Candida biofilms found on the surface of biomedical devices and in vivo. The action mechanisms of farnesol in fungi have yet to be fully understood, but they are complex and likely include several mechanisms such as growth inhibition and apoptosis promotion.Despite the discoveries of new therapeutic antimycotics, the development of drug resistance is still the main clinical challenge in the treatment of mycoses. Data on the presence of new phytopreparations, along with the direct fungicidal effects that interfere with the resistance of fungal pathogens located in the biofilm, are of great interest. The use of these compounds as monotherapies or in combination with known antimycotics may be an effective strategy for preventing and/or destroying Candida albicans was the first eukaryotic microorganism to exhibit quorum-sensing through the secretion of the sesquiterpene E, farnesol. This molecule is generated by dephosphorylation of farnesyl pyrophosphate in the mevalonate biosynthetic pathway in mammalian and yeast cells. Exogenous farnesol inhibits yeast-to-hyphal formation in a concentration- and time-dependent manner at the earliest stage of hyphal development. Much research has been devoted to studying the role of farnesol as an inhibitor of hyphal morphogenesis; however, little research has been published regarding the in vivo impacts of farnesol on fungal virulence and the development of Candida infection. While other studies have examined the impact of multiple doses of farnesol in addition to antimycotics, we hypothesize that C. albicans treated with a single dose of this quorum-sensing molecule could reduce fungal virulence in a quail model. Candida are unicellular microorganisms, aerobes, of relatively large size and rounded shape, and belong to conditionally pathogenic microorganisms. The genus Candida includes approximately 150 species, which are classified as deuteromycetes due to the complete absence of a sexual stage of development. From a medical point of view, seven of them are recognized as the most important disease-causing species: C. albicans, C. tropicalis, C. krusei, C. kefir, C. glabrata, C. guilliermondii, and C. parapsilosis [Candida are widespread. They live on household items and food, primarily on sugar-rich vegetables and fruits, in dairy products as well as in the internal cavities of mammals including humans [Yeast-like fungi (YLF) of the genus psilosis . In natug humans ,4,5.Candida are capable of acquiring pathogenic properties under certain conditions. In such cases, single or few elements of the fungus (blastospores), which are usually located on healthy mucous membranes, begin to multiply intensively, forming budding cells and multiple filamentous forms (pseudomycelia). The parasitic activity of the pathogen is manifested by the pronounced, rapid formation of a filamentous form. The fungus cells acquiring pathogenic properties attach to the epithelial cells of the mucous membrane, primarily to cells rich in glycogen. The fungi then invade the epithelial cells and parasitize their cytoplasm and nuclei, destroying the host cell [Saprophytic fungi of the genus ost cell ,7.Candida include, first of all, congenital and acquired immunodeficiency conditions; general infections and intoxication; endocrinopathies (diabetes mellitus plays a particularly important role among them); and the violation of the internal environment and the normal microbial “landscape” of the mucous membranes (dysbiosis) under the influence of endogenous and exogenous  influences [In the resulting lesions of the mucous membrane, fungi produce endotoxin and a number of proteolytic, lipolytic, and saccharolytic enzymes. Factors contributing to the manifestation of pathogenic properties, invasiveness, and aggressiveness of fluences ,9.In biological communities, microorganisms use various mechanisms for their communication. Depending on the cell density, bacteria, and YLF can produce QS signaling molecules  that are involved in regulating gene expression and coordinating collective behaviors in their natural niche. The existence of these secondary metabolites plays a major role in the adhesion and colonization of host tissues and surfaces, morphogenesis, and biofilm development . In YLF,C. albicans do not differ significantly from the more general effects of prenylation observed for bacteria, which highlights common mechanisms in biology in both bacteria and fungi.Farnesol is known to chemically modify cysteine residues through a process known as farnesylation, which is a specific sub-example of the more general covalent modification known as prenylation. Alteration of protein prenylation is well known as a factor in the pathology associated with bacterial quorum sensing and biofilm formation, and further mitigation of these effects has shown pharmacological promise ,13,14. AC. albicans incubated with farnesol for one hour and found that this strain can cause a less virulent infection. Our hypothesis is that farnesol could reduce Candida albicans pathogenicity in a quail model.A reasonable solution is to analyze the changes that occur under the action of farnesol on an animal model. In this research, we used Our research focused on four investigations: (1) we analyzed the characteristic aspects of the biofilm development of this YLF strain; (2) we histopathologically analyzed the development of gastrointestinal candidiasis in quails; (3) we compared the hematological blood and serum biochemical analyses; and (4) we observed the changes in microbial composition in quail feces under the action of farnesol. These experiments confirmed related applications of farnesol and its research progression in antifungal therapy.C. albicans collection strain ATCC 10231 was used. For cultivation, we used Sabouraud dextrose agar  and agar with rice extract . Colonies of diurnal cultures of C. albicans from Sabouraud dextrose agar were washed with physiological solution (PhS) (pH 7.0). The concentration of YLF was 0.5, according to McFarland, which corresponded to 1.5 × 108 cells/mL. The optical density (OD) of the biofilm was measured by the degree of binding of crystal violet  at a wavelength of 580 nm (OD580) in an Immunochem-2100 microplate photometric analyzer  [For our experiments, a standardized Farnesol (farnesol) , molar mass = 222.37 gr/mol, mass of the substance = 0.886 g/mL, the amount of the substance in moles = 0.886:222.37 = 0.004 M/mL, or 4000 µM/mL.The daily culture of YLF was washed three times with PhS, and the concentration of YLF was 0.5, according to McFarland. A total of 25 µL of farnesol 100 µM was added to 1 mL of a microbial suspension (experiment), and 25 µL of PhS was added to another test tube (control). Tubes were incubated for 1 h at 37 °C under constant shaking. After interaction with farnesol, YLF was washed three times with PhS ,17.C. albicans processed with farnesol, and the control group was infected with C. albicans without the addition of farnesol.Female, Texas white broiler quails  21 days old, 30 heads, bodyweight 350–370 g, were used. Birds passed veterinary control and had all the documentation a. QuailsC. albicans of 1 mL was given to quails through a digestive probe once a day for five days. Five birds were euthanized every five days. The experiment lasted for 20 days: five days for infection and 15 days for the course of the experiment and observation. The control days for the studies were at five days post-infection, then at 10 and 15 days. In the tables, this is indicated as five days (experience and control), and then 10 and 15 days. Prior to euthanasia, blood samples were taken via the axillary vein in all birds and were examined histopathologically , within 45 min after blood collection, for the total red-blood-cell count (RBC), hemoglobin (HGB), and differential white-blood-cell count/mean leukocytes (LEU) including pseudoeosinophils (PSEUs), eosinophils (EOSs), monocytes (MONOs), basophils (BASs), and lymphocytes (LYMs).The biochemical analysis of blood was performed on the serum within 24 h. Creatinine (CRE), aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), glucose (GLU), mass concentration of cholesterol (CH) and triglycerides (TG), and total bilirubin (TBIL) estimations were carried out using Vetscan VS 2 analyzers .Candida become less aggressive, and were there any changes in the quantitative composition of microorganisms? For this, we prepared serial dilutions of the sample, plated the diluted suspensions, and counted the number of colony-forming units. For YLF, Sabouraud dextrose agar  was used; for Bifidobacterium spp., Blaurock medium ; for Lactobacillus spp., MRS medium ; for Staphylococcus spp., peptone–salt medium, yolk–salt agar ; for Streptococcus spp., mitis-salivarius agar ; for Enterobacteria, Endo’s medium, Ploskirev’s medium, and bismuth-sulfite agar ; for Clostridium spp., clostridial agar ; and for all others, meat-peptone agar . The plates were incubated at 37–38 °C for 24–48 h. To create anaerobic conditions, an AnaeroJar anaerostat was used with Anaerocult gas-generating packages  for 72 h at a temperature of 37 °C. The pure cultures were identified using a matrix-activated laser desorption/ionization technology by MALDI Biotyper . After considering the values of the X score, which ranged from 0 to 3, values from 2 to 3 were considered successful. A result with a score of more than 2.3 was considered to be highly reliable.Samples were taken from the large intestine and placed in sterile test tubes for bacteriological analysis. We were interested in changes in the microbial composition in the large intestine in the experimental and control groups. After treatment with farnesol, would 3 of the milk sample (C) was calculated using the formula and expressed in logarithms with a base of 10:N represents the average number of colonies in 1 bacteriological cup; V represents the volume of suspension, which was applied when seeding the surface of the agar; and K represents the multiplicity of dilution.The number of microorganisms in 1.0 cmp < 0.05.The results were analyzed using SPSS 20.0 . The significance of the results was determined using the Student’s t-test, and the results were considered significant when C. albicans 10231 strain grew abundantly, forming raised, shiny-smooth, white colonies on the Sabouraud agar during 24 h at 28 °C. Under the microscope, the cells were often subspherical with a size of 3 × 4 at 4.5 μm. On day 6 at 28 °C, the culture grown on agar with the rice extract was convex and matte with an uneven edge, white in the center, with a cream color on the periphery. Microscopy found the pseudomycelium branching at an angle of 45° and yeast cells with a diameter of 3.6 μm. Terminal chlamydospores had formed.The C. albicans 10231 in the control group (0.497 ± 0.05) showed that this microorganism produced a significant biofilm  of  biofilm . The YLFCandida fungus were detected. After being euthanized on day 15, the remaining four quails were found to have typical, well-marked candidiasis lesions in the anterior part of the digestive tract. In the histological sections, multiple filaments of the fungus were found growing perpendicular to the thickness of the mucous membrane.In the control group of birds euthanized within the first five days post-infection, swelling, weak hyperemia of the goiter mucosa, thick viscous mucus, and a delicate whitish plaque containing an abundance of budding cells and pseudomycelia were observed. In the histological sections of the goiter, oral cavity, and esophagus, fungal ingrowth into the epithelial cover of the mucous membrane had begun. On day 10 post-infection, the curd overlays were more distinct, with rounded foci. On day 15, we observed clinical signs of the disease in the form of depression, drowsiness, and poor appetite. One of the five remaining quail died prior to euthanasia. Upon autopsy, the mucous membrane of the goiter appeared “bumpier” due to the different intensity of overlays, which in sections took the form of a solid yellow-white film, and the serous membrane had folded. Delicate, loose, yellow-white overlays were noted on the mucous membrane of the oral cavity and tongue, in which multiple budding and pseudomycelial forms of the Candida albicans were found.In the experimental quails, where the YLF was treated with farnesol, we did not observe any hyperemia of the goiter mucosa in the first five days post-infection. In the histological sections of the goiter, oral cavity, and esophagus, fungal ingrowth into the epithelial cover of the mucous membrane was detected, but to a lesser extent than in the controls. On day 10 post-infection, we observed a delicate whitish plaque on the mucous membranes of the digestive tract, containing a small number of budding cells. On day 15 pre-euthanasia, clinical signs of the disease in the form of depression, drowsiness, and poor appetite were not observed in the birds, and they had all survived to this point. Upon autopsy, the mucous membrane of the goiter was found to be covered with a delicate white biofilm, in which budding C. albicans had progressed, either as a biofilm (controls) or individual cells not connected by a matrix . In previous experiments where Candida was in the form of a biofilm, farnesol had been effective. However, microorganisms can also form biofilms in living models and, therefore, farnesol needs to be further tested. Our results suggest that farnesol reduced the pathogenicity of Candida and slowed the progression of the infection in living animals. Moreover, the YLF were less capable of forming a biofilm while in the presence of farnesol.Our results indicated that there was a significant difference in the form to which The results of the hematological and biochemical analyses in the experimental and control groups are summarized in Candida.Pronounced violations of the microbial spectrum in the large intestine were found, which were identified as dysbiosis caused by a facultative decrease in the indigenous microflora of the large intestine and an increase in conditionally pathogenic levels of Candida in the control group did not decrease. In the experimental group, where the Candida was treated with farnesol prior to infecting the quails, the amount of YLF decreased. This was due to the decreased ability of the fungi to form biofilms after treatment with farnesol. A significant difference was found in the amount of normoflora in the experimental vs. control groups. In the experimental group, Lactobacillus spp. and Bifidobacterium spp. showed better resilience due to not being hindered by pathogenic Candida.As shown in The differences between the experimental and control groups were clearly visible when we recalculated our results as percentages. We assigned all microorganisms in this group at 100%, and then the shifts in microorganisms in the presence of farnesol was immediately visible, as shown in Candida spp. has forced scientists worldwide to consider new approaches for antimycotic drugs. A prerequisite is the safety and non-toxicity of new drugs for the patient [C. albicans is its ability to switch from yeast to hyphae since the hyphal form can adhere and penetrate tissues more readily than the yeast form. The first stage in this transition is the formation of a germ tube, which is triggered by interaction with the host cells and is dependent on factors such as serum, pH, temperature, and quorum-sensing molecules (QSMs) [The increasing dominance of polyresistant strains of  patient ,6,21. Ons (QSMs) ,23.C. albicans via the accumulation of reactive oxygen species (ROS), mitochondrial degradation, and caspase activation [The first identified QS molecule, farnesol, attracted interest, and while significant progress was made ,24, the tivation .Candida biofilm as well as the concentration used. However, to date, the effect of farnesol and its synergy and antagonism with known antifungal drugs has not been comprehensively studied. In our experiment, we found that farnesol affected the degree of YLF resistance, which supports its potential as an antimicrobial agent [Furthermore, farnesol and its derivatives have exhibited antibiofilm, anticancer, antitumor, and fungicidal properties ,21,29. Tal agent ,30,31.Candida infection. While other studies have examined the impact of multiple doses of farnesol in addition to antimycotics [C. albicans treated with a single dose of this quorum-sensing molecule could reduce fungal virulence in a quail model.Much research has been devoted to studying the role of farnesol as an inhibitor of hyphal morphogenesis ,32; howemycotics , we hypoC. albicans cells sense farnesol or how this QS molecule exerts its biological effects remains unclear [In our previous studies, we have shown that farnesol affected the formation of biofilms to a greater extent than planktonic cells. However, despite intensive research on farnesol over the last decade, how  unclear ,35.Lactobacillus spp. and Bifidobacterium spp. were present in greater concentrations when Candida was treated with farnesol. Given the increased patient sensitivities to existing medications reported in the literature, reducing pathogenicity and reducing biofilm formation via farnesol could be significant for the development of future therapies. This experiment may also provide a deeper understanding of the potential antifungal mechanism of QS molecules as we search for new solutions to counter infections involving Candida. Combined with conventional antimicrobial therapies, the therapeutic potential of this QS molecule on the virulence factors of pathogens such as C. albicans should be considered for biofilm-associated diseases.In summary, our results suggested that the candidiasis of the digestive tract developed to a lesser extent if treated with farnesol, and farnesol had a significant effect on the microbiological composition of the intestines.


Background: The existing prediction models lack the generalized applicability for chronic heart failure (CHF) readmission. We aimed to develop and validate a widely applicable nomogram for the prediction of 180-day readmission to the patients.Methods: We prospectively enrolled 2,980 consecutive patients with CHF from two hospitals. A nomogram was created to predict 180-day readmission based on the selected variables. The patients were divided into three datasets for development, internal validation, and external validation . At baseline, 102 variables were submitted to the least absolute shrinkage and selection operator (Lasso) regression algorithm for variable selection. The selected variables were processed by the multivariable Cox proportional hazards regression modeling combined with univariate analysis and stepwise regression. The model was evaluated by the concordance index (C-index) and calibration plot. Finally, the nomogram was provided to visualize the results. The improvement in the regression model was calculated by the net reclassification index (NRI) .Results: Among the selected 2,980 patients, 1,696 (56.9%) were readmitted within 180 days, and 1,502 (50.4%) were men. A nomogram was established by the results of Lasso regression, univariate analysis, stepwise regression and multivariate Cox regression, as well as variables with clinical significance. The values of the C-index were 0.75 [95% confidence interval (CI): 0.72–0.79], 0.75 [95% CI: 0.69–0.81], and 0.73 [95% CI: 0.64–0.83] for the development, internal validation, and external validation datasets, respectively. Calibration plots were provided for both the internal and external validation sets. Five variables including history of acute heart failure, emergency department visit, age, blood urea nitrogen level, and beta blocker usage were considered in the final prediction model. When adding variables involving hospital discharge way, alcohol taken and left bundle branch block, the calculated values of NRI demonstrated no significant improvements.Conclusions: A nomogram for the prediction of 180-day readmission of patients with CHF was developed and validated based on five variables. The proposed methodology can improve the accurate prediction of patient readmission and have the wide applications for CHF. It is well-known that chronic heart failure (CHF) is a systemic clinical syndrome as well as an endpoint stage of various cardiovascular diseases with typical symptoms of systolic and/or diastolic dysfunction. In recent years, existing treatments are effective in reversing the progression to end-stage disease among patients diagnosed with CHF; however, the mortality and readmission rates are still high . To bettIn developed countries, some physicians in the United States focus on the 30-day readmission rate , thus leThe flowchart of the selection of patients for the multicenter prospective cohort study is depicted in We selected patients hospitalized for CHF between January 2019 and June 2019 at two centers, where one was a North China center of HF  and the other was a South China center of HF . The patients would be enrolled if they met the following criteria: patients with CHF and diagnosed with various etiologies, New York Heart Association (NYHA) class II–IV, and age >18 years. Moreover, the exclusion criteria were as follows: planned HF readmission within 6 months , missing data for the essential variables and outcomes (patients missing 30% or more variables will be excluded), severe patients with liver and kidney dysfunction and/or malignant tumors, and poorer treatment compliance . Severe liver dysfunction is defined as an alanine aminotransferase level more than twice that of normal or Budd–Chiari grade C. Severe renal dysfunction is defined as chronic kidney disease stage V. As per the above indicators, the definitions are provided in further studies.It should be stressed that this study was approved by the ethics committee  of each chosen hospital and was also conducted in accordance with the Declaration of Helsinki (ChiCTR1800019869). Furthermore, the participants from both hospitals provided informed written consent for supporting the clinical research studies.In this study, the main predictors include sociodemographics, clinical characteristics, comorbidities, results of laboratory tests (blood sample assay), other relevant physical examinations, medication usages in the hospital and at discharge, and other treatment measures. In detail, the sociodemographics contain basic information, such as age, sex, the distance between the hospital and residence, length of hospital stay, medical social insurance status, and other variables associated with readmission. In addition, we collected some samples in the progress of treatments, as detailed as possible, and selected the features that corresponded to the prognosis of the disease. Among these predictors, comorbidities, complications, and Charlson comorbidity index (CCI)  were recThe diagnosis of CHF is based on the 2016 ESC Guideline for HF management, and patients were diagnosed by two experienced cardiologists in each HF center . In the k = 0.7  or 0.9 , α = -0.329 or-0.411 , Cr = plasma creatininewhere1 mg/dl = 88.4 μmol/l .The prediction model for CHF can be established according to the statements in the transparent reporting of a multivariable prediction model for individual prognosis (TRIPOD) guidelines , where wIn addition to the internal validation cohorts , an external validation cohort (comprising 293 patients) was developed from a prospective study at the First Affiliated Hospital of Medical College in Shantou University. Based on the development and validation cohorts, numerical comparisons will be provided by the proposed methodology.The primary outcome is unplanned HF readmission within 180 days after the first admission for the referred patients. They are collected from both centers mentioned above, and they are followed up for 180 days after discharge from the index admissions. To better track the patients, we adopted the way by telephoning the interviews and further verified their conditions by virtue of the hospital system.We provide the numerical comparisons for the continuous variables, which are described as the mean ± standard deviation. Meanwhile, categorical variables are described as percentages or frequencies. Continuous variables are divided into categorical variables based on the results of restricted cubic spline according to data distribution and its clinical implications and boundaries . Hazard www.cran.r-project.org/version 3.6.1) on a personal computer.In the numerical verification, both internal and external comparisons are conducted for the desired expectations. The concordance index (C-index), with 200 bootstrap samples, and the calibration plot are considered to evaluate the prediction accuracy as well as test the consistency of the prediction model, respectively. Besides this, the NRI is provided to compare the different models in the numerical experiments. All of the statistical analyses are conducted using SPSS version 25.0  and the R Project package for statistical computing  and cross-validation (B) results].The model is developed from 102 variables by screening with the Lasso–Cox regression algorithm. The most prominent advantage of Lasso regression is that it penalizes the coefficients of all variables by regression and makes the relatively unimportant independent variable coefficients zero by the regularization technique. Therefore, when we use Lasso regression, we have retained the punitive and inhibited the collinearity. Results of Lasso regression are shown in p > 0.05): the p-values for AHF, BUN, emergency, eGFR, and NT-proBNP were 0.77, 0.27, 0.38, 0.79, and 0.88, respectively. They were identified as independently associated with the 180-day HF readmission. The results of collinearity diagnosis of the five selected variables are shown in the The five selected variables consisted of AHF, emergency department visit (emergency), eGFR, N-terminal pro-B-type natriuretic peptide (NT-proBNP) and blood urea nitrogen (BUN) level. It supported the proportional risk hypothesis (r = 0.137), β-blocker usage (r = −0.141), hospital discharge way (r = 0.136), alcohol taken (r = 0.131), and left bundle branch block (r = 0.097).At the same time, we combined the results with univariate Cox analysis and stepwise regression to avoid missing variables that were clinically significant. According to the results of univariate analysis and stepwise regression, 24 variables were selected from 102 baseline variables, and 24 variables were allocated to five groups (the group was divided into five groups according to the Lasso–Cox result) according to the model inclusion principle of clinical significance . In eachNRI  was assep < 0.01), 3.342 , 1.201 , 0.722 , and 1.132 , respectively (depicted in p < 0.001).Considering the Akaike information criterion  and convhttps://youyueer.shinyapps.io/dynnomapp/).A nomogram is built to visualize the risk of readmission by using the selected predictors according to the probability values obtained by Cox regression modeling in the development set . FirstlyThe validation of the prediction model in the internal validation set is evaluated by discrimination and calibration. A calibration plot further shows good fitness with the actual readmission rates . It should be specially noted that the calibration plot can show good consistency in predicting the 180-day readmission risk for patients with CHF , indicating that the nomogram could correctly predict individuals with a 180-day unplanned readmission in the external validation set.For patients with CHF, an accurate prediction of the readmission rate will aid in clinical stratification and treatment decision-making for the development of appropriate diagnosis and treatment programs and the development of healthcare and management guidelines for HF. In this study, we established an easy-to-use nomogram based on easy-to-get clinical variables and validated in patients in a second hospital . This ensured wide applicability of the prediction model. This is the first 180-day CHF readmission model based on data of hospitals in North and South China. Compared with previous HF readmission models , we consA 90-day readmission model of HF is developed by Tan et al. from the South China. The C-statistic of the model of Tan is 0.732. The model shows a moderate predictive accuracy; however, 95% CI and validation are not given. Another HF model is developed by Yang et al. and they estimate 30-day  and 1-year readmission rates . The model is developed in North China and lacks external validation. Our nomogram shows a similar accuracy  and we validate it in internal and external sets. However, more mental variables  and social support should be taken into account in further studies. Han's model  enrolls mental variables, but the common variables of HF and the lack of internal and external validation are neglected. Hughes' analysis of 30- and 180-day readmission showed a lower accuracy .In terms of validation, our model is validated by cross-validation, internal validation, and external validation. We used 10-fold cross-validation , that isIn terms of applicability, we used two CHF prospective study cohorts from HF centers in both North and South China, an optimal prognostic prediction model for 180-day readmission, including age, AHF, β-blocker, BUN, and emergency. Different data sources ensure the general applicability of the model. Besides, Lasso regression made the coefficients of some useless variable features penalized and even made some coefficients with smaller absolute values directly forced to zero. Finally, the penalty condition was satisfied and the sum of residual squares was minimized so as to enhance the generalization ability of the model.Lasso regression used the following formula :p refers to the number of the variable; n refers to the number of samples. Satisfactory consistency with good calibration was observed in the independent external validation set. The developed nomogram robustly quantified the risk of readmission of an individual within 180 days of discharge. The intuitive features easily allowed the clinical staffs to predict the readmission and prognosis of patients with CHF using several important symptoms and clinical indicators.Penalty condition: Prediction variables independently associated with readmission are included in our model. Two predictors (AHF and emergency) are associated with the medical history of the patients. AHF  is commoThe emergency treatments are important indicators in the 180-day readmission model for CHF. The emergency department is the key point of initial treatment for patients with AHF attack. The emergency department plays an important role in the management and treatment of AHF patients . TherefoWith the aging process, the number of CHF patients is increasing. In developed countries, the incidence rate of HF is about 1–2% in adults, while in elderly people over 70, the figure rises to nearly 10% . The preIn the present study, we mainly consider plasma BUN level. The level of BUN reflects the protein metabolism, indicating the renal function, which is independently associated with the prognosis of patients with CHF. Moreover, they are also selected for model inclusion. Fluid overload is a common pathophysiological mechanism of CHF as well as renal disorders. There exist complex neurohormonal interactions between the heart and kidney for patients with CHF. Patients with CHF have a higher incidence of renal dysfunction due to several shared pathophysiological pathways and mutual risk factors . The exiIn order to design new strategies to improve clinical outcomes, a better understanding of the mechanisms has to be achieved, and the experimental results for the hemodynamic changes affecting the lungs and kidneys in HF need to be validated further.eGFR is another factor to be analyzed; however, it is a sensitive factor together with the change of blood creatinine. Improvement is not obvious when we add eGFR to our prediction model. This is similar to the conclusion of an HFpEF study . This maFrom the analysis, we observed the benefits of β-blocker usage in CHF patients. In future studies, we would consider the dosage of β-blockers in detail, as well as the individual differences when choosing the requirements of drug use. The elderly aged ≥75 years who received a β-blocker hold a lower 90-day mortality rate and lower readmission rate . ConsequFinally, the five variables associated with CHF readmission entered into the model were as follows: AHF, emergency, age, BUN, and β-blocker usage. This fits with the AIC principle. When the predictive effect is similar, we tend to choose fewer variables for clinical applicability.Several studies focused on the enhancement of the accuracy of the models  but ignoThis work mainly provides a simple model, which can address data collected for clinical practice to predict the 180-day readmission of patients with CHF. Both internal and external validations further suggest a broad applicability of the given model. From the results, the prediction model can show a reference value for a meaningful prognosis to stratify patients with CHF and then aid in timely clinical decision-making.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Qingdao Central Hospital Ethics Committee, The Ethics Committee of the First Affiliated Hospital of Shantou University Medical College. The patients/participants provided their written informed consent to participate in this study.SG: conception and methodology, project administration, and writing of the original draft. GY: conception and methodology and data curation (from the North China Heart Failure Center). QX: conception and methodology and data curation (from the North China Heart Failure Center). GW: conception and methodology and data curation (from the South China Heart Failure Center). XT: conception and methodology and data curation (from the South China Heart Failure Center). JZ: writing, review, and editing. NL: writing, review, and editing. All authors contributed to the article and approved the submitted version.This study was supported by the Guangdong High-Level University Development Program Research Fund (2020) and Innovation Team Project of Guangdong Universities, China .The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


In recent years, pigs involved in the dry-cured ham production system have suffered from excessive leanness. This has led to the increase of slaughter weight (SW) to achieve greater carcass and ham fatness statuses to compensate for the loss in dry-curing aptitude. The production guidelines for the production of Italian dry-cured ham are currently under revision and an extension of the range of carcass weights from 126 to 168 kg, corresponding to about 146 to 210 kg of SW, has been proposed. However, little is known about the influence of SW in the range of 140–200 kg on growth performance, feed efficiency, carcass quality and ham curing aptitude. We hypothesized that an increased SW could exert a positive influence on ham characteristics. Data from 159 pigs fed ad libitum with diets, unlimiting for nutrient contents, up to 8 or 9 months of age (140–200 kg SW) were used. Greater SWs were linearly and positively associated with the growth performance of the pigs and with better ham quality traits. Greater SW increased ham weight, muscularity, and greater fat covering, according to the dry-cured ham industry’s expectations. Barrows produced hams with greater weight and marbling than gilts.semimembranosus muscle. Barrows were heavier and produced hams with slightly better characteristics than gilts.Slaughter weight (SW) is critical for dry-cured ham production systems with heavy pigs. A total of 159 C21 Goland pigs (gilts and barrows) at 95 ± 9.0 kg body weight (BW) from three batches were used to investigate the impact of ad libitum feeding on SW, growth performance, feed efficiency, and carcass and green ham characteristics. Diets contained 10 MJ/kg of net energy and 7.4 and 6.0 g/kg of SID-lysine. Slaughter weight classes (SWC) included <165, 165–180, 180–110 and >210 kg BW. In each batch, pigs were sacrificed at 230 or 258 d of age. Left hams were scored for round shape, fat cover thickness, marbling, lean colour, bicolour and veining. Data were analyzed with a model considering SWC, sex and SWC × Sex interactions as fixed factors and the batch as a random factor. The linear, quadratic and cubic effects of SWC were tested, but only linear effects were found. Results showed that pigs with greater SWC had greater average daily gain and feed consumption, with similar feed efficiency and better ham quality traits: greater ham weight, muscularity, and fat coveringin correspondence of  The major limitation for increases in pig slaughter weights (SWs) is increases in carcass adiposity and the worsening of feed efficiency with increasing SW . HoweverFor dry-cured ham production, adequate fat covering and marbling are required, so the pigs must be slaughtered at heavy weights, often greater than 130 kg . In thesThe extension of the admitted SW range implies the possibility of adopting an ad libitum feeding strategy that would better exploit the genetic potential of individual pigs for growth—although with a reduction of body and carcass uniformity among pigs of the same batch ,10. TherAssuming that pigs selected for dry-cured ham production are slaughtered at about the same age, it can be hypothesized that those heavier at slaughter would be those with greater feed consumption, growth rate and carcass and ham weight, but also with greater carcass adiposity, ham marbling and ham fat covering. In addition, some loss in feed efficiency may occur, increasing the SW . HoweverThis paper aimed to study, in groups of pigs fed ad libitum and selected for dry-cured ham production, the relationships between SW, growth performance, feed efficiency, and carcass and green ham characteristics.The data used in this research originated from a previous experiment that involved 336 pigs, from three batches of 112 pigs each . BrieflyPigs were members of 68 full-sib families of the C21 Goland boar line , generated by mating 13 boars to 67 sows. Besides growth and residual feed efficiency, the breeding goal of the C21 Goland line includes traits related to the quality of raw ham  and its In early (90 to 120 kg BW) and late (120 kg BW upwards) finishing periods, the pigs received cereal–soybean meal-based diets . The feeAt the start of the experiment, and the day before slaughtering, the pigs were weighed with a scale. The pigs of the YA and GW groups were reared in the same way, but they were slaughtered at different ages. These two groups had homoscedastic variances and ample variations in SW.biceps femoris (P1) and semimembranosus (P2) using a ruler or a portable ultrasound system , respectively.The pigs of the YA and GW groups were slaughtered, on average, after 85 or 116 days on feed—corresponding to almost 8 or 9 months of age. An extra month of feeding would increase the SW, the daily and cumulated feed consumption, the carcass and the ham fat covering, the ham size, and would reduce the average daily gain and feed efficiency. Slaughter and carcass dressing were carried out as described in . Hot carHams were moved to the ham factory within two days after the slaughter, where they were trimmed again and weighted. The hams were trimmed to obtain the typical shape of Veneto ham, without the trotter.According to current guidelines, 160 kg ± 10% is the average weight of the batch. Accordingly, data were grouped into four SW classes (SWC), with about 20 kg SW of difference between one class and the following one. The first SWC represented pigs with lighter SW (<165 kg SW), which is still in agreement with current guidelines. The second SWC (165−180 kg SW) were somewhat heavier pigs, with SWs similar to those frequently found in practice. The third SWC represented pigs with SWs  in agreement with the proposal of the guideline revisions, and the fourth SWC (>210 kg) represented pigs that were too heavy and would be discarded if the new production guidelines proposal are approved. ijklm was the observed trait, µ was the overall intercept of the model, SWC was the fixed effect of the thi class of SW , Sex was the fixed effect of the thj sex , (SWC × sex) was the interaction effect between the SWC class and sex, Batch was the random effect of the thk batch , and eijkl was the random residual. Carcass and ham trait data were analyzed using a GLM procedure in SAS  using the following linear model:2l and σ2e, respectively. SWC, Sex, and SWC × sex interaction effects were tested in relation to the residual variance . Three degrees of freedom of SWC were used to test the linear, quadratic and cubic effects of increasing SWC. As the quadratic and the cubic components were never significant, the p-values of these components were omitted from the tables.The Batch and the residuals were assumed to be independently and normally distributed, with a mean of zero and a variance of σb) relating carcass weight to SW, and lean and fat masses to carcass weight were fitted using a spreadsheet.Allometric relationships (y = axp < 0.001), but feed efficiency (gain: feed) did not (p = 0.53). As expected, the lighter SWC were represented in greater proportion by YA pigs, and the heavier SWC by the GW pigs. The most frequent class was the third, followed by the second, the first and the fourth . Pigs wip < 0.001), carcass yield (p < 0.001) and carcass backfat depth (p < 0.001) linearly increased, whereas the lean meat percentage decreased (p < 0.001). The allometric coefficient relating carcass weight to SW was greater than 1.00 (1.046), as the increase in SW was associated with a more than proportional increase in carcass weight  and fat cuts (p < 0.001) increased with increasing SWC, while the carcass yield of lean cuts decreased (p < 0.001), and that of fat cuts increased (p < 0.001). The relationships of the lean and the fat cuts on carcass weights evidenced allometric coefficients lower than one (b = 0.855) and greater than one (b = 1.342), respectively (The weights of lean (ectively .p = 0.018). However, the barrows had greater SW (p = 0.027), carcass yield (p = 0.039), fat cuts weight (p = 0.043), and lower lean cuts yield (p = 0.026). The Sex × SW interaction had negligible influence on growth performance and major carcass traits.Sex had little influence on feed intake, average daily gain, SW, carcass weight, carcass yield, carcass backfat depth and total and lean cut weight. The feed efficiency of the barrows was somewhat lower than that of the gilts (p < 0.001) with increasing SWC (p < 0001), and those of the fat cuts, back fat and belly increased (p < 0.001). The trimming losses increased with increased SWC (p < 0.001), both in terms of weight and yield. The barrows had greater belly weight (p < 0.001) and yields (p = 0.006) than gilts, but lower yields of loins and ribs (p = 0.003).All the various wholesale cuts weights increased linearly (sing SWC . Howeverp < 0.001), at the ham factory (p < 0.001), and the trimming losses at the ham factory (p < 0.001) linearly increased with increased SWC (p = 0.002), and a greater subcutaneous fat depth in the P2 position (p = 0.005). However, the SWC class had little influence on other ham quality parameters.The weights of trimmed ham at the slaughterhouse (ased SWC . Pigs wip = 0.037) and greater marbling (p = 0.011) than gilts. The Sex × SWC interaction had no influence on these ham traits.Sex also had little influence on these ham characteristics, except for marbling and hemorrhage. Barrows had a lower hemorrhage score  and 258 d (8.6 months) of age, and the SW ranged from a minimum of 137 to a maximum of 225 kg. With increasing SWC, the frequency of pigs slaughtered at younger ages decreased, and that of pigs slaughtered at older ages increased. This partial confusion between age and SW was accepted, as it may become representative of the commercial conditions in the case of application of the innovative rearing strategies proposed in Malgwi et al [The results of the current experiment can be compared with others ,31 achieThe first relevant finding of the current paper is that feed efficiency was not related to the increase in SW. This is in apparent contradiction with the literature, which reports that feed efficiency decreases with increasing physiological maturity . These aThe pigs in the current research evidenced good potential for growth at heavy BWs, both for lean and fatty tissues. Besides growth and residual feed efficiency, the breeding goal of this line includes traits related to the quality of raw hams  and theiThe proposal of new guidelines for dry-cured ham production indicates that carcass weights must range between a minimum of 120 and a maximum of 168 kg. In our research, 10 out of 159 carcasses (6.2%) were heavier than the upper limit. This would suggest that, when fed ad libitum, some Goland C21 pigs would be heavier than the maximum indicated for dry-cured ham production. As an anticipation of the age at slaughter might be not permitted by the guidelines, this shortcoming would be resolved by introducing a mild feed restriction or practices of precision feeding, resulting in benefits in terms of uniformity  of the pIn the current experiment, the coefficient of variation for carcass weight was 11%, whereas in previous research, where pigs were kept on restricted feeding regimes, the coefficient of variation was in the order of 6−7% . Herein,p < 0.01). Such an increase in carcass yield is due to the differential development of the carcass fatty and lean tissues compared to the non-carcass parts [p < 0.01). The correlation between carcass weight and carcass yield was appreciable and positive  lighter than the minimum required to achieve the label. The weight of the trimmed ham was further reduced according to the additional trimming procedure conducted at the local ham factory.The ham weight and size, together with the inter-and intramuscular fat content, the thickness of the subcutaneous fat and the lean meat content of the hind leg, represent the main factors that can also influence the aptitude of the ham to adsorb salt . It is csemimembranosus muscle, and on the roundness—a measure of muscularity. Interestingly, with increasing SW, the subcutaneous depth of the carcass increased, but the subcutaneous fat depth of the ham increased only in correspondence to the semimembranosus muscle, and not in correspondence to the biceps femoris. This seems to not be fully consistent with the results of [biceps femoris muscle (r = 0.53), rather than to the semimembranosus muscle (r = 0.18). In the current experiment, there was no correlation between the average carcass fat thickness and the measures of subcutaneous fat thickness taken at the two positions, with simple correlation coefficients ranging from 0.06 to 0.02. However, in agreement with previous research [semimembranosus muscle was much thinner than that measured in correspondence to the quadriceps femoris muscle. As the thickness of subcutaneous fat influences salt penetration and water seasoning losses during seasoning [semimembranosus muscle may exert a critical role in determining the dry-curing aptitudes of ham [biceps femoris muscle, which may not be desired by the consumer and may limit the marketability of the ham [In the current experiment, the increased SW had little influence on the majority of the ham’s quality traits, except on the subcutaneous fat depth, corresponding to the sults of , who fouresearch ,39, the easoning , measures of ham . This re the ham . The influence of a thicker round shape on the seasoning aptitude of the hams, or globosity, is poorly described in the literature. In practice, a greater ham roundness is frequently associated with excessive leanness, scarce subcutaneous fat covering, greater water content and salt absorption, greater seasoning losses and poor final quality of the seasoned ham . In the In the Italian heavy pig production system, previous research has found little differences between gilts and barrows . Such a Some differences between barrows and gilts were also found for some subjective scores, as barrows scored lower for hemorrhages and greater for visible marbling compared to gilts. However, the magnitude of these differences was modest. Therefore, it appears that no solid reasons can be given, at this point in time, to indicate that barrows are better than gilts when intended for Italian dry-cured ham production. This is dependent on the pig genetic line, as in other productive contexts, barrows have been found to be better than gilts when intended for dry-cured ham production .semimembranosus muscle. Greater ham weight and fat covering in correspondence to the semimembranosus muscle are desired by the dry-cured ham industry for better curing aptitudes. Barrows produced hams with greater weight and marbling than gilts. A greater marbling is desired because of its positive influence on the flavor and the visual traits of green ham at the time of its selection for dry-curing. These characteristics are evaluated by the dry-cured ham industry before the curing process for better profitability and consumer acceptability of the seasoned product. Data from this research also indicate that pigs of the Goland C21 genotype can reach the traditional weight of 160 ± 16 kg at only 8 months of age—one month less than the traditional age. New knowledge about the influence of slaughter age on the seasoning aptitude of the hams, not confounding SW with slaughter age, is desired.In conclusion, pigs with greater SWCs had greater average daily gain and feed consumption with similar feed efficiency, greater ham weight, muscularity and fat covering in correspondence to the


Minor hallucinations (MHs) are the most common psychotic phenomena in Parkinson's disease (PD), and it has important clinical and prognostic implications in PD. Plasma homocysteine (Hcy) has been reported to predict the outcome of PD; whether or not Hcy is associated with MH is not known. We aim to investigate the Hcy level and related factors in patients with PD and MH.  We conducted a cross-sectional study and included 99 patients with PD, 34 with MH, and 65 without any hallucinations. The clinical and demographic data of the patients with and without hallucinations were compared. Hcy-related clinical factors were also analyzed.  The plasma Hcy level was higher in MH patients than in patients without hallucinations, and the result was more pronounced in male patients than in female patients. Differences were also observed when the groups were divided on the basis of levodopa equivalent daily dose and disease duration. The high Hcy concentration was correlated with some symptoms in patients with MH, including motor dysfunction and nonmotor symptoms, such as symptoms of the gastrointestinal tract, angiocarpy, sleep/fatigue, and poor visuospatial/executive function.  Results indicated a higher plasma Hcy concentration in MH patients than in their counterparts and revealed that Hcy is associated with certain motor and nonmotor symptoms in patients with MH. Hcy may be a marker of MH and have important therapeutic implications in PD. Parkinson's disease (PD) is the second-largest neurodegenerative disease with clinical manifestations of motor and nonmotor symptoms. As a common nonmotor symptom, minor hallucinations (MHs) have been recognized as a premotor symptom , 2 and tHomocysteine (Hcy) is an important intermediate product produced during the metabolism of methionine, known as the methionine cycle. As shown in High-level Hcy is closely related to vascular diseases; evidence has shown that it also plays an important role in neurodegenerative disorders, such as PD . PatientMotivated by our previous observation of the plasma Hcy levels of patients with PD presenting MHs, we aim to investigate the Hcy level and related factors in patients with PD and MH.n = 34). Those with a score of 1 and fulfilled the condition in which MHs occurred steadily and more than once per week in the last three months before the study were enrolled in the PD-MH group (n = 65). A questionnaire [From May 2019 to January 2021, 210 patients with idiopathic PD were consecutively recruited from the Affiliated Brain Hospital of Nanjing Medical University, Nanjing, P. R. China. The diagnosis of PD patients was carried out by at least two movement-disorder specialists on the basis of the PD United Kingdom Brain Bank criteria . The excionnaire  of 12 itionnaire , 16, we ionnaire , and disThis study was approved by the local Ethics Committee of the Brain Hospital Affiliated to Nanjing Medical University and was conducted in accordance with the principles outlined in the Declaration of Helsinki. All participants signed informed written consent and provided clinical data.The following demographic information was collected using a standard questionnaire: age, gender, marriage, education level, body mass index (BMI), history of smoking and drinking, history of drinking tea and coffee, daily exercise, family history of PD, predominance of motor symptoms, disease duration, and use of antiparkinsonian drugs .Motor disability and disease severity were assessed using the Unified Parkinson's Disease Rating Scale (UPDRS) part III score and the Hoehn and Yahr (H–Y) scale, respectively. H–Y stage ≤ 2 was defined as early stage while H–Y stage > 2 as the advanced stage. The Non-Motor Symptoms Questionnaire (NMS-Quest) was used to measure the nonmotor symptoms in PD . Global μmol/L, nmol/L, and pmol/L, respectively.After fasting overnight, 3 mL of venous blood was collected from all patients at 7 am. The blood samples were placed on ice and then centrifuged at 3,000 rpm for 10 min to obtain the plasma. The levels of Hcy, folate, and VB12 were measured from plasma. These steps were completed within 2 h. Hcy was determined by enzymatic cycling technology using an Hcy reagent kit  in an automatic biochemical analyzer . Enzymatic cycling technology uses enzymes to convert Hcy into a free form, reacts with covalent substrates, and circulates amplification. The final product converts nicotinamide adenine dinucleotide (NADH) into nicotinamide adenine dinucleotide. The concentration of Hcy is detected by measuring the absorbance of NADH. Chemiluminescence microparticle immunoassay technology, which combines magnetic separation technology, chemiluminescence technology, and immunoassay technology, was conducted using the ARCHITECT i system  of a folate reagent kit  and a B12 reagent kit  to investigate the plasma levels of folate and VB12, respectively. The levels of Hcy, folate, and VB12 were expressed in t-test was used; the Mann–Whitney U test was conducted otherwise. Categorical variables were shown as percentages and analyzed using a chi-square test and Pearson's and Fisher's exact test. When the data were ordinal, the Mann–Whitney U test was used. Correlations between the clinical characteristics and Hcy levels were performed using Spearman's rank correlation coefficient (rs). SPSS 23.0  was used in the data analysis, and GraphPad Prism 8.0 was used to draw histograms. Values of P < 0.05 were deemed statistically significant.Our research is mainly a comparison between two groups. In this study, all continuous variables were presented as the mean ± standard deviation (SD). The Kolmogorov–Smirnov test was used to analyze the normal distribution of all variables. When the data conformed to the normal distribution, Student's P = 0.001), and the subdomains of the NMS-Quest scale, including gastrointestinal tract, depression/anxiety/anhedonia, sleep/fatigue, and miscellaneous , were significantly different between them . Among all the nonmotor symptoms of PD, the problems of sleep quality (P = 0.028) and pRBD (P = 0.04) were more severe in the patients with MHs than in those without. Compared with the patients without any hallucinations, those with MHs had lower scores in PDQ39 (P = 0.028) and relatively higher scores in HAMA (P = 0.084). No significant differences were noted in the scores of HAMD (P = 0.279) and MoCA (P = 0.84); however, the scores of MoCA in the visuospatial/executive domain were relatively high in the nonhallucinators (P = 0.034). In this study, the plasma level of Hcy was higher in the PD-MH patients than in the PD-NH patients (P = 0.001). Meanwhile, the plasma level of folate tended to be higher in the PD-NH group than in the PD-MH group (P = 0.053) while no significant differences in VB12 levels were noted between the minor hallucinators and the nonhallucinators.The clinical characteristics of all the patients are summarized in P = 0.012). Meanwhile, the Hcy levels exhibited an upward trend in the female patients in the PD-MH group relative to those in the PD-NH group (P = 0.05). Folate levels displayed significant differences between the male and female patients . VB12 levels did not present any differences in the subgroups . When the patients were further divided into three subgroups on the basis of LEDD, we found that plasma levels were significantly higher in the PD-MH patients than in the PD-NH patients in the LEDD > 750 subgroup (P = 0.043). The subgroup of 375 ≤ LEDD ≤ 750 was tended to have a high level of Hcy (P = 0.078). When disease duration > 5 years, the PD-MH patients had a higher level of Hcy than the PD-NH patients (P = 0.002).When the patients were divided into specific groups according to LEDD, our data  showed nP = 0.04, 0.014, and 0.036, respectively) and positively correlated with UPDRS III; NMS total score; and NMS gastrointestinal tract, cardiovascular, and sleep/fatigue subdomains . In the PD-MH patients, no significant correlations were noted between Hcy and age, Hcy and LEDD, Hcy and NMS other subdomains, Hcy and PDSS, Hcy and total MoCA, Hcy and HAMA, Hcy and HAMD, and Hcy and PDQ39.The results of Spearman's correlation analysis among Hcy and the other clinical assessment scales for the PD-MH patients are summarized in In this cross-sectional study, we found that apart from the many clinical implications and factors, including gastrointestinal tract, mood, sleep, miscellaneous , life quality, and visuospatial and executive ability, Hcy was associated with MHs in the case of consistent baseline data. We found increased plasma Hcy levels in MH patients, especially in men, compared to NH patients. As the disease progresses and the dose of medication increases, the Hcy level of MHs also increases more significantly than that of NH. Significant correlations were observed between plasma Hcy levels and some symptoms in the PD-MH group, including motor dysfunction, cognition subdomain impairment, mood, gastrointestinal tract symptoms, cardiovascular disorder, and sleep quality. To the best of our knowledge, this work is the first to focus on PD-MH patients, explore changes in the plasma levels of Hcy, and evaluate the potential relationships between Hcy levels and certain motor and nonmotor symptoms in patients with MH. Our findings suggested that plasma Hcy may underlie the pathophysiological mechanisms of MHs and could be used to improve health-related life quality of these patients.MHs are the most frequent and earliest type of psychotic phenomena in PD, and they can appear throughout the course of PD disease, even before the onset of motor symptoms , 23. TheNutritional factors and lifestyle: excessive consumption of folate and vitamin B or too little intake of these nutrients helps elevate Hcy level. In addition, smoking, alcohol consumption, and physical inactivity are factors associated with elevated plasma Hcy level.Age and gender: some studies have suggested a correlation between HHcy and advanced age and male gender –27.Diseases and medicines: different pathological conditions, such as diabetes, hypertension, malignant tumor, and hepatic and renal insufficiency –31, as wGenetic factors: gene defects that affect the expression of various enzymes in the one-carbon cycle affect Hcy , 25.Many factors can affect the concentration of Hcy:Although many reports have discussed and confirmed the relationships between Hcy and PD or psychiatry disease separately , little Furthermore, we found that the gender difference of Hcy was highly pronounced in the MH patients. Meanwhile, the folate concentration was lower in males than in females. These results are consistent with previous findings , but theIn the MH patients, Hcy showed positive correlations with UPDRS III and NMS  and negative correlations with visuospatial/executive function. This result strongly implied that Hcy influences motor and nonmotor symptoms in MH patients. Our results are consistent with the findings associating Hcy with motor function and cognitive decline . IncreasConsistent with previous research, our study demonstrated that Hcy is reversely correlated with folate and VB12 , 38. SupStudies about the relationship between Hcy and LEDD have yielded inconsistent results. Some researchers believe that they are correlated , whereasSome limitations existed in our research. First, the number of patients was relatively small because of the strict inclusion criteria, and a referral bias might have influenced the results. Second, this work is a cross-sectional study. In a certain period of time, the continuous observation of patients' symptoms and plasma Hcy concentration is beneficial to understand the relationship and changes between the two. Third, the severity of certain nonmotor symptoms was not assessed using specific and detailed scales.MHs exert important clinical and prognostic effects on PD. Despite similar demographics, medications, and motor symptoms, PD patients with MHs showed a greater burden of nonmotor symptoms and a declined quality of life compared with PD patients without any hallucinations. Plasma Hcy levels were relatively high in MH patients. The high levels of Hcy may be a cause of other comorbid symptoms in these patients, and it explained the pathophysiological mechanisms of disease development. Hcy may be a marker of MHs and will increase the detection rate of MHs, thereby making up for the shortcomings of the scale. Whether Hcy can be a possible target in the treatment of MHs requires further study.


Aphid species that have widened their ranges due to climate change, like other insects, must adapt to low temperatures to avoid death. The metabolic changes in the body of the aphid that enabled it to survive the low temperatures of winter were investigated. Features, such as reducing their metabolic rate, an accumulation of sugars and polyols, and increased levels of some amino acids and fatty acids allow the aphids to overwinter as active-stage on the plant.Cinara tujafilina, we investigated the metabolic changes in the body of the aphid that enabled it to survive the low temperatures of winter. Using GC–MS analysis, differences in the chemical composition of the aphids in summer and winter were found. The metabolic changes were mainly related to the increased activity of the pathways of carbohydrate metabolism, such as glycolysis and the pentose phosphate pathway; a decrease in tricarboxylic acid cycle (TCA); accumulation of polyols; and increased levels of proline, tyrosine, and fatty acids.Climate changes enable thermophilic insect species to expand their ranges, but also force them to adapt to unfavourable environmental conditions in new habitats. Focusing on  Climate change allows insects to expand their ranges ,2,3 but Cinara tujafilina  , a native species of warm regions of Asia, which was introduced to the temperate regions of Europe [Aphid species that have widened their ranges due to climate change, like other insects, must adapt to low temperatures to avoid death. This is especially true of species that reproduce by parthenogenesis throughout the whole year, omitting the egg stage. One of such species is aphid f Europe . This spf Europe . Our pref Europe ,16. The C. tujafilina aphids came from cultures grown at the University of Rzeszow (Poland). Aphids were grown on Thuja orientalis plants under controlled conditions in climatic chambers . Before starting the experiment, the plants with adult wingless females were kept at 20 °C to acclimatise for 2 weeks. During experiment, aphids were kept at a temperature of 20 ± 1 °C, humidity 60 ± 5%, and with a photoperiod of L/D = 16:8 to simulate summer conditions, and at a temperature of 5 ± 1 °C, humidity 60 ± 5%, and with a photoperiod of L/D = 8:16 to simulate winter conditions for 1 month. This range of temperatures was chosen because at temperatures below 10 °C, aphids of this species migrate to the stem and root of the plant where they develop a winter form [ter form . We selem/z range: 50–850; in EI positive mode, electron energy was set to 70 eV.Sample preparation was done according to publication of Michaund and Denlinger . A totalRaw data files were converted to abf format for analysis using MSDial software (v. 3.96). To eliminate the retention time (Rt) shift and to determine the retention indexes (RI) for each compound, we implemented the correction against alkane series mixture (C-10 to C-36) directly in MS Dial. For compound identification, MSP database from CompMS site containing 28,220 records was used. Identified artifacts  were excluded from further analyses. Obtained normalized  approach) results were then exported to Excel for pre-formatting and then used for statistical analyses.t-test was performed, and metabolites with a p-value of < 0.05 were considered differential metabolites between the summer and winter groups of aphids. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) were performed to show similarities between winter and summer samples. All statistical analyses were done using Statistica version 13 programme  and PAST 4.0 software  [A  Norway) . GC–MS analysis identified 180 chemical compounds present in the body of aphids. Significant metabolic changes in the body composition of aphids in summer and winter were found in 46 chemical compounds, as shown in D-glucose 6-phosphate, glucose-1-phosphate, fructose 6-phosphate 2, D-fructose 6-phosphate, ribose)  . The sum ribose) . Winter C. tujafilina are presented in C. tujafilina in winter and summer  of metabolomic samples from summer and winter samples of d summer .p > 0.05), but changes in the amount of individual amino acids were observed  analysis showed changes in the qualitative and quantitative composition of free amino acids that occur in aphid organisms during the winter . The numSarcophaga bullata [Macrosiphum euphorbiae after heat and radiation stress [One of the main mechanisms of counteracting the effects of low temperature on an insect’s body is the activation of protective metabolic processes. The main pathways that are activated during cold stress are the carbohydrate metabolism pathways such as glycolysis, gluconeogenesis, and the pentose phosphate pathway, hence the increased level of sugars necessary for these processes, as observed in winter feeding aphids. Similar processes were also observed in  bullata ,20. In an stress . The decn stress ,22.Bravicoryne brassicae, Schizaphis graminum, Diuraphis noxia, and Aphis gossypii mainly revealed in their body the presence of mannitol, trehalose, and myo-inositol [C. tujafilina showed an increase of five compounds  and a decrease in fructose levels [The increase synthesis of sugars and polyols, which have the effect of cryoprotectants, was observed in the tissues of many species of insects. The most common metabolites with protective properties are the fructose and trehalose and polyols: glycerol, sorbitol, erythritol, and threitol ,23. The inositol ,26,27. Te levels . Our cure levels ,23, but Drosophila species subjected to heat stress [Alphitobius diaperinus, Cryptolestes ferrugineus, and Sitophilus granarius [Kermania pistacilla larvae, where the total amount of free amino acids was highest in November and then gradually decreased [Our research showed the total amount of free amino acids was slightly higher in summer samples than winter samples , but significant changes were observed in the amount of individual amino acids. Similar results were obtained in studies of beetles in which the level of free amino acids decreased as the temperature decreased ,13,28, bt stress . Both DOt stress . The inct stress  and is at stress . Accumult stress . Accumult stress . Earliert stress . An incrranarius ,12. An iecreased . The incecreased ,38. Becaecreased , the levPyrrhocoris apterus, while in Ostrinia nubilalis, palmitoleic and oleic acids were found. Increased levels of oleic acid were also found in the bodies of the aphids we studied, which suggests that it protects the cell membranes of the aphids from changing from a liquid to a solid state [Sarcophaga crassipalpis [Lipids such as unsaturated fatty acids play an important role in the processes of adaptation of insects to lower temperatures, the function of which is to maintain the stabilisation of cell membranes . Increasid state ,42. Oleisipalpis .K. pistilla indicates that this insect maintains the stability of biochemical composition, which means that it goes into a quiescent state, not diapause, and uses mainly the glycogen to fuel energy metabolism [C. tujafilina was significantly higher under cold conditions [Epiblema scudderiana moths [Lipids may also be used by insects as a source of energy during the suppressed metabolism under cold conditions. Many insects prepare for wintering through a strategy of storing energy in the form of lipids, most often in the form of triglycerides . The instabolism . Most ovtabolism . Previounditions . It was nditions . Winterina moths .Buchnera aphidicola. Facultative symbionts could affect the metabolism of the host, participating in host specialisation; rescuing the host from heat damage; and, last but not least, providing defence against natural enemies [Aphis fabae results in high aphid mortality and a low level of amino acid digestibility from the diet, which results in poor insect growth [Aphids’ symbionts play an important role in responding to environmental stress. In aphids, there are two categories of symbionts: obligate and facultative endosymbionts. The presence of obligate endosymbionts in aphids is related to their diet because aphids’ diet is deficient in many nutrients, principally amino acids, which cannot be synthesised by them and must be provided by their endosymbiont, such as  enemies ,48. It ht growth .Acyrthosiphon pisum) associated with Serratia symbiotica [N-acetyl-D-mannosamine, a precursor to sialic acid, which is a metabolite that protects against temperature stress produced by S. symbiotica [C. tujafilina under cold stress, which suggests its protective function. In amino acid biosysthesis, homoserine was also involved, the increase of which suggests synthesis of threonine and izoleucine in an overwintering aphid’s body. During cold stress, the insects also activated pathways related to oxidative stress. This was related to the emergence of reactive oxygen species (ROS), which can negatively affect insect cells. The ongoing process of oxidative stress may be indicated by an increase in the level of 2-ketoglucose dimethyl acetal, which is formed due to the addition of hydroxyl radicals [In response to the stress caused by the low temperature in the aphids’ bodies, we found an increase in shikimate acid. The presence of this acid is probably related to the symbiotic organisms that are present in the bodies of aphids and are involved in phenyloalanine, tyrosine, and tryptophan biosynthesis . The prembiotica . Facultambiotica . An elevradicals .C. tujafilina differ significantly from the summer generations, mainly due to the lower average fecundity of females, increased longevity, decreased rate of reproduction, and shortened reproductive period. However, it was shown that the biological parameters of this species do not differ significantly from the generations observed in spring and autumn on the above-ground parts of the plant [On the basis of the analysis of developmental parameters, we found that the overwintering generations of he plant ,51. ThisOur research is the first to show the metabolic response of aphids in order to adapt to the low temperatures of winter. We showed that the metabolic profile of aphid bodies differed in summer and winter. The metabolic changes were mainly related to carbohydrate metabolism pathways such as glycolysis and the pentose phosphate pathway, a decrease in activity of TCA cycle, an accumulation of polyols, increased levels of proline and tyrosine, and increased levels of unsaturated and saturated fatty acids. By reducing their metabolic rate, aphids gain an extension to their survival time, thanks to the slow catabolism of stored lipids.


Background: Irisin, a new exercise-related myokine, has been shown to be associated with a variety of diseases including serious neurological disorders. However, whether irisin is involved in the pathogenesis of vascular dementia (VD) has not yet been reported. Our aim is to determine the serum irisin level in patients with VD and investigate its relationship with cognitive function.Methods: The subjects of the study were VD patients and controls with normal cognitive function who were hospitalized in the Neck-Shoulder and Lumbocrural Pain Hospital of Shandong First Medical University from July 2018 to June 2020. Upon admission, a cognitive function assessment was performed. Enzyme-linked immunosorbent assay (ELISA) was used to determine the concentration of irisin in serum.Results: During the study period, 187 subjects (82 controls and 105 VD patients) were included in the analysis. The serum irisin level of VD patients was significantly lower than that of the control group (p < 0.001). Spearman analysis showed that irisin was positively correlated with HLD-C and MoCA, and negatively correlated with all clinical characteristics except for HCY. Logistic regression analysis showed that after adjusting for all clinical characteristics, the serum irisin of VD patients still had a significant correlation with MoCA . According to receiver operating characteristic (ROC) curve analysis, the diagnostic accuracy for serum irisin levels on VD was 76% with the sensitivity and 71% with specificity respectively.Conclusions: These data indicate that a decrease in serum irisin levels is a powerful biological marker for cognitive decline in patients with VD, even after adjustment for risk factors. Further multi-center studies need to confirm this connection, which may pave the way for new treatment options. Vascular dementia (VD) is generally considered to be the second largest subtype of dementia , 2. It aIrisin, first described by Böstrom et al. in 2012, is a small polypeptide with 123 amino acids released into the blood from the membrane protein fibronectin type III domain-containing protein 5 (FNDC5), which is cleaved at the carboxy terminus by protease in skeletal muscle . FNDC5 iInitial studies have shown that the expression of irisin is related to obesity, type 2 diabetes, and metabolic syndrome . More stThe study population were VD patients and controls with normal cognitive function who were hospitalized in the Neck-Shoulder and Lumbocrural Pain Hospital of Shandong First Medical University from July 2018 to June 2020 . VD was The Montreal Cognitive Assessment (MoCA) is a simple cognitive screening tool, which is widely used in many countries around the world with good psychometric characteristics and excellent sensitivity. MoCA is a 30-point screening tool that takes about 10 min to evaluate attention, orientation, language, memory, visual space, and executive function. The recommended normal range of MoCA is 26–30 points , 17. TheAll blood samples were collected within 24 h of admission in a fasting state and rested for at least 10 min. The blood sample was centrifuged at 1,200 g for 20 min, and the separated serum was aliquoted and stored at −80°C for later use , 19. Stat-test. Spearman rank correlation was used for bivariate correlation. The relationship between serum irisin concentration and clinical characteristics was evaluated by logistic stepwise regression analysis. Sensitivity and specificity of measured variable for VD diagnosis were determined by receiver operating characteristic (ROC). All statistical analyses were performed using SPSS for Windows, version 23.0 . Statistical significance is defined as two-tailed P < 0.05.The results of categorical variables are represented by n or percentages, and the results of continuous variables are represented by mean ± standard deviation (SD). Categorical variables are compared by chi-square test, while continuous variables are analyzed by student's p > 0.05). The serum irisin concentrations of the control group and the VD group were (127.80 ± 11.15) and (79.37 ± 8.74) ng/ml, respectively, and there are significant differences between the two groups (p < 0.001).A total of 82 normal cognitive function controls and 105 VD patients were enrolled in this study. The baseline clinical characteristics of all subjects are shown in r = −0.231, p < 0.001), gender , BMI , SBP , DBP , TC , TG , LDL-C  and FBG (r = −0.170. p < 0.001), and positively correlated with HDL-C  and MoCA scores . There was no significant correlation between serum irisin and HCY in patients with VD . The results of the Spearman rank correlation test are shown in In the Spearman rank correlation test, the serum irisin of VD patients was significantly negatively correlated with age (p < 0.001). After further controlling of SBP, DBP, TC, TG, LDL-C, HDL-C and FBG in Model 2, the serum irisin concentration of VD patients still has a significant correlation with the MoCA score . HCY is considered to be an important factor affecting cognitive function, we pulled it into Model 3. Finally, after controlling of all baseline clinical characteristics in Model 3, the correlation between serum irisin concentration and MoCA score in VD patients is still significant .The logistic regression analysis results are shown in To evaluate the diagnostic value of serum irisin levels as a potential marker for VD, ROC curve analysis was performed . The areThis is the first time to study the relationship between cognitive decline and serum irisin concentration in VD patients. Our results show that the serum irisin concentration of VD patients is significantly lower than that of controls with normal cognitive function, and there is a significant positive correlation with the MoCA score. This relationship still exists after adjusting the clinical baseline data, suggesting that serum irisin may be used as a biomarker of cognitive function for VD.In recent years, irisin has been confirmed to be related to the onset of many diseases. Yeon-Kyung Choi et al. found that serum irisin concentration was lower in newly-onset type 2 diabetes patients and is affected by blood sugar levels, suggesting that irisin played an important role in the pathogenesis of type 2 diabetes . The stuMore and more evidences showed that irisin could also participate in the pathogenesis of a series of neurological diseases. Wen-Jun Tu's team found that serum irisin level was a powerful biological marker for the risk of post-stroke depression . The teaOur research has some limitations. First of all, our research subjects come from a single region and the sample size is relatively small. Secondly, exercise can affect the level of irisin . We did in vivo and in vitro are needed to further confirm this connection, which may provide a novel treatment for VD.This study shows that a decrease in serum irisin level is a potential biological marker of cognitive impairment in VD patients, even after adjustment for confounding factors. Future studies The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the Ethics Committee of the Neck-Shoulder and Lumbocrural Pain Hospital of Shandong First Medical University. The patients/participants provided written informed consent to participate in this study.YZ designed this research and wrote the manuscript. BS, CW, FZ, GuangjH, and GuangsH participated in data collection, experimental process, and data analysis. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.