Polymer composites are favorite materials for sensing applications due to their low cost and easy fabrication. In the current study, composite nanofibers consisting of polyethylene oxide (PEO), oxidized multi-walled carbon nanotubes (MWCNT) and copper oxide (CuO) nanoparticles with 1% and 3% of fillers  were successfully developed through electrospinning for humidity sensing applications. The composite nanofibers were characterized by FTIR, XRD, SEM and EDX analysis. Firstly, they were loaded on an interdigitated electrode (IDE), and then the humidity sensing efficiency was investigated through a digital LCR meter (E4980) at different frequencies (100 Hz–1 MHz), as well as the percentage of relative humidity (RH). The results indicated that the composite nanofibers containing 1% and 3% MWCNT, combined with CuO in PEO polymer matrix, showed potent resistive and capacitive response along with high sensitivity to humidity at room temperature in an RH range of 30–90%. More specifically, the PEO–CuO–MWCNT: 1% nanocomposite displayed a resistive rapid response time within 3 s and a long recovery time of 22 s, while the PEO–CuO–MWCNT: 3% one exhibited 20 s and 11 s between the same RH range, respectively. Recently, tremendous efforts have been made to improve the performance of chemical and physical sensors for technological and daily life applications . In some2, ZnO, SnO2, Al2O3, CuO) [Moreover, polymer-based humidity sensors offer high sensitivity, small hysteresis, power efficiency, flexibility, low cost and versatile applications. The performance of humidity sensors relies on the sensitivity of sensing materials, which is closely associated with their chemical structure and specific functions. Hence, the sensitivity enhancement of these sensing materials towards humidity is challenging . Very reO3, CuO) ,8, carboO3, CuO)  and hydrO3, CuO) , polyethO3, CuO) , polyvinO3, CuO)  and polyO3, CuO) ) have beOver the years, the incorporation of inorganic nanomaterials into polymers has been extensively reported. Metals and metal oxide nanostructures have been paid much attention in the development of sensing devices due to their divergent physical and chemical properties . Metal oOn other hand, CNTs have been applied for humidity sensing more widely compared to other carbon-based materials due to their superior electrical, physical and chemical properties. In addition, CNTs possess a large surface-to-volume ratio and nanoscale structure with a hollow core, and hence they are capable of adsorbing large amounts of foreign molecules on the surface . CapacitA significant variation of permittivity and conductivity among polymer-based humidity sensors can be improved by controlling the porosity and the sensing layer of the polymer matrix . Thanks Recently, miniaturized sensors for portable devices such as mobile phones are available, including the following: SHTC1 humidity sensor from Sensirion and LPS331AP pressure sensor from STMicroelectronics are being used in Samsung cell phones; Bosch Sensortec company introduced a miniaturized humidity sensor capable of sensing pressure and temperature BME280), which are based on Micro-Electro Mechanical Systems (MEMS) . Althoug0, which The present work demonstrates the realization of low-cost, efficient, flexible and highly stable humidity sensors based on polyethylene oxide, CuO and MWCNTs composite nanofibers by electrospinning. The morphology and structure of composite nanofibers were investigated by SEM, FTIR, EDX and XRD analysis. The humidity sensing performance of the prepared PEO-CuO-MWCNT composite nanofibers was studied through an LCR meter  at fluctuating frequency and RH conditions. The said sensors showed a high capacitive response to humidity, exhibiting high sensitivity, good linearity and fast recovery and response times. Thanks to these remarkable properties, the sensors can be advantageous in several applications such as the monitoring of health and medical facilities, environmental measurements, engineering instruments and remote control of various electronic devices.2SO4) 95% and nitric acid (HNO3) 70% were purchased from Merck KGaA; Darmstadt, Germany.PEO powder  with viscosity average molecular weight of 30,000 g/mol. and MWCNTs  were provided by National Centre for Physics, Quaid-e-Azam University, Islamabad, Pakistan. Sulfuric acid  was dissolved in a calculated volume of distilled water to prepare a solution (0.2 M). The CuSO4 solution was taken in a beaker and placed on a magnetic stirring hot plate, with a pH electrode inserted in the solution. Then, a solution of NaOH (2.0 M) was added dropwise into the CuSO4 solution under continuous stirring. The addition of NaOH was continuously done until a pH of 8.5 was obtained. The solution was further stirred at 90 °C for 1 h, and then the resultant precipitate was recovered by filtration followed by washing with distilled water. The precipitate was calcined in a muffle furnace at 400 °C for 4 h. Finally, a dark brown powder of CuO was obtained, which was cooled in a desiccator and stored in vials.CuO nanomaterials were successfully prepared according to a precipitation method . A stoic2SO4 and HNO3 (3:1) and sonicated in an ultrasonic bath for 3 h at 40 °C. The suspension was recovered by means of filtration and washed with distilled water until the pH to be neutral. The MWCNTs were then dried under vacuum in an oven at 50 °C for 5 h. MWCNTs were oxidized by an acid treatment using a procedure reported elsewhere . MWCNTs The PEO-CuO-MWCNT composite blend was prepared by incorporating CuO nanoparticles and oxidized MWCNTs as fillers in PEO polymer matrix. To prepare blend solution, PEO was initially dissolved in distilled water and stirred for 1 h, and then the calculated weights of CuO nanoparticles and MWCNTs required for a ratio of 1:1 and 1:3  were added to the PEO solution, respectively. The mixture was homogenized by ultrasonication for approximately 4 h at 40 °C. Finally, the resulting homogenous dispersion with the dark brown appearance was continuously stirred to get a viscoelastic solution for the synthesis of nanofibers .The composite nanofibers were synthesized by electrospinning. The PEO–CuO–MWCNT solution was loaded in a 10 mL syringe fixed with a 21 gauge blunt needle linked to a high-voltage power supply. The positive electrode of the power supply was connected to the needle, while the negative one was connected to an interdigitated electrode (IDE). Aluminum foil was also attached to the negative electrode in order to collect the nanofiber for further characterization. The distance between the positive electrode (syringe) and the collector was 15 cm, and high electrostatic voltage was applied to synthesize the nanofibers at a flow rate of 0.5 mL/h by a syringe pump leading to the formation of a droplet of the composite solution. At a voltage of 15 Kv, the solution was sprayed on the Al foil and IDE screen, composite nanofibers were formed as the solvent evaporated. Graphical presentation is given in XRD patterns of pure PEO, CuO nanoparticles, MWCNTs and PEO–CuO–MWCNT nanofibers were performed using an X-ray diffractometer  at room temperature. The morphology of the samples was examined by scanning electron microscope , while the elemental analysis of the composite and the precursor materials was investigated by EDS analysis . Fourier transform infrared (FTIR) spectra of pure PEO, CuO nanoparticles, MWCNTs and nanocomposites were taken through an FTIR spectrophotometer . The humidity sensing properties of the composite nanofibers were studied by measuring their capacitance and resistance at different RH levels using a digital LCR meter . The reference humidity sensor (DHT11) was combined with a microcontroller , and both were connected to a computer through an RS232 card. The composite nanofibers were loaded on the interdigitated electrode (IDE) and connected to the LCR meter. The IDE electrode was placed in a moisture chamber along with the reference sensor, while atmospheres containing various humidity levels were adjusted passing dry nitrogen and air saturated with vapors of deionized water. The moisture chamber was sealed, and the change in resistance and capacitance was recorded at every 60 s under varying RH between 30 and 90%. The values of the resistance and capacitance changes with % RH were recorded and digitized, respectively.The morphological features of the composite nanofibers were examined by Scanning Electron Microscopy (SEM). The X-ray diffraction (XRD) patterns of pure PEO, CuO nanoparticles, MWCNTs and composite nanofibers are illustrated in In the XRD patterns of PEO–CuO–MWCNT: 1% and PEO–CuO–MWCNT: 3% , the cha−1, which is related to asymmetric C-H stretching vibrations of CH2 groups. The significant central peak at 1468.33 cm−1 is due to asymmetric bending vibrations of CH2 groups, while a strong intense peak at 1102.06 cm−1 is attributed to the asymmetric C–O–C stretching for epoxide groups. These configurations are in agreement with the structural features of PEO polymer [−1 [−1, representing the O–H and C=O of carboxylic acids, respectively [The FTIR spectra of the polymer matrix, i.e., PEO, fillers (CuO and MWCNTs) and the composite nanofibers, are shown in  polymer . The FTIymer [−1 . The speectively , this in−1 is slightly decreased, which may be attributed to the interaction between the ether group of the poly ethylene oxide and the hydroxyl one of the oxidized MWCNT, which are linked through hydrogen bonding [The spectra of PEO–CuO–MWCNT: 1% and PEO–CuO–MWCNT: 3% nanocomposites are displayed in  bonding . Since t bonding .The humidity sensing performance of PEO–CuO–MWCNT composite nanofibers was investigated by measuring its capacitance and resistivity response in various levels of RH as a function of frequency. Initial experiments showed that PEO– composite nanofibers with 1% CuO presented a significant resistivity response, better than the respective capacitance response upon changing the RH. 0 or C0.It has been shown that frequency has a major role and impact on the humidity sensing performance. The results of humidity sensing efficiency of the composite nanofibers are investigated at various input frequencies such as 1 KHz, 10 kHz, 100 KHz and 1 MHz. 2O molecules to MWCNTs. The higher the amount of RH, the greater the adsorption of H2O molecules and the more electrons are transferred. The PEO–MWCNT–CuO behaves as a semiconductor, containing electrons (−vie charges) and holes (+ive charges). In the present case, the electrons are minority carriers and the holes are the majority carrier. As electron (minority carrier) from water molecules are transferred, this will lower the concentration of majority carrier, i.e., +ive charges (holes), thereby resulting in an increase in the resistance of the sensor [Our results showed that the resistance and capacitance of the sensor significantly increased when RH value changed from 30–90% RH. This significant increase in capacitance could be associated with the adsorption of water molecules on the surface of the sensor, which directly depends on the RH level . In the e sensor . 5 Ω, which is smaller than 6.7 × 106 Ω at 90% RH at 1 KHz frequency [2) as compared to PEO–MWCNT–CuO: 3% nanocomposite-based sensor, as shown in The remarkable increase of resistivity response was measured over a % RH range from 30 to 90% at four different frequencies, i.e., 1 KHz, 10 KHz, 100 KHz and 1 MHz, respectively. It was observed that the resistance at lower frequencies (1 KHz and 10 KHz) enhances proportionally to RH. This resistance change of the sensor is displayed in requency . At low −10 F (for 30% RH) to 7.9 × 10−8 F (for 90% RH) as shown in 0 and 0 presents the permittivity in free space.The capacitance response of PEO–CuO–MWCNT: 3% nanocomposites drastically increased from 1.09 × 102) with a positive slop over wide humidity range levels such as 30–90% RH as displayed in From Equation (3), it can be concluded that the capacitance of the sensing material is directly proportional to γ and inversely proportional to frequency . MoreoveOne of the considerable parameters for the fabrication of excellent and economical humidity sensors is their fast response and recovery time. Usually, response and recovery times are measured in 10–90% RH, but in the present case, the response of the sensor below 30% RH was negligible; hence, measurement was carried out between 30 and 90% RH. When air humidity is suddenly varied from 30 to 90% RH, the response and recovery time can be verified by the capacitive and resistive variation of the humidity sensor. The response or recovery time is the time responsible for causing a 90% change of their original value when increasing the humidity from a low to a high degree or vice versa, respectively. Due to the hydrophilic nature of the PEO matric, and its large surface area, it is perceived that composite nanofibers allow faster adsorption of water molecules. In this context, the response and recovery time of resistivity and capacitance response of PEO−CuO−MWCNT: 1% and PEO−CuO−MWCNT: 3% composite nanofibers have been presented in Owing to their low density and high aspect ratio, carbon nanotubes have proven to be ideal fillers for designing diverse polymer composites with enhanced mechanical performance, high electrical conductance and multi-functional properties . AdditioThe humidity sensing mechanism for the adsorption of water molecules on the surface of PEO−CuO−MWCNTs is displayed in To explain the humidity sensing mechanism, two types of adsorption interactions were considered i.e., chemisorption and physisorption . The curA novel highly responsive humidity sensor was fabricated based on PEO−CuO−MWCNTs composite nanofibers. The composites nanofibers were developed for capacitive and resistive humidity applications prepared via electrospinning. The composite nanofibers PEO−CuO−MWCNT: 1% and PEO−CuO−MWCNT: 3% humidity sensor revealed high sensitivity with fast response and recovery time. We found that the PEO−CuO−MWCNT: 1% composite nanofibers showed high humidity sensitivity (3798.2%) and exhibited a quick response of 3 s and recovery time of 22 s. It was observed that in PEO−CuO−MWCNT: 1% composite nanofibers, the resistance increased linearly when exposed to humidity. The PEO−CuO−MWCNT: 3% nanocomposite showed a response of 20s and a rapid recovery time of 11s with high sensitivity , while the capacitance increased as the frequency decreased. All results demonstrate that increasing the concentration of fillers i.e., MWCNTs and CuO nanoparticles is an effective approach to improve the sensing properties. The proposed method allows the scalable production of multicomponent sensor devices based on polymer matrix composite with high stability, flexibility and versatility for several industrial applications.


ABCA1 promoter methylation level may result in the progression of coronary artery disease. Thus, the present study investigated the association between promoter methylation status of ABCA1 and inflammation in the development of premature coronary artery disease (pCAD).ATP-binding cassette transporter A1 (ABCA1) plays a major role in high-density lipoprotein (HDL) metabolism and reverse cholesterol transport (RCT) and exerts anti-inflammatory effects. Increased n = 90 each) were recruited from the Characteristic Medical Center of the Chinese People's Armed Police Force from June to December 2019. Using pyrosequencing, the levels of ABCA1 promoter methylation in their blood samples were evaluated. Serum concentrations of lipids, interleukin 1β (IL-1β), C-reactive protein (CRP), and circulating free DNA/Neutrophil extracellular traps (cfDNA/NETs) were also routinely measured and compared between the two groups. P values < 0.05 were considered statistically significant.PCAD patients and healthy individuals . Based on binary logistic regression analysis, ABCA1 promoter methylation level was identified as an independent risk factor for pCAD development . Furthermore, ABCA1 promoter methylation levels were negatively correlated with HDL levels  and positively correlated with the levels of CRP, cfDNA/NETs, and IL-1β . Multiple regression analysis showed that the serum levels of CRP, IL-1β, and cfDNA/NETs independently affect ABCA1 promoter methylation.The mean ABCA1 promoter are associated with low HDL cholesterol levels and an increased risk of pCAD. Inflammatory factors and NETs may be involved in the progression of pCAD by affecting ABCA1 promoter methylation levels.Our findings indicate that high methylation levels at the  Despite the continuous improvement in preventive measures and medical technology, coronary artery disease (CAD) remains as the main cause of mortality worldwide , 2. Its ABCA1 gene is located on chr9q31, having a total length of 149 kb and including 50 exons and 49 introns [Various genetic and environmental factors have been identified to contribute to pCAD development ; in part introns . It medi introns .ABCA1 promoter have been inconsistent and conflicting [DNA methylation is an epigenetic phenomenon that typically inhibits gene expression and is characterised by the introduction of an activated methyl group onto the fifth carbon atom of cytosine catalysed by DNA methyltransferase (DNMT) . In receflicting , 15.Inflammatory response is closely related to the occurrence and development of AS . ABCA1 hABCA1 promoter.Neutrophil extracellular traps (NETs) are extracellular fibres composed of DNA, histones, and microbicidal granular proteins and play important roles in antimicrobial responses as they can effectively trap pathogens, thus preventing infections . AlthougABCA1 promoter influence the occurrence of pCAD and investigateing the relationship between ABCA1 promoter methylation levels with lipid levels, inflammatory factors, and NETs in the Chinese pCAD patients, which will contribute us to explore the molecular pathogenesis of pCAD.This study was aimed at examining whether methylation level of the Consecutive patients with pCAD were recruited from the Department of Cardiology in the Characteristic Medical Center of the Chinese People's Armed Police Force from June to December 2019. PCAD was defined as a history of myocardial infarction, angioplasty, revascularisation surgery, or more than 50% stenosis in at least one coronary artery determined by coronary artery angiography and diagnosed before age 55 in men and 65 in women. The exclusion criteria included patients with cardiogenic shock, severe heart failure, severe liver and kidney dysfunction, a history of malignant tumour, and those undergoing treatment with lipid-lowering drugs or other drugs that affect blood lipid levels in the past 3 months. Finally, 90 pCAD patients were enrolled , and 25 patients were screened. Meanwhile, 112 sex- and age-matched asymptomatic individuals with no family history of CAD were recruited from the health examination department of our centre. Upon examination by coronary artery computed tomography or coronary angiography, we excluded 22 participants who at risk of progression to pCAD due to the coronary lesions < 50%. Finally, 90 individuals with normal coronary arteries were included in the control group.The study was conducted in accordance with the Declaration of Helsinki and approved by the Ethics Committee of the Characteristic Medical Center of Chinese People's Armed Police Force .All participants underwent general health examination to measure height, weight, waist circumference, body mass index (BMI), and detailed information on medical (diabetes and hypertension) and treatment history and smoking habits were collected. Participants regularly smoking tobacco products at least once a day or those that have smoked 30 days prior to admission were considered as current smokers. Hypertension was diagnosed as systolic blood pressure ≥ 140 mmHg and/or diastolic ≥ 90 mmHg and/or undergoing antihypertensive treatment. Diabetes mellitus was considered when participants are taking glucose-lowering drugs due a previous diagnosis of diabetes according to the American Diabetes Association criteria . FurtherVenous blood samples were collected after an overnight fast of at least 8 h and assessed at the laboratory of our centre. Plasma glucose, TC, TG, HDL-C, and LDL-C were enzymatically assayed using a Siemens automatic biochemical analyser .ABCA1 [Pyrosequencing was conducted as previously described to detect the methylation status of the CpG islands in the promoter region of ABCA1 , 24. PriThe ABCA1 concentration in the blood samples was determined using an enzyme-linked immunosorbent assay (ELISA) kit  following the manufacturer's instructions. The average optical density of each sample was determined using an xMark™ Microplate Spectrophotometer ; the four-parameter equation was fitted; and the serum ABCA1 concentration in each sample was calculated based on the equation of the standard curve.CRP was determined by turbidimetric immunoassay performed on an automated-analyser  using a CRP diagnostic kit . The level of IL-1β was detected by ELISA assay using a commercially available kit .NETs comprise circulating free DNA (cfDNA), which can activate and amplify the immune response to an immunogen in a proinflammatory manner. Therefore, cfDNA can be used as a surrogate marker to assess the extent of inflammation in the body. The cfDNA/NETs component was measured as previously described . Quant-it-test, whereas non-normally distributed data are expressed as the median and interquartile spacing and analysed using the Mann–Whitney test. Categorical variables were compared using Chi-square test. Correlation was evaluated using the Pearson's and nonparametric Spearman's rank correlation test. The independent association of each risk factor with pCAD was analysed using binary logistic regression. Factors affecting the methylation of ABCA1 promoter were analysed using multiple regression analysis. P values < 0.05 were considered statistically significant.SPSS19.0 software was utilised for statistical analysis. Normality was assessed using the Kolmogorov–Smirnov test. Normally distributed data are expressed as the mean ± SD and analysed using an independent P > 0.05). The levels of HDL-C were significantly lower  in the pCAD group than those in the control group, whereas BMI, waist circumference and HbA1c levels were significantly higher in the pCAD group (Table vs. pCAD: 5.1 (2.3–7.4) years) and the rates of insulin (control: 22.2% vs. pCAD: 24.4%) and hypoglycaemic drug (control: 26.7% vs. pCAD: 33.3%) used between the two groups (P > 0.05).There were no significant differences in age, sex, smoking rate, systolic and diastolic blood pressure, TC, TG, LDL-C, fasting blood glucose, homocysteine (Hcy), and the incidence of hypertension between the two groups , whereas the serum ABCA1 concentration was lower in the pCAD group than that in the control group .Similar to the results of Guay et al. , 24, onlup Table ; and thevs. control) as the dependent variable and age, sex, BMI, waist circumference, TG, TC, HDL-C, LDL-C, HbA1c, diabetes, hypertension, and ABCA1 promoter methylation status as the covariates, we identified the methylation level of the ABCA1 promoter (odds ratio (OR) = 2.878, 95% confidence interval (CI) 1.802–4.594; P < 0.001), HDL-C , BMI  and HbA1c  as independent risk factors for pCAD development  , TC , or TG .01) Fig. a; but weABCA1-A   Fig. b–d. The ABCA1 promoter. As evident in Fig. P > 0.05).We further assessed whether age and sex are associated with the mean methylation level of the ABCA1 promoter methylation levels and plasma HDL-C levels are negatively correlated and that FH patients with CAD exhibit higher ABCA1 promoter methylation levels than those without [ABCA1 promoter methylation was significantly correlated with the age and the incidence of CAD [ABCA1 promoter methylation levels and the severity of CHD and that the effect of ABCA1 promoter methylation levels on CHD is independent from its effect on plasma lipid levels.As the main pathophysiological basis of CAD, AS manifests as chronic inflammation and the development of AS involves abnormal DNA methylation . A previ without . Moreovee of CAD . Howevere of CAD  in an InABCA1 promoter in pCAD patients were significantly higher, whereas the level of HDL-C and serum ABCA1 concentration were lower; (2) The methylation level of the ABCA1 promoter was an independent risk factor for pCAD, along with traditional risk factors, such as low HDL-C, high BMI and HbA1c; and (3) ABCA1 promoter methylation levels were negatively associated with the plasma HDL-C levels. These results imply that, on the one hand, ABCA1 promoter methylation regulates its mRNA expression by inhibiting the transcription of the ABCA1 promoter, resulting in decreased levels of ABCA1, which leads to the inactivation of RCT and the acceleration of lipid deposition. Moreover, many risk factors affect pCAD, such as smoking, dyslipidaemia, diabetes, hypertension, and family history of CAD [ABCA1 promoter increases the risk of pCAD independently of the lipid level, and the genetic predisposition may play a key role. Therefore, the methylation level of the ABCA1-A promoter may be used as a potential biomarker to predict the occurrence risk of pCAD in clinical practice and to assist in the early diagnosis, treatment, and monitoring of pCAD in a non-invasive manner.This study established the following: (1) Compared with the control group, the average methylation level of the y of CAD . We alsoDNA methylation levels are influenced by heredity, age, smoking, nutritional status, and environment –32 and cABCA1 promoter methylation level was positively associated with serum inflammatory factors  and cfDNA/NETs; meanwhile, the patients of pCAD group had higher serum levels of inflammatory cytokines and cfDNA/NETs than the control group, suggesting that inflammatory responses contribute to the promotion of pCAD. The reason may be due to the changes in the bodily environment of patients with pCAD that can cause epigenetic modifications, resulting in increased methylation of the associated genes and increased activity of DNA methyltransferases [ABCA1 promoter in pCAD patients might affect the expression of ABCA1, which interacts with inflammatory factors to stimulate the development of early coronary AS.Our study found that sferases , thus diABCA1 promoter methylation, which may verify clinically that inflammatory mediators can further promote the development of pCAD via regulating DNA methylation. Therefore, we speculate that intervention with pharmaceuticals exerting anti-inflammatory activities during the early course of pCAD may regulate the methylation status of ABCA1, and can be applied to the development of therapeutic strategies for pCAD. However, more studies are needed to confirm our hypothesis.In addition, increasing evidence shows that epigenetic processes are not static and that DNA methylation is a reversible process that can be regulated. Lou et al.  has reveABCA1 promoter methylation level. We attribute this to our study population that was limited to patients with pCAD and the relatively narrow age span. Another study has showed that the methylation levels was more significantly different in elderly patients with coronary heart disease > 65 years old than those younger [Furthermore, unlike a previous study , our stu younger , suggestABCA1 promoter due to the low ABCA1 concentration in the peripheral blood. The expression of ABCA1 mRNA need to be further investigated. Third, as the definition of pCAD is not uniformly established, and consider the actual situation of our centre and various literature studies [First, due to the specificity of the age of onset of pCAD and the high cost of pyrosequencing analysis, our sample size was relatively small, and our participants were recruited from a single centre within a stable population. Second, we did not find any correlation between the serum ABCA1 concentration and the methylation level of  studies –7, we usThe incidence of pCAD has increased significantly along with improved living standards; however, the pathogenesis of pCAD is not well understood. In the future, a larger sample size is required for the assessment of the potential predictors of severity of pCAD and comparison with mature CAD. Meanwhile, further basic and clinical studies are warranted to confirm the role of inflammation on methylation.ABCA1 promoter and lower HDL-C plasma level and identified high ABCA1 promoter methylation as an independent risk factor for pCAD development. Additionally, our findings suggest that inflammatory factors and NETs are involved in the progression of pCAD by affecting ABCA1 promoter methylation levels. Therefore, the regulation of DNA methylation may serve as a novel therapeutic strategy for pCAD.This study revealed the association between higher methylation levels at the


Transport interventions help to facilitate the sustainable travel behavior. The effects of transport interventions on travel choices have been addressed extensively. However, little research has been devoted to the influence of transport interventions and travel choice on travel perception. This study aimed to investigate the relationship among the three aspects. Two intervention measures, information intervention and public transport service improvement, were selected. Intervention experiments were designed to collect mode choice and corresponding travel perception in different experiment stages. Process models of information intervention and public transport service improvement were proposed. The results show that information intervention only had a minor effect on mode choice and had no direct effect on travel perception. Public transport service improvement in in-vehicle time and comfort enhanced public transport use dramatically. Comfort improvement also had positive effects on travel perception. Walking had positive and public transport trips had negative effects on travel perception. For travelers who had a high evaluation of car trips, the probability of green mode use would decrease. Travelers who gave high marks to trips by green mode would have a higher probability to keep traveling by green mode. This study contributes to facilitating public transport use and enhancing positive perception during traveling. Transport interventions can help to change travel behavior and shape travelers’ sustainable behavior. Transport interventions include soft measures, such as the dissemination of green mode traveling, and hard measures, such as license plate restriction. The implementation of transport interventions facilitates travel choice change, including mode choice, departure time choice, activity choice, etc. Travel choice has impacts on travel perception. Transport interventions brings direct or indirect effects on travel perception through influencing travel choice, and, the other way round, feedback exists among transport interventions, travel choice, and perception. If the traveling experience become worse after the travel choice change, travelers will continue adjusting their choices to obtain higher levels of travel perception. Transport managers also need to adjust intervention strategies according to travelers’ perception and travel choice behavior change. The relationship among transport interventions, travel choice behavior, and travel perception is shown in Transport interventions could facilitate sustainable travel behavior to some extent. The evaluation of transportation interventions should also focus on travelers’ perception change besides the change of travel choice behavior. At present, numerous studies have been performed to explore the influence of transport interventions on travel choice behavior. Rose and Ampt 2001) [ [1] evalThe objective of this study was to investigate the relationship between transport interventions, mode choice, and travel perception, which was used to provide recommended policy implications for sustainable travel behavior development. Soft measures including information intervention and public transport service improvement were selected as two intervention strategies in this study. The remainder of this study was organized as follows: Information intervention in the transportation field was mainly implemented to promote and encourage pro-environmental travel behavior . KnowledIn this study, information intervention experiment was designed to disseminate information about “health and environment benefits brought by non-motor vehicle traveling”, “air pollution caused by motor vehicle traveling”, and “improvements of green mode service”. Based on the information intervention experiment, the relationship model among information intervention, mode choice, and travel perception would be proposed.Public transport plays an important role in contributing to urban sustainability. Gustavo Petro, the mayor of Bogotá, ever said “A developed country is not a place where the poor have cars. It’s where the rich use public transportation”. Attractive public transport service was the contributing factor of a sustainable transport system . In receIn this study, considering the whole trip, accessibility, next bus service, bus lane, optimization of public transport network and operation would be included to reduce access–egress time, waiting time, in-vehicle time, the number of transfers, and increase comfort in vehicle.Travel perception was used to describe travelers’ overall cognitive evaluation and emotional feeling during traveling. Its synonyms included travel satisfaction , travel Compared with 20 years ago, with the increasing popularity of the automobile, the acceleration of urbanization, and improvement of material and spiritual pursuits, people’s daily activities became more diverse. Traveling for different purposes, such as work, school, shopping, socializing, and entertainment, has become an integral part of people’s daily life. With more understanding of activity-based travel, scholars thought that travel itself should not only be treated as ways to conducting activities, but as a significant activity ,34. BesiAffective Balance Scale  according to the average score of cognitive and affective evaluation. Therefore, travel perception in stage T1 was subdivided into four variables, PT_P, bike_P, walk_P, and car_P, which represented average score of travel perception by PT, bike, walk, and car was greater than 0.5 or not. Information intervention was a binary variable.Similar to Model 1, Model 3 was also a multiple linear regression model. It was used to analyze the influence of information intervention and mode choice on travel perception in stage T2 and T2′. Average score of cognitive evaluation (CE2) and average score of affective evaluation (AE2) were dependent variables. The independent variables were mode choice, whether intervention information was received in stage T2 and T2′, and individual characteristics. According to According to the results in Model 2, travel perception in stage T1 had significant effects on mode choice in stage T2 and T2′. The more positive travel perception when using public transport in stage T1, the higher the probability of using public transport and walking in stage T2 and T2′, which were 2.8 and 4.3 times, respectively, than traveling by car. The more positive travel perception for bike trips in stage T1, the higher the probability of riding and walking in stage T2 and T2′. Similarly, if participants had more positive cognitive and affective evaluation during walking, they were more likely to travel by walking in stage T2 and T2′. If travelers felt more positive during car trips, they were less likely to use public transport and bike in stage T2 and T2′. Information intervention had positive impacts on public transport use and walking. In other words, the probabilities of travelers using public transport and walking increased after information intervention. However, the influence coefficients were quite small, which, indicating that information intervention had a very limited impact on public transport use and walking. It was found that public transport, bike, and walking were closely linked. Travelers who had positive evaluation of public transport and bike trips also tended to travel by walk, but travelers who had a positive evaluation of bike and walking trips did not increase the probability of travelling by public transport. Based on the above influence relationship, it was even more important to promote public transport use and improve the service level of public transport to enhance travelers’ positive perception of public transport trips. Income level had negative effects on bike and walking. The results in Model 2 could help to predicate travelers’ mode choice after implementing information intervention and evaluate information intervention according to transport mode shift from automobile to green modes.In Model 3, compared with car trips, the influence of walking on travel perception was still positive. Public transport use had negative impacts on travel perception. Information intervention had direct influences on mode choice but no significant effects on travel perception. Age and education still had significant impacts on travel perceptionThe process model results of information intervention demonstrated that information intervention increased the probability of traveling by public transport and walking, but the impact was weak. Walking could increase positive aspects in travel perception. However, public transport trips had negative effects on travel perception. For travelers who had a high evaluation of travel perception during car trips, the probability of green mode  use would decrease. Travelers who gave high marks to travel perception during public transport traveling, bike trips and walking would have a higher probability to keep the transport mode use or traveling by other green modes. The relationship among information intervention, mode choice, and travel perception is illustrated in The data of participants’ mode choice and travel perception in recent trip (R1) and in different scenarios of public transport service improvement (R2) were collected. Individual characteristics were collected in stage T0. On the basis of these data, the process model among public transport service improvement, mode choice, and travel perception was proposed to explore their mutual relations.Mode choice in stage R1 affected participants’ travel perception. Travel perception in stage R1 had impacts on subsequent mode choice, namely mode choice in stage R2. In addition, mode choice in stage R2 was also affected by service level of public transport. Travel perception in R2 was influenced by mode choice and service level of public transport. The detailed variables and descriptions of the process model under public transport service improvement were shown in The process model of public transport service improvement in stages R1 and R2 included three specific models.Model 1 was a multiple linear regression model which described the influence of mode choice on travel perception in stage R1. There were two dependent variables in Model 1, the average score of cognitive evaluation (CE1) and the average score of affective evaluation (AE1). Both were continuous variables. The independent variable was mode choice in stage R1 and individual characteristics. Mode choice in stage R1 included four specific variables, mod1_PT (public transport), mod1_bike (bike), mod1_walk , and mod1_car (car). Car was selected as the reference mode.Model 2 was a binary logit model, which represented the impact of travel perception in stage R1 and service level of public transport on mode choice in stage R2. Dependent variable was mode choice in stage R2 . Independent variables included travel perception in stage R1, service level of public transport in stage R2, and individual characteristics. Similar to Model 3 was also a multiple linear regression model used to analyze the impacts of public transport service level and mode choice on travel perception in stage R2. Average score of cognitive evaluation (CE2) and average score of affective evaluation (AE2) were dependent variables. Independent variables were the service level of public transport and mode choice in stage R2, and individual characteristics. According to According to the results in Model 2, travel perception in stage R1 had significant effects on mode choice in stage R2. The more positive evaluation of travel perception when using public transport and bike in stage R1, the higher the probability of using public transport in subsequent stage R2. However, if travelers felt more positive during car trips in stage R1, they were less likely to use public transport in stage R2. Compared with scenario S4, scenario S1, S2, S3, S5, S6 all had significant positive effects on public transport use. The improvement aspects of public transport service level of S1, S2, S3, S5 and S6 were reflected by in-vehicle time reduction or the degree of comfort increase compared with S4, as shown in In Model 3, the regression coefficients suggested that scenario S2, S3, and S6 had positive impacts on travel perception. In scenario S2, S3, S6, there were no crowding in vehicle and every passenger had a seat. It could be concluded that increased comfort in vehicle improve passengers’ travel perception significantly. Compared with traveling by car, public transport trips had negative effects on the affective evaluation of travel perception. It meant that car travelers felt more pleasant and enjoy traveling compared with traveling by public transport. Age and education still had significant impacts on travel perceptionThe process model results of public transport service improvement demonstrated that the improvement of public transport service significantly increased the probability of public transport use. Compared with car trips, walking could increase positive aspects in travel perception; however, public transport trips had negative effects on travel perception. For travelers who had a high evaluation of travel perception during car trips, the probability of public transport use would decrease. Travelers who gave high marks to travel perception during public transport traveling and bicycle trips would have a higher probability to travel by public transport. The relationship between public transport service improvement, mode choice, and travel perception is illustrated in This study focused on the influence of different transport intervention strategies, including information intervention and public transport service improvement on mode choice and travel perception. Based on the collected mode choice and travel perception data under these two intervention strategies, process models of information intervention and public transport service improvement were proposed to explore the relationship among transport interventions, mode choice, and travel perception.The results from the process model of information intervention demonstrate that information intervention increased the probability of traveling by public transport and walking, but the impact was weak . Mode choice had significant effects on travel perception. Compared with car trips, walking could increase positive travel perception; however, public transport trips had negative effects on travel perception. The travel perception for different transport modes affected subsequent mode choice. For travelers who had a high evaluation of travel perception during car trips, the probability of green mode  use would decrease. Travelers who gave high marks to travel perception during public transport traveling, bike trips and walking would have a higher probability to keep the transport mode use or traveling by other green modes. Information intervention had no significant effect on travel perception.The results from the process model of public transport service improvement indicate that the improvement of public transport service significantly increased the probability of public transport use. The improvement in in-vehicle time and comfort could particularly enhance public transport mode choice dramatically. In-vehicle comfort improvement had positive effects on travel perception. The relationship between mode choice and travel perception were similar to that in the information intervention process model.Policy suggestions were presented here in the light of the findings. (1) Information intervention had minor impacts on mode choice and no significant effect on travel perception. It was not suggested to only implement information intervention to change travelers’ mode choice. (2) Compared with traveling by car, walking had more positive impacts on travel perception. Promoting walking through traveler-oriented transport planning helped to enhance overall travel perception. (3) The improvement of public transport service, especially in-vehicle time reduction and in-vehicle comfort increase, helped to facilitate public transport use and increase travel perception.Future studies may focus on the following: (1) Different traveler groups may give different responses to transport interventions strategies. Travelers can be classified into different groups according to their travel characteristics and individual attributes. Then, the effects of transport interventions strategies to different traveler groups can be analyzed. (2) Different intervention strategies are often implemented simultaneously. The combination of multiple transport interventions strategies should be further considered to analyze the comprehensive impacts on mode choice and travel perception. (3) In this study, only the short-term impacts of transport interventions on mode choice and travel perception were explored. In further studies, long-term impacts should be conducted and compared to short-term impacts.


Community-based management of acute malnutrition (CMAM) is an effective program to manage children with acute malnutrition, including both severe and moderate acute malnutrition. However, little is known about continued child nutritional status after discharge from community based management of acute malnutrition programs in Ethiopia.The study aimed to identify factors associated with relapse of acute malnutrition among children 6–59 months after been discharged recovered from community based management program in South Gondar Zone, Northwest Ethiopia.A case-control study was conducted in three districts of South Gondar Zone by tracing children age 6–59 months who were reported as recovered from the community based management program. Sample size calculated for the first objective of assessing prevalence of severe acute malnutrition among children following discharge as recovery using Epi- Info version 7.1.3.3 StatCalc taking 95% CL, 17.8% post discharge relapse , 3% margin of error, design effect of 2 and adding 5% non-response rate was the largest sample size and used to this study.Children with Mid Upper Arm Circumference (MUAC) <12.5cm constituted cases and children with > = 12.5cm served as controls. Data were collected from 10 November 2017 to 30 January 2018 using a survey questionnaire and families were asked to bring children to a health facility for anthropometric measurements, following which data were entered and analyzed. Bivariate and multivariable logistic regression models were utilized to measure association between the risk factors and acute malnutrition.Overall, 1,273 participants were interviewed. The mean age in months of children was 23.1 (±9.1 SD) for cases and 23.1 (±8.9 SD) for controls. About 40% of the cases and 50% of the controls were female children. The factors associated with acute malnutrition were: male children , living in a food insecure household , non-receipt of Vitamin A supplement , prelacteal feeding , distance to water source more than 15 walk , less frequent self-reported hand washing , mother not having consumed extra food during this pregnancy/lactation , and respondent age above 30 years .The key factors contributing to relapse of acute malnutrition were related to childcare and feeding practices. Social and behavior change communication strategies targeting families at risk of undernutrition, along with improved food security and integrated programming are recommended to prevent relapse of acute malnutrition. Under-nutrition results from reductions in food intake or diet quality and is often combined with pathological causes , 2 and mChildren diagnosed with SAM are discharged recovered from the program when their weight-for-height/length is ≥–2 Z-score and they have had no oedema for at least 2 weeks or alternatively if their mid-upper-arm circumference is ≥12.5 cm and they have had no oedema for at least 2 weeks. In addition, children admitted with only bilateral pitting oedema are discharged recovered based on the anthropometric indicator routinely used in local programmes  which in(69% MAM & 9% SAM) [The CMAM program has been reported to be effective in terms of access and key performance indicators  , 14. In  9% SAM) , 27% (10 9% SAM) , and in  9% SAM) . Some st 9% SAM) , 23–25 hThe study was conducted in South Gondar Zone of Amhara region, Ethiopia. South Gondar Zone is one of the 11 administrative zones of Amhara region, Ethiopia. Debretabor is the capital city of the zone situated about 100 Km East of Bahir Dar  and 667 Km North of Addis Ababa . The Zone has 17 districts, five of which are town administrations namely Debretabor, Woreta, Mekane Eyesus, Addis Zemen, and Nefas-Mewcha. According to the Government of Ethiopia, the 2017/18 population of the Zone was 2,484,929 of which 183, 525 were children 0-4yrs old . In SoutAn unmatched case-control study was conducted from 10 November 2017 to 30 January 2018. The source populations were 6–59 months old children discharge as recovered from CMAM program in South Gondar Zone and the study populations were those children age 6–59 months following discharge as recovered in the randomly selected district of South Gondar Zone, Amhara region, Ethiopia.Case and control children were identified from those discharged as recovered after their mid upper arm circumference (MUAC) was taken and edema was assessed.Case: a child was considered as case, if he/she had MUAC <12.5cm  and/or presence of bilateral edema) after being successfully discharged recovered from CMAM program.Control: a child was considered as control, if he/she has MUAC > = 12.5cm and/or with no bilateral edema after being successfully discharged recovered from CMAM program.Children 6 to 59 months old who were discharged recovered from CMAM program.Children were excluded if their name was different from the name present in the CMAM registration logbook. In addition, children with chronic illnesses such as cerebral palsy or congenital malformations were also excluded.Sample size determination and sampling procedure. Sample size was calculated using Epi-Info version 7.1.3.3 StatCalc with 95% CL, 80% power, 1:2 case to control ratio, with child vaccination status as the main exposure (19.3% incomplete vaccination status and AOR = 1.89) [ = 1.89) , a desigRegarding the sampling procedure, a two stage sampling technique was used. Among the 17 districts of South Gondar Zone, 12 were rural administrative districts and from these rural districts, 3 districts  were selected randomly using a lottery method within which, 10 health centers were selected randomly . In the current Ethiopian health system, a health Centre has 5 cluster health posts in its catchment area and we considered 3–5 health posts per health Centre based on reported caseload, therefore 26–43 discharged recovered children were identified per health post using the one-year therapeutic multi-chart logbook (from November 2016 to October Oct 2017) as a sampling frame. Finally, through these identification procedure, parents were contacted by Health Extension Workers (HEWs) and asked to bring children to the health post for survey data collection, to measure their mid upper arm circumference and to examine them for bilateral oedema according to anthropometric standards.Children’s vaccination status was checked by looking at their immunization card and if not available, mothers were asked to recall if the child was vaccinated in as much detail as possible especially the last vaccination when they told by health professionals that the child has now completed his/her vaccination to know indirectly for measles vaccination and Bacillus Chalmette Guerin (BCG) vaccination for tuberculosis was also checked if any scar existed on a child’s arm.The outcome variable was relapse of acute malnutrition after discharged as recovered from the CMAM program in South Gondar Zone, Amhara region, Ethiopia.Independent variables included socio-economic status, demographic data, household hygiene/sanitation, awareness of recommended caring and feeding practices for children, health facility access, and household food security.The data collection tools used consisted of a checklist and questionnaire. The checklist was prepared using the stabilization Centre/outpatient therapeutic program (SC/OTP) multi-chart and registration logbook which are utilized throughout the country , 28 to iIn addition, the questions to assess HH food security status were taken from a validated questionnaire developed by Food and Nutrition Technical Assistant (FANTA) project . For furThe questionnaire was prepared in English and then translated to the local language (Amharic) and back to English to check the consistency. Mothers or primary caretakers were interviewed using the questionnaire which took approximately 20–30 minutes. The data collection took place from 10 November 2017 to 30 January 2018. A total of 15 data collectors who had SAM management training and relevant previous experience in data collection were recruited and trained for 2 days, with training content mainly focusing on anthropometric measurement techniques and on how to administer the questionnaire . The datChild MUAC was measured halfway between the olecranon and acromion process using non-stretchable tape snugly at the midpoint according to the standard and tape was checked as not pulled too tight or too loose and recorded to the nearest 0.1cm. In addition, presence of edema was assessed by grasping both feet in the hands with the thumbs on top of the feet and then pressing the thumbs gently for three seconds or a count of 101,102,103 and then releasing the thumbs. It was recorded as “0” if no pitting was detected on the feet, recorded as “+” if an indent was detected on feet, “++” if on legs and feet, and “+++” if it included the hands and face according to accepted standards .To assure data quality, data collectors and supervisors were trained for 2 consecutive days on anthropometric measurement techniques , 28 and Relapse: a child was considered as a relapsed case, if he/she had MUAC <12.5cm (both SAM and MAM) and/or presence of bilateral edema after having been successfully discharged recovered from CMAM , 13, "CaA respondent was categorized as having good hand washing practice if they reported washing hands at 3 or more of the recommended critical times/points .A household was considered to have improved drinking water if the source was either from a pipe, protected spring, protected well and/or boiled water.A respondent was considered as currently on family planning if she reported using any family planning methods currently to avoid pregnancy or extend the interval between births.Household food insecurity status was determined using the 9 item Household Food Insecurity Access Scale (HFIA) question. and Prior to assigning the food insecurity category (access), each frequency of occurrence responses was coded as 0 for all cases where the answer to the corresponding occurrence question was “no” and then the four food security categories were computed and created sequentially as recommended by FANTA . FinallyThe questionnaire was checked manually for completeness and was entered into EpiData version 3.3.2, and exported to SPSS version 20 for analysis. Data cleaning was done using frequency distribution. Anthropometric indices were generated according to the WHO’s 2006 Child Growth Standards  using WHBivariate and multivariable binary logistic regression was employed to identify predictors of acute malnutrition after children were discharged as recovered from CMAM program and those attributable variables with P value <0.05 were entered in to the final multivariate binary logistic regression model. Variables in the final model with P value <0.05 were declared statistically significant.Strength of association was determined using the odds ratio with 95% CIs. The final model was tested for model goodness of fit using Hosmer Lemeshow test. The standard error (SE) values of variables in the final model were checked for multicollinearity. Mean and standard deviation were used for continuous variables, and frequency and percentages for categorical variables when analyzing.The protocol and consent form was approved by the institutional review board (IRB) of the College of Health Sciences of Addis Ababa University, with an IRB protocol number of 068/16/SPH and meeting number of 001/2917. Written permission letters were obtained from Regional, Zonal and District Health Offices of Amhara region. Informed verbal consent was obtained from all study respondents after the purpose, risk, benefit, confidentiality, and their degree of involvement were fully explained to parents or caregivers by their local language and then data collectors signed on the consent form to indicate respondent’s agreement to start the interview. Children with MUAC <11.0cm or presence of edema were linked to OTP as this was the currently used admission criteria to therapeutic care. Nutrition education was given to all respondents after interview.rd (60%) and half (50%) of children from the cases and controls were male children. About 66% and 57% of the respondents were unable to read and write for cases and controls, respectively. About 80% and 84% respectively of the cases and the controls were farmers by occupation. From the total 445 cases, 155 were SAM and the remaining 290 were MAM cases  were interviewed providing a 97.8% overall response rate (100% for cases and 94.7% for the controls). All respondents were female, rural dwellers and Amhara in ethnicity. Respondent mean (±SD) age in years was 29.9 (±6.8) for cases and 28.9 (±6.6) for controls, respectively. Regarding child related variables, the mean age was 23.1 (± 9.1) months for the cases and 23.1(± 9.8) for the controls and also more than half (56% cases and 53% controls) were in the age range of 12–23 months. Nearly 2/3ases See Nearly two third of respondents reportedly (61% for the cases and 66% for the controls) initiated breast feeding immediately within one hour after birth. Prelacteal feeding was practiced more among the cases than the controls (9.4% vs.3.6%). More than half (51% cases and 54% controls) of respondent households’ water source was improved. About 64% of the cases and 81% of the controls were given vitamin A in the 6-month preceding the survey and also 53 to 54% of children both from the cases and controls were discharged recovered within the last 4–6 months, with an average of 5.2 months since exit from CMAM as recovered see .Variables were checked whether they were factors associated with post-discharge relapse of acute malnutrition following discharge as recovered from CMAMA program using bivariate logistic regression model and a total of 24 candidate variables with P value <0.05 were identified and entered in to the multivariate regression model to control confounding. Eleven variables retain their statistical significance when entered in to the final regression model.2  = 10.801, P. value = 0.213, indicating good model fit. In addition, multicollinearity was checked using the standard error (SE) values of variables and were in the range of 0.126 to 0.358 indicating no multicollinearity.The model goodness of fit was tested using Hosmer Lemeshow; XThe odds of relapse of acute malnutrition were 1.7 times higher among male children ; 2 times higher among those who had received pre-lacteal feeding ; 2 times higher in those residing in Ebnat district ; 1.5 times higher among children in food insecure HH , and almost 2 times higher among children not given vitamin A supplement  see In the current study, children were assessed after an average of 5.2 months of recovery and ten variables were identified as being associated with relapse of acute malnutrition The odds of relapse of acute malnutrition were higher among male than female children. This finding was in line with studies conducted in rural Southern Malawi , rural MThe odds of relapse were higher among children not given Vitamin A in the 6 months preceding the survey than among those who received Vitamin A supplement. This finding may indicate low receipt of, or access to nutrition services, such as distribution of Vitamin A capsules and was in agreement with a studies conducted in Hawassa, and Afar region, Ethiopia , 37. ThiIn the current study, children who were given pre-lacteal feeding were more acutely malnourished than those who were not. This finding affirms those in other studies conducted in India and in Dollo Ado district of Somali Region Ethiopia reporting that families which practiced pre-lacteal feeding had more acutely malnourished children –44. ThisIn the current study, place of residence (district) was associated with relapse of acute malnutrition. Topographically, Lay Gayint is one of the highland and mountainous areas in the Zone, while Ebnat is more lowland but both districts are drought-prone and rural. Children who lived in Ebnat district were more acutely malnourished compared to those who lived in Lay Gayint district. This could be due to the difference in the exposure to infectious diseases such as malaria as it is common in lowland than in highland area and children infected with malaria were found more acutely malnourished than non-exposed children . This coAs might be expected relapse of acute malnutrition after discharge as recovered was also related to HH food security status generally. The odds of relapse among children in food insecure HHs was higher than in food secure HHs. The finding was in agreement with studies conducted in India , NigeriaHaving another family member currently on SAM therapy increased the likelihood of being more acutely malnourished. In the current study, children in food-insecure households were more acutely malnourished than their counterparts, a finding supported by results of other recent studies , 53. LowThe odds of relapse were higher among children whose mother reported not consuming additional food during this pregnancy/lactation. International recommendations for pregnant and lactating women include additional daily food consumption to meet extra caloric needs . Our finIn the current study, children in a HH with access to drinking water within 15-minutes walking distance were less acutely malnourished than those whose access to drinking water was greater than 15-minutes walking distance or more. This finding is similar to that of a study done in Dollo Ado district, Somali region, Ethiopia . Those wThe World Health Organization (WHO) recommends that for optimal health outcomes families should wash hands at four key time points/critical periods: before eating, before preparing food, after defecation, and after disposal of child feces to prevent infection and malnutrition . In the Age of the index child’s mother was also a factor associated with post-discharge relapse, with children whose mother was above 30 years were being more likely to be malnourished. A study in Nepal also identified an association between acutely malnourished children and mothers aged above 35 years . This coAccording to the current CMAM protocol, the length of treatment stay at OTP is expected to be <8 weeks, otherwise considered as poor performance  but in tThe study is the first study among children successfully discharged recovered from CMAM in the study area to identify key associated factors with post-discharge relapse of acute malnutrition; such information will help program managers and nutrition focal persons to address acute malnutrition. This study however has limitations: The cases and controls were identified using MUAC only and not using both MUAC and WHZ as the two indicators could identify relapsed cases differently. The factors associated with relapse of acute malnutrition “provision of colostrum, Vitamin A supplementation, and distance from water sources” need to be interpreted cautiously as the responses “don’t know, not sure, and/or don’t remember” were classified as “no” assuming that children were not given colostrum, Vitamin A, and/or distance to water source was more than a 15minute walk. Food insecurity status could be underestimated as the data collection time was in the harvest season (October 2017 to January 2018).Several risk factors were associated with relapse of acute malnutrition following recovery . Understanding these factors may lead to the ability to predict and prevent future post discharge relapse of acute malnutrition. WASH and prelacteal feeding practices were among the drivers of optimizing child nutrition conditions following recovery, therefore, improvements in caregiving practice with mothers being the critical entry point for behavior change should be one priority interventions along with CMAM program. In addition, household food insecurity interventions through public Safety Net programs should be strengthened.S1 Data(SAV)Click here for additional data file.


Meriones libycus) were caught in their natural biotope in the Saharan desert in Algeria and showed a seasonal cycle of the testis activity, characterized by the highest peak during spring (the breeding season) and the lowest activity was registered during autumn and winter (nonbreeding season). Lhcgr mRNA expression is increased in autumn and decreased in spring. This expression varied in an opposite manner to testicular and seminal vesicle structures. The breeding periods of desert rodents should be favorable to the survival of small young, conditioned by the availability of food that occurs in the Libyan jird biotope during the long photoperiod and the high temperatures. The Libyan jird  is a wild desert rodent that is a seasonal breeder species adapted to breed when the environmental conditions can satisfy the energy and hydrous requirements of pregnant and nursing females to ensure that births occur at the most favorable time of the year. We assessed gene expression of testicular luteinizing hormone receptor (Lhcgr) correlated to testis activity. The expression of Lhcgr was evaluated using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR and the testis activity by a histological method in adult male Libyan jirds during the nonbreeding and breeding seasons. Our results showed that Lhcgr mRNA expression increased in autumn during the nonbreeding season and decreased in spring during the breeding season. This expression varied in contrast to testicular structure or function and plasma testosterone levels. These results help to elucidate this desert rodent’s seasonal sexual activity, which is correlated with central regulation.The Libyan jird ( The framework supporting the gonadotropic axis is composed of different major elements—hypothalamic gonadotropin-releasing hormone (GnRH), pituitary gonadotropins (luteinizing hormone (LH) and follicle-stimulating hormone (FSH)) and gonadal sex steroids . NumerouHence, kisspeptins, and multifunctional peptides are identified that stimulate and regulate GnRH/LH/FSH secretion at puberty and adulthood through the activation of their cognate receptor GPR54. The classical G protein-coupled receptor constitutes an important parameter in the regulation of gonadotropic axis ,4.FSHr gene is expressed by the somatic Sertoli cells that control germ cell development. LhcgR expression is restricted to the Leydig cells located in the interstitial compartment and is responsible for steroidogenesis in the testis. Data on the sexual activity associated with changes in pituitary gonadotropin expression of several wild mammal species are reported in bear [The  in bear , blue fo in bear , rams [7 in bear , and roe in bear . The Eld in bear . Photoperiod is an environmental cue, and also factors such as nutritional and social status may influence the reproductive function. Thus, variations in rainfall and temperature in dry deserts are key triggers for early reproduction in small mammals .Most rodents living in arid and semi-arid regions are seasonal breeders and show several variations in testicular activity during the year. Indeed, during the active period, the reproductive organs show considerable growth in weight; structure and ultrastructure are developed and both testicular and plasma testosterone levels increase. The nonbreeding season is typically characterized by a reduction in the size of reproductive organs  induced by a marked reduction in the testicular size and the plasma testosterone level ,13,14,15Meriones libycus), a seasonal breeder, KiSS-1 expression was higher during the breeding season compared to nonbreeding season. Whereas LHβ and FSHβ expression levels were higher during the nonbreeding season in autumn and varied in an opposite manner with testicular and seminal vesicle weights and with plasma testosterone [We previously reported that in the Libyan jird  in Bechar, Algeria. A total of 10 adult males were captured between September 1 to 10, 2010 and 2017 corresponding to the nonbreeding season, and 10 males were captured between March 10 to 20, 2010 and 2017 corresponding to the breeding season. Adult reproductive condition was evaluated according to body weight (60–140 g), and genital status was assessed during the breeding season when testis and seminal vesicle weights were higher than during other seasons. The animals were placed in individual cages . Animals were kept for approximately 24 h and then euthanized under ketamine anesthesia between 13:00 and 18:00. The left testis was removed immediately, frozen in liquid nitrogen, and stored at −80 °C until assayed, and the right testis and seminal vesicles were used for histology.The right testes and Seminal Vesicles (SV) were quickly excised, carefully freed from surrounding fat, weighed, and then fixed in 10% neutral buffered formalin for 24 h. Testes and SV were dehydrated in a series of increasing degrees of ethanol  and cleared in cyclohexane. After 12 h of impregnation in paraffin in an incubator at 60 °C, testes were embedded in paraffin. The paraffin-embedded tissue samples were sectioned at a 5 μm thickness  with a Leitz 1512 rotatory microtome . After dewaxing and rehydration, the sections were stained either with Masson’s trichroma or Azan of Heidenhain, Van Gieson’s trichroma, or Sirius red and then they were dehydrated in ethanol. The slides montage was performed with the use of a neutral resin Euckitt . The slides were observed at different magnifications  under a light microscope, and a Zeiss photomicroscope  was used to take microphotographs. For morphometric measurement, the diameter of seminiferous tubules (µm) with round or nearly round profiles were evaluated for each animal on 100 seminiferous tubules per animal, and the mean ± standard deviation (SD) of the diameter was determined by taking the average of two diameters—SI and S2 (perpendicular to one another). Height (µm) of seminal vesicle epithelial cells and height (µm) of their supranuclear zona were measured on six slides from each animal and 10 randomly selected fields from each slide were calculated. The microscope fields were examined under a 40× objective. Morphometric parameters were calibrated using manual micrometry .Total RNA was extracted from a single testis using TRIzol reagent  following the manufacturer’s instructions. The total RNA concentration was determined by spectrophotometry at 260 nm. Two micrograms were reverse transcribed in a reaction mixture (40 µl) containing 100 U of Moloney murine leukaemia virus reverse , reaction buffer , 0.2 mM deoxynucleotide triphosphates , 50 pmol oligo (dT) primers  and 20 U of ribonuclease inhibitor .Lhcgr sequences obtained from the following different species and accession numbers (Mongolian gerbil (AB571125); Syrian hamster (AB571126); rat (NM_181692.1), mouse (NM_178260.3), and opossum (NM_001144132.1), we identified the core sequence from which several sets of primers were designed to facilitate amplification of the specific polymerase chain reaction (PCR) products for Libyan jird—Lhcgr and β-actin. Primers were designed using Primer Express software ver. 3.0   . The conPCR products were purified and sequenced using a CEQ DTCS Quick Start Kit on a DNA sequencer .Lhcgr and β-actin, we designed new primers on two different exons for the quantitative real-time PCR analysis using the Primer Express software . Briefly, samples were heated for 10 min at 95 °C, followed by 40 cycles or 45 cycles of 15 s at 95 °C and then 1 min at 60°C in a total volume of 25 µl. The PCR was performed in duplicate, and a reagent blank, which was prepared using the Retro Transcriptase blank was included on each plate to detect contamination by genomic DNA. The reaction products were resolved in a 1.5% agarose gel and visualized with ethidium bromide to determine the lengths of the amplified DNA fragments. The β-actin gene was used as an endogenous control to normalize the initial RNA level. The ΔΔCt method [Lhcgr mRNA from each season compared with those of the internal standard (β-actin), and the values were compared.After identification of the specific Libyan jird cDNA sequences software . Relativt method  was usedp < 0.05 were considered to be significant.All data are presented as the mean ± standard error. Differences between means were evaluated using the Student’s t-test. We used GraphPad Prism  to determine the differences in the various parameters between seasons. Values of In the breeding season (spring), under light microscopy, the diameter of the seminiferous tubules was large (700 µm ± 8.6 µm), with a regular form and intact basement membrane a. Spermap < 0.001), and the lumen was reduced or sometimes absent (During the nonbreeding period (autumn), the diameter of seminiferous tubules decreased (300 µm ± 6.4 µm) significantly,  than that during the breeding season (0.92.10–3 ± 0.15.10–3), (p < 0.05) a change of 48%  exhibits a seasonal reproductive cycle, with a period of reproductive quiescence during autumn and winter and a period of reproductive activity during early spring until early summer [Psammomys obesus, which is in testes atrophy during spring, summer, and autumn, and the Syrian hamster, which undergoes gonadal regression in autumn and remains reproductively quiescent throughout winter, spermatogenesis is arrested at the spermatocyte stage [The adult male Libyan jird , the testis, which is in the active phase in the summer and the nonbreeding period in the spring, showed similar morphological seasonal variations [Lagostomus maximus) seminal vesicles and prostate [These morphological characteristics reflected inhibition of spermatogenic and steroidogenic activities of the testis and correlated with changes in total testicular androgens and plasma testosterone levels . In the riations . Hence, prostate ,23.Lhcgr mRNA expression increased in autumn during the nonbreeding season and decreased in spring during the breeding season. The histological results of the testes and seminal vesicles follow the same profile as the plasma testosterone previously observed [Lhcgr mRNA expression. These peripheral data will allow us to understand the regulation of reproductive function in the Libyan jird and will help to elucidate this desert rodent’s seasonal sexual activity, which is correlated with hypothalamic-pituitary regulation. Indeed, in our previous study, LHβ mRNA expression, and its cognate Lhcgr mRNA were less complex considering upregulation of both genes during the nonbreeding season and downregulation during the breeding season. The late maturational process is considered to be more dependent on LHβ expression than on KiSS-1 expression, which is apparently upregulated [LHβ and FSHβ that are opposite of their mRNA expression would be predicted by variation in KiSS-1 expression [Our results showed that observed , and theegulated . The dirpression .Lhcgr expression could be implicated in testicular kisspeptin expression. This regulation may be caused by pulsated GnRH, which remains unknown in this species. This seasonal variation of reproductive activity in a desert rodent is similar to that observed in red deer stags (Cervus elaphus). In this species, the LH levels are maximal in August during the phase of testicular redevelopment, whereas the testosterone levels are maximal from September to November, corresponding to the time of peak testicular activity and the mating season. Castrated stags had higher plasma levels of LH than those of the intact stags at all times of the year, and no clear seasonal cycle in LH levels was observed in this species [Ursus thibetanus japonicus), significant variations in plasma testosterone and inhibin concentrations and changes in FSH concentrations preceded those of these hormones with a similar tendency [FSH and LH receptor mRNAs are possibly expressed in Sertoli cells and Leydig cells, respectively, as they are in other mammals. However, neither plasma LH concentration nor testicular gonadotropin receptor mRNA expression levels varied significantly among the sampling months. These results suggest that FSH, inhibin, and testosterone have roles in testicular activity in male bears [Wang et al.  performe species . In beartendency . Hormonele bears .In adult male hedgehogs, the seasonal variations in gonadal and pituitary activities are in parallel during the year ,29. The In autumn and winter, the regular spontaneous recrudescence of pituitary/testicular activity occurs after several months of rest. This phenomenon could be a state of photo refractoriness, which is broken later in autumn, or a negative feedback control from increased levels of LH in autumn and corresponding to the development of refractoriness to short days at the central level. This hypothesis is consistent with similar results obtained in hamsters ,32.Aepyceros melampus), the increase in testosterone secretion during the hot season (activity) is associated with increased susceptibility to testicular LH receptors from an increase in circulating gonadotropins or with pituitary sensitivity to GnRH. In this species, LH is critical to start the steroidogenic activity of the testis, but for some species, testosterone and FSH are important and occur later during the maximum activity cycle to ensure spermatogenesis [In the impala (ogenesis .FSHβ and LHβ remain unknown [Psammomys obesus), a short day breeder, plasma LH concentration increases in early summer and that intravenous administration of GnRH (200 ng/100 g of body weight) fails to elicit significant season-dependent changes in LH release, whereas the increase in plasma testosterone was maximum in June–July and low between November and March–April. The summer seasonal onset of the testicular endocrine activity of the sand rat is due to increases in both in LH release and testes sensitivity to gonadotropin [LHβ messengers and therefore its secretion; conversely, weak pulsations increase transcription of FSHβ mRNA [In rams, peak LH levels are associated with the increase in testicular LH receptor concentration combined with elevated LH levels. Additionally, the peak concentrations of FSH occur later, after the LH peak, and are parallel to the testosterone cycle and sperm production. The mechanisms to explain these differences between the expressions of  unknown . GnRH ordotropin . Other sSHβ mRNA ,36.In several species, the increase in serum FSH occurs before the beginning of the breeding season and is often associated with an increase in testicular weight rather than the maintenance of spermatogenic activity ,38. In tLhcgr in the Libyan jird. On one hand, seasons can influence LHβ/Lhcgr expression, which is presumed to be essential for initiating reproductive activity during the long day in the Libyan jird. On the other hand, during the same period, high weights of gonadal tracts demonstrated testicular activity. The seasonal variation of testis activity is apparently dependent on the species and on the environmental cues that modulate the seasonal physiology to a large array of endocrine and metabolic signals.Our study demonstrates a complex interplay between the testicular activity and gonadal


R2 = 0.676, P < 0.05). Importantly, metabolic profiles were substantially different between HF patients and healthy controls. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis demonstrated that the differentially expressed metabolites are involved in signaling pathways that regulate cardiac functions. In HF patients, BNP resistance develops in association with a reduction in heart function and metabolic remodeling. It suggests possible functional roles of BNP resistance in the regulation of cardiac metabolism.Brain natriuretic peptide (BNP) is an important biological marker and regulator of cardiac function. BNP resistance is characterized by high concentrations of less functionally effective BNP and common in heart failure (HF) patients. However, the roles and consequences of BNP resistance remain poorly understood. Investigate the effects of cardiac BNP resistance and identify potential metabolic biomarkers for screening and diagnosis. Thirty patients and thirty healthy subjects were enrolled in this study. Cardiac functions were evaluated by echocardiography. The plasma levels of cyclic guanosine monophosphate (cGMP) and BNP were measured by enzyme-linked immunosorbent assay (ELISA) and the cGMP/BNP ratio is calculated to determine cardiac natriuretic peptide resistance. Liquid chromatograph tandem mass spectrometry (LC-MS) based untargeted metabolomics analysis was applied to screen metabolic changes. The cGMP/BNP ratio was markedly lower in HF patients than controls. The cGMP/BNP ratio and ejection fraction (EF) were strongly correlated ( Heart failure (HF) is a complex clinical syndrome characterized by structural disorders of ventricular filling or ejection and is unable to meet the blood supply needs of peripheral tissues , 2. ImpoBrain natriuretic peptide (BNP) is a member of the natriuretic peptide (NP) family, which plays an important role, such as natriuresis, diuresis, vasodilation, and metabolic regulation –10. All Metabolic dysregulation is one of the important components of the complex pathogenesis of HF –18. MetaIn this study, a metabolomics approach performed using liquid chromatograph tandem mass spectrometry (LC-MS/MS) was used to study the changes of metabolites in HF patients. The HF and healthy control groups were clearly distinguished by multivariate data analysis. Some metabolites were found to be significantly altered in HF patients, and the perturbed metabolic pathways were analyzed. Few studies have evaluated samples obtained from HF patients using metabolic analyses, which have shown that BNP resistance is correlated with metabolism in cardiovascular diseases. This study may, therefore, serve to identify possible metabolites that are differentially regulated between HF patients with natriuretic peptide resistance and may thereby provide a novel perspective in identifying possible metabolic biomarkers for disease screening and pathological studies. Thus, this study contributes to a better understanding of the changes that occur in cardiac energy metabolism during HF.4OAc , NH4OH , and MeOH (LC-MS) were purchased from Sigma-Aldrich . Deionized water was produced by a Milli-Q system. The chemical standards were of analytical grade with a typical purity of 99%.NHThe analyzed plasma samples were obtained from two cohorts ). The 30 HF patients were diagnosed according to the criteria of the American College of Cardiology and the American Heart Failure (ACC/AHA HF) classification system. In particular, HF patients with hypothyroidism, systemic lupus erythematosus, inflammatory disease, chronic liver disease, chronic obstructive pulmonary disease, or cancer were excluded. Blood samples were collected before treatment. HC were included as HF-free subjects with no previous history of heart failure and no history of cardiac medication. All participants were recruited from Xi'an Medical University Second Hospital from January 2017 to January 2018. As showed in Echocardiography was performed noninvasively using a 55 MHz Vevo 770 transducer  under 2.5% isoflurane gas anaesthesia. The measurements of wall thickness and left ventricular chamber diameter were obtained along the parasternal long axis in M-mode recordings of the left ventricle (LV). Then, LV fractional shortening (FS) and ejection fraction (EF) were calculated from the LV dimensions .Approximately 5 mL of blood was drawn from each subject by venipuncture and stored in anticoagulant tubes of Ethylenediaminetetraacetic acid (EDTA) after the patients' admission from the hospital. After 30 min of coagulation at room temperature, plasma was separated by centrifugation . The 400 microliters of plasma were then placed into centrifuge tubes and stored at −80°C until required.Plasma was thawed prior to the quantification of cGMP and BNP levels, which were measured in the plasma of patients and HC using commercially available ELISA kits  according to the manufacturers' instructions.µL were extracted with MeOH : Acetonitrile (ACN) . The samples were then vortexed for 30 s and sonicated for 10 min. To precipitate proteins, the samples were incubated for 1 h at −20°C followed by 15 min of centrifugation at 20,000 g and 4°C. The resulting supernatant was removed and evaporated to dryness in a vacuum concentrator. The dry extracts were then reconstituted in 40 µL/mg pro-ACN : H2O , vortexed for 30 s, and sonicated for 10 min. The extracts were centrifuged for 15 min at 20,000 rpm and 4°C to remove insoluble debris. The supernatants were then transferred to high-performance liquid chromatograph (HPLC) vials and stored at −80°C prior to LC/MS analysis.The plasma was vortexed and thawed for 30 s. To measure metabolites, sample volumes of 200 µL, and the flow rate was 0.4 ml/min. Mass Spectrometry (MS) analysis was carried out on a Q-Exactive MS/MS device in both positive and negative ion modes. (1) The relevant tuning parameters used for the probe analyses were as follows: auxiliary gas heater temperature, 400°C; sheath gas, 40; auxiliary gas, 13; and spray voltage, 3.5 kV for positive mode and negative mode. The capillary temperature was set at 350°C, and the S-lens was set at 55°C. 2. The data dependent acquisition (DDA) method was as follows: full scan range, 60 to 900 (m/z); resolution for Mass spectrometry Imaging (MSI) and ddMS2, 70,000 and 17,500, respectively; maximum injection time for MSI and ddMS2, 100 ms and 45 ms, respectively; automatic gain control (AGC) for MSI and ddMS2, 3e6 and 2e5; isolation window: 1.6 m/z; and normalized collision energies (NCE): 10, 17, 25 or 30, 40, 50. 3. The full scan method was performed as follows: full scan range: 60 to 900 (m/z); resolution: 140,000; maximum injection time: 100 ms; automatic gain control (AGC): 3e6 ions [Samples were separated on an amide column using mobile phase A consisting of water mixed with 25 mM ammonium acetate and 25 mM ammonium hydroxide and mobile phase B (ACN). The injection volume was 4 p values less than 0.05, were defined as differential metabolites [All MS raw data files (.wiff) were directly processed by the metabolomics processing software Progenesis QI . Automated baseline filtering, peak recognition, integration, retention time correction, and peak alignment were carried out to generate a multivariable data matrix containing information on the retention time, mass-to-charge ratio, and peak areas. The data matrix was normalized by the sum of the areas of the identified peaks using the MetaboAnalyst 4.0 Web service. The data were subsequently subjected to multidimensional statistical analysis using SIMCA-P 14.0 software . Principal component analysis (PCA) was first used to observe the overall feature distribution and dispersion among all groups. Orthogonal partial least-squares discriminant analysis (OPLS-DA) was performed to determine the distribution and dispersion among all groups. The metabolites with variable importance, indicated by a projection variable importance for projection (VIP) value greater than 1.0 and abolites .The free databases KEGG and mzCloud were utilized to identify the metabolites obtained in the LC-MS analysis by matching them for accurate mass, fragment ion, and retention time. Finally, commercially available standards were employed to confirm the structures of the metabolites.Initially, we consecutively recruited 30 HF patients and 30 healthy individuals as controls. The clinical characteristics of all subjects are shown in p < 0.001 for all healthy groups). Moreover, there was a good correlation between cGMP and BNP levels  samples clustered together tightly in both groups, which indicated that the excellent QC repeatability of the two groups was good and the stability of the analysis system was high. Furthermore, the retention time change, the mass error, and the peak areas of QC samples showed that there was no difference between the two groups of samples, and the experimental study was feasible, as shown in More than 1,000 molecular features were detected. 75 altered metabolites were significantly dysregulated between the HF and healthy groups, as shown in p value). The concentration tables are shown for the HF vs. healthy groups  impairment of cardiac function is correlated with BNP resistance and (b) different metabolic profiles are associated with cardiac function accompanied by BNP resistance.In this study, a total of 75 metabolites were found to significantly contribute to the difference in plasma metabolism between HF with BNP resistance group and healthy groups. Not all of these differences could be assigned as chemical identities, but the use of database matching allowed the assumption that several metabolites are recognized. Amino acids are either ingested or endogenously synthesized in the human body and play important physiological roles as basic metabolites and metabolic regulators. Amino acids are key substrates required for mitochondrial metabolism and are usually elevated in HF groups , 33. GluIn this study, we found that the healthy and HF groups had common metabolic characteristics, as characterized by increased alanine, aspartate and glutamate metabolism, glutathione metabolism, arginine and proline metabolism, vitamin B6 metabolism, arginine biosynthesis, citrate cycle (TCA cycle), purine metabolism, pyrimidine metabolism, glyoxylate, and dicarboxylate metabolism, all of which are due to increased metabolic substrates and macromolecular precursors to meet the increased energy and protein synthesis needs of cell proliferation.2 and several amino acids  and precursors of 1,4-Dihydroxy-2-naphthoyl-CoA (NADH), which are used in other important metabolic pathways.Our results suggest that the purine pathway could be associated with the progression of HF disease, and these metabolites may be developed into a biomarker analysis for HF diagnosis. Previous studies have reported that plasma uric acid is upregulated in patients with HF and is associated with HF burden. Another metabolite, adenosine, is involved in purine metabolism and has been reported to induce HF. Combined with the results of the current study, the purine metabolic pathway is strongly related to the progression of HF disease. Another important molecular group found in our results is critic acid, which is a weak acid that can be produced through the tricarboxylic acid cycle or introduced through diet. The evaluation of plasma citric acid is rarely used in the diagnosis of human diseases. In contrast, a urinary citrate excretion is a common tool for distinguishing kidney stones and renal tubular acidosis, and it plays an important role in bone diseases. The citric acid cycle, also known as the tricarboxylic acid cycle (TCA cycle) or Krebs cycle, is a series of enzyme-catalyzed chemical reactions that are vital to all aerobic organisms. The citric acid cycle also produces COWe found seven amino acids upregulated in patients with HF, whose expression levels are highly correlated with each other. Three amino acids belong to the alanine, aspartate, and glutamate metabolic pathways, which occur mainly in the endoplasmic reticulum. Amino acids constitute the basic units of all proteins and serve as a substrate for energy-providing. At the same time, they have critical roles in cell signaling, biosynthesis, transportation, and key metabolic pathways. Interestingly, our results show that in untreated HF patients, L-glutamic acid and citrate acid were upregulated. This indicates that the levels of L-glutamic acid can be used as a potential indicator for the diagnosis and treatment effect evaluation of HF disease. In addition, amino acids and glucose are the main energy substrates for cardiomyocytes to produce Adenosine triphosphate (ATP), so the upregulation of L-glutamic acid in plasma may be related to amino acid intake or metabolic dysfunction in cardiomyocytes.Taken together, the results of our study indicate that there is a unique metabolic characteristic that not only allows discrimination between HF patients and healthy groups but also allows BNP resistance with and without HF subjects, suggesting that the gradual depletion of energy reserves may be the cause of the development of cardiac impairment. Thus, our data indicate that energy metabolism is promoted in early/compensated HF states and that the depletion of metabolic capacity may lead to the progressive cardiac impairment observed in more serious cases of HF.The early detection of HF with BNP resistance would provide an opportunity to implement appropriate therapeutic strategies to prevent or decrease unfavourable disease outcomes. In the present study, we employed untargeted omics analytical strategies to comprehensively analyze which molecules are dysregulated in the plasma of HF patients, and further ELISA assays verified the reliability of our omics data. Finally, HF-related metabolic pathway identification was used in biological information. Therefore, to the best of our knowledge, this study takes the lead in provide a reliable and comprehensive view of the plasma content profile of BNP resistance in untreated HF patients. Our results provide not only potential biomarkers for evaluating the diagnosis and prognosis of HF, but also clues for gaining a better understanding of the pathological mechanisms of HF.In conclusion, we have demonstrated that metabolic disturbances occur in the plasma of HF patients with BNP resistance. We identified several dysregulated molecules, including metabolites. Our results could support the identification of novel biomarkers or effective therapeutic targets and provide clues as to upgrade the understanding of the pathology of HF. However, further validation of these candidate biomarkers in larger sample cohorts evaluated in double-blind tests is needed before they can be implemented in clinical diagnosis. In addition, the research function of biological experiments also needs to relate to illustrate the precise molecular mechanisms leading to the pathogenesis of HF.