realized that adipose tissue is not only a reservoir for energy storage, but also an essential metabolic and endocrine organ Besides the use of both BMI and body shape to assess body fatness, there are some other strengths in our study. First, we explored the associations for different classifications of GC. Second, our trained staff interviewed cases before they knew their cancer diagnoses, in order to control recall bias. Furthermore, all cases were carefully verified by a pathologist and a high response rate for both cases and controls was also the strength.Our study also has several limitations. We did not gather information of BMI and body shape at more time points, so that the current results only indicate the association between a relative long-term change and risk of GC. Furthermore, recall bias cannot be absolutely eliminated. But we made efforts to control it in aforementioned ways and found no material change when conducting a sensitivity analysis by excluding cases only from the local Cancer Registry.Hp infection status. Further studies are needed to clarify whether the observed effect of body fatness on GC risk is due to the development of precancerous lesions, or on the pathway between Hp infection to cancer development, or other to-be-proven mechanisms.In summary, body fatness is associated with GC risk, especially in the time point closer to cancer development. Changes of body fatness also have differential effects on GC risk by baseline body fatness and Supplementary figures and tables.Click here for additional data file.


After seven days of exposure, lung function testing was performed, and lungs were taken for stereological analysis. In addition, the ratio between pulmonary arterial acceleration and ejection time (PAAT/PAET) was measured. Inspiratory capacity and static compliance were reduced whereas tissue elastance and resistance were increased in hyperoxic animals compared with normoxic controls. Hyperoxic animals showed signs of pulmonary hypertension indicated by the decreased PAAT/PAET ratio. In hyperoxic animals, the number of alveoli and the alveolar surface area were reduced by one-third or by approximately 50% of control values, respectively. However, neither the mean linear intercept length nor the mean alveolar volume was significantly different between both groups. Hyperoxic pups had thickened alveolar septa and intra-alveolar accumulation of edema fluid and inflammatory cells. Nonparenchymal blood vessels had thickened walls, enlarged perivascular space, and smaller lumen in hyperoxic rabbits in comparison with normoxic ones. In conclusion, the findings are in line with the pathological features of human BPD. The stereological data may serve as a reference to compare this model with BPD models in other species or future therapeutic interventions.Bronchopulmonary dysplasia (BPD) is a complex condition frequently occurring in preterm newborns, and different animal models are currently used to mimic the pathophysiology of BPD. The comparability of animal models depends on the availability of quantitative data obtained by minimally biased methods. Therefore, the aim of this study was to provide the first design-based stereological analysis of the lungs in the hyperoxia-based model of BPD in the preterm rabbit. Rabbit pups were obtained on gestation day 28 (three days before term) by cesarean section and exposed to normoxic (21% O Bronchopulmonary dysplasia (BPD) is a chronic human lung disease associated with preterm birth. In the early days, BPD was described to be the result of a combination of respiratory distress syndrome and the effects of therapeutic interventions, including ventilation and oxygen supply . With thth to 30th gestational week in humans [For these purposes, preclinical experiments addressing the pathophysiology of arrested lung development and the effects of new therapeutic interventions are essential , 10. Curn humans , 13. Hypn humans . Howevern humans . In contn humans . These mn humans . As a brn humans , 18.Morphometric methods are extremely valuable to characterize the lungs of different animal models in quantitative terms , 20. At th day of gestation , the does were sedated with intramuscular (i.m.) medetomidine 0.25 mg/kg . Ten minutes later, they received i.m. 20 mg/kg of ketamine  and 5 mg/kg of xylazine . Subsequently, the does were euthanized with an overdose of pentothal sodium . After death occurred, the abdomen was immediately opened, and the uterus exposed to extract all pups through hysterectomy. At delivery, pups were dried, stimulated, and divided into two groups similarly as described by Salaets et al. [Time-mated New Zealand white rabbits were obtained from Charles River Laboratories (France). All experimental procedures were approved by the local animal ethics committee and complied with the standard European regulations on animal research (no. 744/2017). For cesarean section at the 28s et al. : the norAfter 7 days of exposure with normoxic or hyperoxic air, the pups were anesthetized with isoflurane (2.5% in pure oxygen) and were placed in the supine position on the Vevo Imaging station . Two-dimensional images of the pulmonary artery were obtained from a parasternal short axis view at the level of the aortic valve using the MX-550D transducer . Measurements were performed offline (Vevo LAB software package V3.0). The following variables were measured: pulmonary artery acceleration time (PAT), ejection time (PET), and PAAT/PAET ratio. An average of three cardiac cycles was used for the analyses.At the end of the experimental period, pups were anesthetized with ketamine (35 mg/kg) and xylazine (6 mg/kg) i.p., and lung function testing was performed using a forced oscillation technique with the flexiVent system  . The ana2O with a fixative containing 1.5% glutaraldehyde and 1.5% paraformaldehyde in 0.15 M HEPES buffer. Our inflation fixation protocol was tested and optimized in advance for inflation fixation in preterm rabbits. After at least 24 h submersion in fixative, lungs were further processed for stereological analysis. Prior to embedding in Technovit resin, lung volume measurements were assessed by volume displacement (based on Archimedes' principle), and lung tissue was sampled by applying systematic uniform random sampling [μm thick sections and stained with toluidine blue or into two sequential sections with a distance of 4.5 μm thickness and stained with orcein for disector analysis and alveolar counting [Lungs of preterm born rabbits were fixed by instillation at a pressure of 25 cmHsampling . This wasampling , 30. Rescounting . HistoloStereological analysis was done with the newCAST™ software  on digitalized slides. For the estimation of parenchymal, nonparenchymal, and ductal airspace volumes, automated systematic uniform random subsampling (SURS) was performed at a 5x objective lens magnification and with a sampling fraction of 5-7% to collect the images for further analysis. A point grid consisting of 36 points with a 4-fold subsampling fraction was superimposed over the collected images and used to estimate the respective volume densities as described previously in detail . Respectμm was superimposed over the images, and points and intersections were counted. Respective volume and surface densities were estimated as described previously [For the estimation of alveolar and ductal airspace, septal volume, septal thickness, and septal surface area, an automated SURS was performed at a 20x objective lens magnification and with a sampling fraction of 0.5-1%. A test line grid with 12 lines and a length per point of 9.37 eviously , and totμm2 was superimposed over the disector image pairs, and the number of alveoli was estimated as described previously [For the estimation of the alveolar number, the consecutive tissue sections were aligned in the Visiopharm software. An automated SURS was performed at a 20x objective lens magnification with a sampling fraction of 0.9-1.8%, resulting in approximately 100 to 140 image pairs per lung. A counting frame with an area of 35,570.40 eviously , 32. Meaeviously .μm was used to calculate the vascular volume and surface area in vessels with a diameter of more than 25 μm. A point grid with 16 × 16 points was used to estimate the mean volume and thickness of the perivascular area and vascular wall in vessels with diameter of more than 25 μm. Calculations were done as described previously [The vascular volume, vascular wall thickness, and perivascular area were estimated using a sampling fraction of 4-9% with approximately 170-200 images at a 10x objective lens magnification. A length grid with 12 × 9 lines and a length per point of 18.74 eviously .t-test using the SigmaPlot® software (SYSTAT® Software Inc.). If normality testing failed, the Mann–Whitney rank sum test was used. Results were considered significant if p ≤ 0.05.A number of 9-10 rabbits per experimental group were included in the study. Lungs that did not properly inflate during organ harvesting due to technical issues were excluded from morphometric analysis, resulting in stereological analysis of 8 lungs per experimental group. Statistical comparison between the two groups was done with Student's The pups were weighed, and pulmonary artery microultrasound Doppler analysis and lung function measurements were performed prior to sacrifice. The average body weight did not differ between the two experimental groups after 7 days of exposure to hyperoxic or normoxic air and was 50 g (±4.9 g) for the normoxia-exposed pups and 49 g (±9 g) for the hyperoxia-exposed pups. However, significant differences were found in inspiratory capacity (IC), static compliance (Cst), tissue resistance (G), and tissue elastance (H) between the two experimental groups Figures . While IThe lung volume, assessed with volume displacement after instillation fixation, was also significantly reduced with exposure to hyperoxia  — similaμm  and septal edema and/or fibroproliferation . In addition, the luminal volume of nonparenchymal blood vessels was reduced whereas the volume of their walls and perivascular connective tissue space was enlarged. Functionally, the lungs showed significant signs of parenchymal damage (increase in tissue resistance and elastance) in the absence of strong effects on larger airways (similar airway resistance). However, signs of pulmonary hypertension were present (decreased PAAT/PAET ratio).Design-based stereology is the current gold standard of lung morphometry  because Most of the data reported in the current study are in accordance with previous morphometric studies in this animal model , 26 whic2 and 130-140 m2 depending on whether the estimation based on light microscopy/microcomputed X-ray tomography [Alveolar surface area is an important parameter to evaluate the functionally relevant diffusion area for gas exchange . Its estmography  or electmography . Tissue mography , 43.2 resulted in a significant decrease in alveolar number whereas both total alveolar surface area and mean linear intercept length were not significantly different from those of the control group [The most straightforward analysis of alveolarization is to estimate the total alveolar number. While originally based on geometric assumptions , an effiol group . In a syol group  providedIn the present study, the alveolar number was reduced to 68% of control lungs with no changes in Lm or mean alveolar volume, which corresponds well with the data for P4 to P7 hyperoxia exposure in the mouse study by Nardiello et al. . In contμm in diameter were grouped into a single compartment because previous studies using 3D reconstructions of the vasculature have shown that it may be impossible to distinguish morphologically between small arteries and veins in single 2D sections [Besides the parenchymal changes, the present study also provides evidence for an involvement of larger sized arteries and veins in the hyperoxia model of BPD. Arteries and veins larger than 25 sections , 53. Nevsections .The present study provides robust stereological reference data for the preterm rabbit hyperoxia model of BPD. The data confirm and extend previous morphometric studies in this animal model and fit well with the pathological features of human BPD. In contrast to their normoxic counterparts, the lungs of preterm rabbits exposed to hyperoxia were characterized by reduced alveolarization, septal edema and/or fibroproliferation, thickened vessel walls, functional impairment of the parenchyma, and pulmonary hypertension. Together, these findings may serve as a reference to compare this model with BPD models in other species or future therapeutic interventions.


How do birds navigate their way? It is one of the interesting question about homing pigeons, however the genetic of navigation has reminded as a puzzle. Optic lobe, olfactory bulb, hippocampus and cere were collected for RNA sampling. The generated RNA-seq represent RNA resequencing data for racing homer (homing) pigeon and other rock pigeon breeds. The obtained data set can provide new insight about hippocampus role and GSR contribution to pigeon magnetoreception.To investigate the navigation ability of rock pigeon breeds, 60 whole transcriptome sequence data sets related to homing pigeon, Shiraz tumblers, feral pigeons and Persian high flyers were obtained. RNA extraction was performed from three brain regions  and cere. Paired-end 150 bp short reads (Library size 350 bp) were sequenced by Illumina Hiseq 2000. In this way, about 342.1 Gbp and 130.3 Gb data were provided. The whole transcriptome data sets have been deposited at the NCBI SRA database (PRJNA532674). The submitted data set may play critical role to describe the mechanism of navigation ability of rock pigeon breeds. MFSD2A, KIRREL3, KCNAB2, and MAPK8IP2 were significantly up-regulated at least 1.3-fold in the homing pigeon. The outcomes of GO enrichment analysis showed that the DEGs in the hippocampus were overrepresented in terms such as cognition, learning or memory and associative learning. In addition, the potential role of GSR in pigeon magnetoreception was reported [Iran is a large country that contains a special range of altitudes and climates. Therefore, there is a remarkable genetic diversity of animal species , 2. In Ireported . The preIt should be noted that, the outcomes of transcriptome data analysis and additional methods details have previously been published in Molecular Biology and Evolution journal . In ordeIn the current study, only four breeds of rock pigeon were investigated and also four tissues were sampled for RNA-seq analysis. In this way, the data set could not present a comprehensive information of brain regions for navigation ability in rock pigeons.