The type of diet is very important for the maintenance of health and nutrition. How the sole source of carbohydrates from rice- or flour-based diet affect blood sugar has not been elucidated for a long time. In order to explore the effects of these diets, sixty SD rats were randomly divided into three groups: control group , rice diet group (R group), and flour diet group (F group). All the rats were fed for 7 weeks in total by the assigned diets for 4 weeks  and all by the AIN-93 diet for 3 weeks . The body weights of all the rats were monitored and serum samples were taken for testing blood glucose, biochemical indicators and untargeted lipidome. It was found that both rice and flour-based diets caused weight gain, but the flour diet had a significant increase in blood sugar and low-density lipoprotein (LDL), while a significant decrease in albumin (ALB) and triglycerides (TG). Twenty-three and 148 lipids were changed by lipidomics in the rice diet group and flour diet group, respectively, and two lipids showed the same changes in the two groups, all belonging to TGs, namely TG (16:0/16:0/16:1) and TG (16:0/16:1/18:2), which showed that a single diet source had a significant effect on the health of rats. Fortunately, we can recover this effect through the subsequent standard diet, allowing the rats to return to normal blood sugar, weight and biochemical indicators. A model can predict the diet types through the logistic regression method. Finally, we proposed that a single diet increased blood sugar and weight through a decrease in TGs, and blood sugar and weight returned to normal after a standard diet. Taken together, the short-term negative effects caused by a single diet can be recovered by a standard diet and further proves the importance of diet types. People intake different varieties of grains as the main source of carbohydrates, which may lead to different effects on health , 2, espeAt present, studies suggest that refined rice or wheat flour has adverse effects on human health, and diets based on these refined grains affect glucose tolerance . Most ofLipids are essential biomolecules that play a crucial role in human health. Driven by the significance of lipid biology, lipidomics has become an emerging technology in nutrition , especiaIn our research we chose AIN-93 diet as the feed of the control group, AIN-93 diet is the purified diet proposed by the American Institute of Nutrition. Standardization of diets for laboratory animals could reduce the variation in the whole experiment and facilitate reproducibility in research . In the 2O were all HPLC grade, obtained from Merck Company .Biochemistry analysis reagents were obtained from Hitachi Company . Leucine-enkephalin (LE) and sodium formate from Waters Company . All lipid standards and internal standards were purchased from Avanti Polar Lipids Inc. . Formic acid (≥95%), methanol, and isopropanol were all HPLC grade, obtained from Thermo Fisher Scientific . Acetonitrile and ddHHealthy male SPF-grade Sprague-Dawley (SD) rats  were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (License No. SCXK (Jing) 2018-0006). The room temperature for animal feeding was kept between 18–22°C, relative humidity between 30–70%, with 12 h of light and 12 h of darkness daily. Rats were fed with three kinds of feed based on the experimental design. All three feeds were commissioned by the company for the product. The three different feeds were AIN-93G, milled rice formula feed, and milled flour formula feed. In the latter two diets, the corn starch and sucrose in the AIN-93G diets were replaced with milled rice and refined wheat flour, respectively. Both rice and wheat flour are sourced from brands commonly found in the market. A schematic of the experimental design is shown in Total cholesterol (TCHO), triacylglycerol (TG), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) levels in serum were measured using an automatic biochemical analyzer . All assays were performed according to the manufacturer's protocols.I-Class, Waters, USA) coupled with an electrospray ionization quadrupole time-of-flight mass spectrometer . A Waters ACQUITY HSS T3 column  was used for LC separation, and the column temperature was maintained at 40°C. The MSE data were acquired in the continuum mode using the ramp collision energy in two scan functions.Lipidomics measurement was conducted according to previous studies , 22. Lipt-test. Differences were considered statistically significant at p < 0.05. Differential lipids were defined as p < 0.05 and VIP > 1.Raw data were imported into the commercial software Progenesis QI  for data processing and identification was performed using LIPIDMAPS database. Data with a relative reference deviation (RSD) of >30% in quality control (QC) samples were filtered. Partial least squares discriminant analysis (PLS-DA) was performed, and variable importance in projection (VIP) was calculated using MetaboAnalyst 5.0 . Column p < 0.05) (The results showed that rice and flour significantly increased body weight and blood glucose levels ( < 0.05) . However < 0.05) .Blood biochemical indicators could reflect the physiological characteristics of organisms. In different diets and treatment stages of rats, we found that some indicators changed significantly, including ALB, AST, LDL, TCHO, and TG, but most indicators did not change at stage 1, which indicated that the rice and flour diet did not cause significant changes in physiological and biochemical levels. Fortunately, ALB, LDL, TCHO, and TG can recover to normal at stage 2, but the difference in AST only at stage 2 . The resOver 10,000 peaks (the combination of positive and negative ion modes) were obtained through non-targeted lipidomics, and 514 quantitative and qualitative lipids were obtained through the identification and filtration of data with large deviations. Chromatograms were obtained in the positive ion mode (ES+) and negative ion mode (ES-) are shown in The PLS-DA model is a supervised multivariate statistical analysis method. We used PLS-DA for model analysis in the two experimental stages. At stage 1, there was a significant difference among the three groups. R1 and C1 were close to each other, but F1 and C1 were completely separated, and the difference was significant . At staghttps://www.lipidmaps.org/), the 514 lipids were divided into 18 categories, with the top seven categories accounting for more than 95% of the lipids; TGs accounted for approximately 50%  and TG (16:0/16:0/16:1), are the key differential lipids in the different diets administered in this study. We used these two TGs to establish a model using the logistic regression method and evaluated using the ROC curve. We obtained good AUC values of 0.8519, 1, and 0.8556 in the rice diet and AIN-93 diet, the flour diet and AIN-93 diet, and the rice and flour diets, respectively . In addiBased on the previous analysis, we clarified that different diet types affected body weights and blood glucose levels. Rice and flour diets alone can significantly increase body weights, while the flour diet alone can significantly increase blood glucose levels. The rice diet also caused a slight increase in blood glucose, but the increase was not significant . It was This study showed that using refined rice and flour as a single carbohydrate source may affect body weight, blood glucose, and blood lipid levels in rats compared with the AIN-93 diet. We accurately controlled the type and amount of diet, revealed the different effects of the carbohydrate source in the diet, and determined why different dietary types affected blood glucose and body weight through lipidomics analysis. Based on the above results, we found that two TGs can independently predict the type of diet. Although a single grain of rice and flour caused an increase in blood glucose and body weight, these parameters, including TG levels, returned to normal levels after we changed the diet to an AIN-93 diet.In the past, many studies found that dietary carbohydrate patterns could significantly change the level of blood glucose. However, most of this research compared whole grains with refined grains. Whole-grain corn powder was used to feed rats for 12 weeks in a diabetic model. The results showed that the FBG, HbAlc, TCHO, TG, LDL, and FFA contents in the experimental group were significantly reduced, glucose tolerance was significantly increased, and the degree of liver injury was reduced. These results indicated that whole grain corn powder can effectively reduce blood glucose and lipid levels . Some stLipidomics is an important branch of metabolomics that plays a remarkable role in nutrition research, especially in studying the effect of diet on health, obesity, and diabetes –33. A rePrevious studies evaluated the role of diets and were mostly performed on animals. Therefore, the potential benefits of these diets to human health should be carefully extrapolated. In our next step, we will analyze and investigate TGs and others in the people to reveal this process. The effect of bread consumption on blood glucose levels was previously studied using the gut microbiome, demonstrating a statistically significant interpersonal variability in the blood glucose response to different bread types. These results suggest that understanding dietary effects requires the integration of individual-specific factors that can also guide a personalized diet . Hence, We found that single rice and flour diets resulted in significant increases in body weights and blood glucose in rats, and this adverse effect could be recovered by the AIN-93 diet. Through lipidomics, we clarified and confirmed that two TGs were closely related to the changes in blood glucose and hence could be used as an independent factor to predict the type of diet. This study recommended that a diet with multiple carbohydrate sources could help to maintain good health.Supplementary Material, further inquiries can be directed to the corresponding authors.The original contributions presented in the study are included in the article/The animal study was reviewed and approved by the Ethics Committee of National Institute for Nutrition and Health, China CDC.ZW and XX: conceptualization. MZ, BL, and ZX: data curation. SW, HM, and JW: formal analysis. XZ: methodology. WW: visualization. SW and WW: writing—original draft. SW, WW, and XX: writing—review and editing. All authors contributed to the article and approved the submitted version.This project was funded by the Beijing Municipal Science and Technology Commission (Z191100008619006), the Chinese Nutrition Society (CNS-ZD2019090) and the Sanming Project of Medicine in Shenzhen (SZSM201611017). The funder had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.WW is employed by Beijing Junfeix technology Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Background: Traditional thermal processing is a widely used method to ensure food safety. However, thermal processing leads to a significant decline in food quality, especially in the case of fruits and vegetables. To overcome this drawback, researchers are extensively exploring alternative non-thermal High-Pressure Processing (HPP) technology to ensure microbial safety and retaining the sensory and nutritional quality of food. However, HPP is unable to inactivate the spores of some pathogenic bacteria; thus, HPP in conjunction with moderate- and low-temperature is employed for inactivating the spores of harmful microorganisms. Scope and approach: In this paper, the inactivation effect of high-pressure and high-pressure thermal processing (HPTP) on harmful microorganisms in different food systems, along with the bactericidal kinetics model followed by HPP in certain food samples, have been reviewed. In addition, the effects of different factors such as microorganism species and growth stage, process parameters and pressurization mode, and food composition on microbial inactivation under the combined high-pressure and moderate/low-temperature treatment were discussed. Key findings and conclusions: The establishment of a reliable bactericidal kinetic model and accurate prediction of microbial inactivation will be helpful for industrial design, development, and optimization of safe HPP and HPTP treatment conditions. Food safety has become a global common concern as consumers around the world are facing different degrees of food safety risks. As per the reports of the World Health Organization (WHO), approximately 600 million people  fall ill after consuming contaminated food, and approximately 420,000 die every year due to foodborne diseases .Alicyclobacillus acidoterrestris is a type of bacillus that can grow in pasteurized beverages. It has been mentioned that the use of high temperatures during heat treatment can inactivate A. acidoterrestris inoculated in apple juice, but also deteriorate the nutritional and sensory qualities of apple juice [Conventionally, several chemical and physical methods have been used for the decontamination of food material. However, these methods are associated with several drawbacks such as chemical residue in resultant food products and poor sensory and nutritional quality of food. Thus, these conventional decontamination methods were not able to meet consumers’ increasing demand for high-quality food. At present, thermal sterilization is the mainstream sterilization method in the food processing industry. Although thermal processing ensures product safety, it causes undesirable nutritional and organoleptic quality in food ,5,6. Alile juice . Among vle juice ,9. Some le juice  and muskle juice , containle juice ,12,13. THigh-pressure processing (HPP) sterilization is one of the promising techniques among recent technologies that employs low temperature and high pressure, which can kill the vast majority of harmful microorganisms in the food in just a few minutes . HPP is HPP is governed mainly by Le Chatelier’s principle, the microscopic order principle, and the Isostatic principle. Le Chatelier’s principle states that any process in equilibrium that is accompanied by a decrease in volume can be enhanced by pressure. A change in micro-order also occurs during HPP treatment, which means that the molecules will shift to a more compact structure. The Isostatic principle states that the applied pressure is instantaneously and evenly distributed within the food, independent of the structure and geometry of the food ,19. The Along with potent advantages, HPP technology also exhibits certain limitations. In general, spoilage bacteria and pathogenic microorganisms are inactivated in the pressure range of 400 to 600 MPa, but certain microorganisms exhibit strong pressure tolerance, especially Gram-positive bacteria. In addition, pressure treatments up to 1000 MPa may not completely inactivate bacterial spores at ambient temperatures ,22. It iDifferent fruit and vegetable products have different physical and biochemical characteristics, and during decontamination the different parameters of different processing methods have to be optimized to achieve optimum product quality, microbial safety, and other aspects. Therefore, much basic research is required to provide a theoretical basis for the practical application of a particular processing method. Through bactericidal kinetics, the lethal characteristics of microorganisms can be described, which plays an important role in predicting the number of microorganisms and ensuring the microbial safety of fruit and vegetable products . These kA reliable mathematical model and an accurate prediction of microbial inactivation will be helpful for the industrial design, development, and optimization of safe decontamination treatment, which in turn reduces the number of experiments ,26. TherN0 is the initial number of viable cells in the control sample (cfu/g or cfu/mL), N is the number of survivors in samples after HPP or HPTP treatment, t is the processing time, k is the inactivation rate constant of microbial number , and T,PD is the time required for one log reduction of the microbial population at a certain temperature and/or pressure.Primary models in the bactericidal kinetics of HPTP are mathematical equations that describe the changes in microbial counts induced by pressure as a function of treatment times. The first-order kinetic model is one of the common primary models employed to describe microbial log survivors in foods followed by the combination of high pressure and temperature pressure treatment ,27. The Bacillus coagulans 185A spores in tomato juice by HPTP, whereas first-order kinetics was appropriate for explaining only thermal processing [The HPP and high-pressure thermal processing (HPTP) showed a linear trend for microbial inactivation in some fruit and vegetable systems, as mentioned in ocessing .b is a rate parameter that is related to the rate at which the microorganism is inactivated, and n describes the degree of curvilinearity. When b < 1, the inactivation curve corresponds to concave-upwards (tailings). When b > 1, the inactivation curve corresponds to concave-downwards (shoulders). When b = 1, the model becomes a straight line, which is the first-order kinetic model [The Weibull model was initially used for the determination of failure time in reliability engineering and was applied to the survival curve of microorganisms, which was the combination of the accelerated failure time model and parameter distribution ,39. The ic model . This moic model .TZ-value (shown in Equation (3)) represents the temperature required to decrease D by one log cycle under a certain pressure, which can also reflect the sensitivity of microorganisms to temperature. This is equal to the reciprocal of the slope of the log D-values plotted against temperature [TZ-value, the higher the temperature sensitivity will be.D1 is the D-value at a reference temperature T1 (°C) and T is the temperature of the isothermal treatment (°C). Likewise, the PZ-value (shown in Equation (4)) is the pressure required to decrease the D value by one log cycle at a certain temperature and is equal to the reciprocal of the slope of the log D-values plotted against pressure. PZ-value can also reflect the sensitivity of microorganisms to pressure. A smaller PZ-value indicates greater sensitivity to pressure.The first-order kinetic models are established under certain temperature and pressure conditions. Unlike first-order kinetic models, the secondary models are an extension of the primary models, in which the parameters of the primary models relate to the environmental variables/conditions such as pressure or temperature . Secondaperature ,45. The Campylobacter coli and C. jejuni [C. coli and C. jejuni under the combined effects of pressure (0.1–500 MPa), temperature (10 to 65 °C), and treatment time. In addition, the Weibull model can also relate the inactivation rate parameter  to environmental variables, particularly temperature, and can be used to predict the parameter values outside the range of the variables tested [The previous studies related to high-pressure and temperature synergistic bactericidal kinetics also reflect the importance of sample temperature detection and control technology under a high-pressure environment. If the sample temperature under a high-pressure environment is not accurately detected and efficiently controlled, then the model developed to predict bacterial kinetics will be inaccurate ,46. For . jejuni . At the s tested .Y is the predicted response; B0 is a constant; iB, iiB, and ijB are model coefficients; iX and jX are the input variables ; and ε is the error term. By graphically translating RSM, operators can find the operating conditions that reduce the response, Y.Polynomial models, also known as Response Surface Methodology (RSM), are used to analyze the effect of individual factors on inactivation parameters or their interactions ,43. RSM Many factors affect the inactivation of microorganisms during HPP or HPTP. These factors include microbial species and growth stage, process parameters and pressurization method, temperature under pressure, food composition, and other factors such as food additives, water activity, and pH value ,49. in aListeria monocytogenes and Staphylococcus aureus) bacteria > fungus > Gram-negative (Escherichia coli) bacteria [Different microbe species have different sensitivities to high pressure, even within the same food system. In addition, microorganisms, including pathogens, can present significantly different responses toward high pressure. This variation exists not only between different species but also between strains of the same species . Table 2bacteria ,51,52. Hbacteria ,57,58,59B. subtilis spores were found to survive under processing conditions up to 1200 MPa at ambient temperature [Until now, inactivated bacterial spores are the biggest challenge faced by HPP during food sterilization. Bacterial spores are highly resistant compared to mold and yeast spores. These bacterial spores are likely to grow as toxin-producing cells, leading to foodborne illness and disease outbreaks. Therefore, they are often used as indicators of pasteurization and sterilization processes in foods. Conventional pressures (<600 MPa) in the food industry have difficulty inactivating the majority of spores at room temperature. Even under the extreme pressure generated by laboratory equipment, some spores can survive. For example, perature . In addiperature .The influence of pressure and holding time of HPP or HPTP on the bactericidal effect has been studied extensively . GeneralSaccharomyces cerevisiae in pineapple juice at the same holding time [The variation in pressure applied during HPP also exhibits a significant effect on microbial inactivation. The most commonly employed method is a single static high pressure (pressure holding time > 0 s). The whole high-pressure treatment process includes three parts: boost pressure, keep pressure, and relieve pressure. The boost and relief process without the keep pressure process during high-pressure treatment is called pulse-type high-pressure treatment. Different types of high-pressure pasteurization methods are composed of these two above-mentioned methods . It has ing time . Donsì eing time .Salmonella typhimurium and L. monocytogenes) with high-pressure pulses as a product of pressure and time [B. amyloliquefaciens spores [Chapleau et al. found a linear reduction in microbial count . It has been mentioned that the treatment at a moderate temperature (40–90 °C) increases the degree of protein denaturation; thus, an increase in the temperature during HPP treatment significantly enhances the lethality of microorganisms . On the Neosartorya fischeri JCM 1740 spores to high-pressure , and the control group was thermally treated at 75 °C. It was mentioned that HPP treatment  was the best method for inactivation of N. fischeri JCM 1740 ascospores in apple juice with a 3.3 log cfu/mL reduction compared to no reduction followed by thermal treatment. Similar results were also reported by Filipa et al. [A. acidoterrestris spores was observed in orange juice followed by temperature or pressure treatment alone.The spores of pathogenic microorganisms in food are extremely pressure-resistant. Thus, pressure–temperature co-treatment is considered to be one of the most effective and feasible methods for the inactivation of different pathogenic spores ,78,79. Ta et al. , that noByssochlamys nivea JCM 12,806 spores in strawberry puree. In addition, the combination of temperature and pressure was mentioned to be efficient compared to traditional thermal processing. As shown in B. coagulans 185A spores in tomato juice by high-pressure and moderate temperature synergistic process, whereas first-order kinetics were appropriate to explain microbial inactivation during thermal processing alone.However, HPP in conjunction with thermal treatment can effectively inactivate spores with a 2.7 log cfu/mL reduction at 600 MPa, 65 °C, and 10 min. Evelyn et al.  also repBys. nivea JCM 12,806 ascospores in strawberry puree at 600 MPa. Weibull model efficiently described ascospore inactivation by HPP-thermal treatment . In another study by Shao et al. [Escherichia coli K-12 count, whereas HPP at 40 °C was reported to reduce E. coli log survivors. This finding is attributed to the fact that temperature and pressure exhibit opposite effects on volume expansion. The increase in temperature contributes towards volume increase, while pressure has a reversed effect on volume. For similar reasons, some studies have also shown that for microorganisms at sublethal levels, the inactivation effect of pressure was elevated at lower temperatures [Evelyn et al.  simulateo et al. , high-preratures . In the High-pressure freezing and thawing has gained the interest of food researchers due to its inherent property to retain the quality of food. High-pressure freezing can improve the freezing effect in essence by applying pressure during freezing at atmospheric pressure. By adjusting the phase transition temperature of the water, this method increases the degree of supercooling and freezing rate during the freezing process and then changes the path of crystal nucleus formation and ice crystal growth . This meE. coli in frozen carrot juice samples was better compared to microbial inactivation in unfrozen samples  followed by high-pressure treatment at 350 MPa for 10 min at −20 °C. However, in the case of frozen samples, a better bactericidal effect (5 log cfu/mL reduction) was observed after HPP at 330 MPa for 10 min. Wang et al. [E. coli ATCC 25,922 in frozen bayberry juice after treatment at 300 MPa for less than 5 s, but the same treatment only resulted in 1.2 log cfu/mL reductions in the case of unfrozen sample. In addition, an increase in the pressure holding time resulted in a potent antibacterial effect as E. coli was not detected in the samples after treatment at 300 MPa for 5 min at −5 °C. Sami et al. [E. coli in frozen orange juice (−80 °C) after treatment at 250 MPa for 15 min. However, without freezing, the same treatment resulted only in a 0.42 log cfu/mL reduction. Moussa et al. [E. coli K-12TG1 compared to when the sample was treated at 25 °C. The above results show that the microbial inactivation by HPP treatment in frozen samples was higher compared to the unfrozen samples. In addition, below 0 °C, the inactivation effect of high-pressure treatment on tested pathogenic microorganisms was obvious.Moussa et al.  reportedg et al.  reportedi et al.  also repa et al.  reportedIn addition to the important role of crystal formation and phase transition in microbial inactivation in frozen samples during HPP, increased sensitivity of proteins to high pressure at low temperatures also leads to the rapid denaturation of proteins. That in turn plays a significant role in the reduction of the pressure resistance of microorganisms. Moreover, the structure of the cell membrane was more vulnerable to damage at low temperatures. In general, only a few studies have reported the impact of combined high-pressure and sub-zero temperatures on food sterilization. In particular, studies on the microbial inactivation kinetics of high-pressure sterilization at sub-zero (freezing) temperatures are scarce ,94. ThusA. acidoterrestris spores. The D-values were 4.17, 7.59, and 13.71 min in broths having 10, 20, and 30 Brix, respectively, showing the protective effect of the soluble solid content against the HPP-thermal process. In general, due to the complex composition of food materials, it is necessary to use the actual food materials as a medium for inoculation during high-pressure treatment. In addition, to enhance the practical application of high-pressure processing, extensive research in this field still needs to be conducted.The biochemical composition of fruits and vegetables, such as protein, carbohydrate content, and soluble solid content, significantly influence the impact of HPP or HPTP on microbial inactivation . StudiesListeria spp. and E. coli, and the results showed that the addition of the additive significantly improve the bactericidal effect of high-pressure treatment. HPP combined with natural antimicrobial compounds can effectively eliminate the pressure-resistant subpopulation and inhibit their revival or resuscitation [L. innocua and E.coli in carrot juice was achieved by the combination of HPP (300 MPa/35 °C/2 min) and natural bacteriocins, compared with less than 1 log reduction in the absence of natural bacteriocins. Zhao et al. found that HPP combined with natural bacteriocins had a significant synergistic effect on reducing the total aerobic bacteria in cucumber juice [Researchers are also using additives in the medium/food matrix to enhance the efficiency of HPP to inactivate pathogenic microorganisms or spores that were difficult to kill by HPP treatment alone. When an antibacterial agent was used as an additive, the bactericidal effect of high-pressure treatment will be enhanced significantly. Julie et al.  reportedcitation . Pokhrelcitation  found ther juice . The advE. coli was significantly weakened when the water activity of samples was reduced. Moussa et al. also reported that low water activity significantly enhanced the pressure resistance of S. cerevisiae [Water availability also affects the resistance of microbial spores towards high-pressure or high-pressure and moderate-temperature synergistic treatment . Thus, trevisiae . Due to 2O, which increases the H+ ion concentration and affect the pH variation of HPP-treated food products [The pH value of the fruits and vegetables also has a great influence on the pressure resistance of microorganisms during HPP or HPTP. Although pH alone may not be enough to inactivate microorganisms, its combination with high pressure greatly enhances treatment lethality. Microbes exposed to high-pressure processing experience irreversible damage, and cells that are not presenting any physical damage may become sensitive to the high acidity of the medium . The patproducts ,112.Presently, in the bactericidal kinetics of high-pressure or combined temperature and pressure treatments, certain empirical models are extensively used to fit the bactericidal curve. However, various factors affecting the bactericidal effect such as pH, water activity, and natural antibacterial in the medium are not considered. Although curve fitting methods such as first-order kinetics and the Weibull model are effective in evaluating experimental data, other factors such as food composition, pH value, and water activity will also significantly affect the accuracy of the model prediction when establishing the kinetic model of high-pressure sterilization. Therefore, different models are required to predict the kinetics of HPP-induced microbial inactivation and to estimate the effect of HPP under different conditions. In addition, whatever the model is, it needs to be validated in other real fruit and vegetable systems before it can be routinely used throughout the fruit and vegetable processing industry.High-pressure induced microbial inactivation is also influenced by the temperature, pH, water activity, nutritional composition of fruits and vegetables, and type of microorganism and their growth phase. Therefore, other factors besides pressure should be considered in future inactivation modelling, especially treatment temperature, which can significantly affect inactivation mechanics. Among them, the bactericidal kinetics of low temperature combined with high pressure is the least explored area, and is worthy of further study owing to its inherent advantage of producing high-quality food products. On the other hand, most of the existing studies only use one or two models to simply fit the bactericidal kinetic curve; however, these studies lack optimization, verification, evaluation, and application of the bactericidal model. That in turn results in the poor credibility of the research conclusions and applicability of the model. In addition, in the bactericidal kinetics study of high-pressure or combined temperature and pressure treatment, some empirical models are mainly applied to fit the bactericidal curve, and various factors affecting the bactericidal effect are not considered comprehensively. Therefore, it is necessary to carry out extensive research to obtain more data and an efficient bactericidal kinetic model. In addition, inactivation models need to be validated in other food systems before they can be routinely used throughout the fruit and vegetable processing industry.In addition, the high cost of high-pressure equipment and high initial investment cost is one of the important reasons that hinders the application of high-pressure technology in the food processing industry. At present, most high-pressure equipment used in the food industry has a rated pressure of ≤600 MPa. If the device operates at high pressure (>500 MPa) for a long time, the maintenance cost of the device will increase. Optimizing high-pressure production processes in food processing can save costs. Therefore, the research on high-pressure food processing technology is of great significance to the high-pressure food processing industry.


Covariate-adjusted linear regression was performed to compare the MD and healthy groups, adjusting for age, sex, and total estimated intracranial volume. We demonstrated that there were no differences in total hippocampal volume between the MD and HC groups. However, the volumes of the hippocampus-amygdala-transition-area (HATA) on the left side of the brain as well as the parasubiculum, presubiculum, and fimbria on the right side were statistically significantly smaller in the MD group than in the HC group. Furthermore, the volume of the hippocampal fissure on the right side was statistically significantly smaller in the HC group than in the MD group. In the MD group, we found a positive linear correlation between hippocampal volume and plasma BDNF concentrations in the CA4 area on the left side (p = 0.043). In contrast, in the HC group, we found a negative linear correlation between parasubiculum volume on the right side and plasma BDNF concentrations (p = 0.04). These results suggest that some hippocampal subfields may already be atrophic at the start of MD. In addition, our findings suggest that the sensitivity of the right parasubiculum region to BDNF may differ between MD and HC groups. These findings guide future research directions and, if confirmed, may ultimately inform medical guidelines.The aim of the present study was to investigate associations between hippocampal subfield volumes and plasma levels of brain-derived neurotrophic factor (BDNF) in patients experiencing a first episode of major depression (MD) ( Depression is one of the most commonly occurring mental disorders, with a total lifetime prevalence of approximately 15–18%. Depression has a significant impact on patients’ quality of life, including social connectivity, economic security, and professional attainment .A previous report from the ENIGMA study, a comprehensive investigation conducted using Magnetic Resonance Imaging (MRI) to evaluate brain structure, function, and disease, demonstrated that hippocampal and amygdala volumes were statistically significantly decreased in depressed patients as compared to healthy controls . ChronicBDNF (brain-derived neurotrophic factor), one of several neurotrophic factors that are widely present in the brain, promotes neurogenesis and synapse formation and is involved in synaptic plasticity . In streThe aim of this study was to investigate the relationship between hippocampal subregions and plasma BDNF concentrations, and to compare results between depressed patients and healthy controls. To the best of our knowledge, this is the first study to examine the relationship between hippocampal subregions and plasma BDNF levels in depressed and healthy controls.We recruited patients with major depression (MD) and healthy controls (HC) as follows. Depressed patients were recruited from among consecutively presenting patients at the University hospital of the University of Occupational and Environmental Health, Japan. All MD patients were diagnosed by board-certified psychiatry based on standard criteria delineated in the Diagnostic and Statistical Manual for Mental Disorders-5 (DSM-5). We enrolled depressed patients who had not yet started antidepressants to remove the effects of medication of these associations. The control group was recruited from a neighboring community through posting or calls. Subjects enrolled in the HC group had never been diagnosed with any psychiatric disorder according to the findings of the Structured Clinical Interview for DSM Disorders (SCID). Written informed consent was obtained from all patients prior to participation. This study was approved by the ethics review board affiliated with our hospital and was conducted in accordance with the principles of the Declaration of Helsinki.Plasma was collected from MD patients and HC in order to measure plasma BDNF; 15 mL of venous blood was collected in EDTA-2Na spray-coated blood vacutainers from each study participant, whole blood samples were centrifuged at 1,650 g for 25 min at room temperature. Subsequently, plasma samples were stored at −80°C. Plasma BDNF concentrations were measured using the BDNF Emax Immunoassay Kit , in which 96-well microplates were coated with anti-BDNF monoclonal antibody and incubated at 4°C for 18 h. The plates were incubated in a blocking buffer for 1 h at room temperature, and 100-fold diluted samples  were kept at room temperature for 2 h under conditions of horizontal shaking and were then washed with the assay buffer. The plate was also washed with the buffer. Following this, the plate was incubated with anti-human BDNF polyclonal antibody for 2 h at room temperature and was then washed with the buffer again. The plates were incubated with anti-IgY antibody conjugated to horseradish peroxidase for 1 h at room temperature to induce a color reaction with a peroxidase substrate and a tetramethylbenzidine solution. The reaction was carried out in a 1 mol/L salt solution. The reaction was stopped with 1 mol/L hydrochloric acid and the absorbance at 450 nm was measured with an Emax automated microplate reader (Promega).MRI data were acquired using a 3T MR system  equipped with an eight-channel brain phased-array coil. Images were acquired via 3D fast-spoiled gradient recalled acquisition (3D-FSPGR). The acquisition parameters were as follows: repetition time/echo time, 10/4.1 ms; flip angle, 10°; field of view, 24 cm; resolution, 0.9 × 0.9 × 1.2 mm. All images were corrected for image distortion due to gradient non-linearity using the “Grad Warp” software program . The int1 (ex vivo MRI data derived from autopsy brains (FreeSurfer open source software (v7.1.1)1  was used1 . In thisy brains . The resy brains .t-test for age, plasma BDNF levels, the estimated intracranial volume, total hippocampal volume, and each hippocampal subregion volume. Multiple regression analyses were performed to examine associations between plasma BDNF levels and hippocampal subregion volumes. For each hippocampal subregion, the correlation with plasma BDNF levels was analyzed. Covariates were adjusted for age, sex, and estimated intracranial volume. We evaluated the partial regression coefficients (β) and multiple correlation coefficients (r). In the hippocampal subregion that showed a significant correlation, the association with plasma BDNF levels was evaluated by partial correlation analysis , excluding the effects of age, sex, and estimated intracranial volume. To assess the effects of the presence or absence of depression on plasma BDNF in each hippocampal subregion, we examined multiplicative interactions between MD and BDNF in a linear regression model. P-values (two-tailed) less than 0.05 were considered statistically significant. All statistical analyses were conducted using Stata statistical software .We compared depression and HC groups using chi-square tests for sex and the independent sample p = 0.12, age: p = 0.14, plasma BDNF: p = 0.79).Thirty patients with first-episode untreated MD and 49 HC were enrolled in the current study. The main background factors are shown in t-tests are shown in p = 0.047), and there were no differences in total hippocampal volume between the MD and HC groups on either side of the brain. However, HATA volumes on the left side and parasubiculum, presubiculum, and fimbria volumes on the right side were statistically significantly smaller in the MD group as compared to the HC . Furthermore, hippocampal fissure volumes on the right side were statistically significantly smaller in the HC group as compared to the MD (p = 0.035).The comparative volumes of each hippocampal subregion between the HC and MD groups using r = 0.77, p = 0.043]. In contrast, in the HS group, we found a negative linear correlation between plasma BDNF concentrations and parasubiculum volume on the right side  . The partial regression coefficients between plasma BDNF levels and hippocampal volumes calculated for each subregion showed different distributions in the MD and HC groups (p = 0.013). = 0.04] . Both alC groups , 3. FiguIn this study, we found that the volumes of the left-sided HATA, the right-sided parasubiculum, the presubiculum, and the fimbria were statistically significantly smaller in first-episode untreated patients with MD as compared with a HC group. Results reported by Moreover, in untreated women with MD, the volumes of the left subiculum, CA2-3, CA4, dentate gyrus, and right subiculum were statistically significantly smaller than those in the HC group in prior research . We alsoBDNF is a neural-derived factor that is involved in neuroplasticity and is closely associated with neurogenesis and synaptic plasticity in the hippocampus. Prior studies conducted by this research group have reported that plasma and serum BDNF levels are statistically significantly lower in MD patients as compared with healthy controls . HoweverTo the best of our knowledge, this is the first study to examine the relationship between plasma BDNF concentrations and hippocampal subfield volumes. A prior report examining hippocampal volumes and serum BDNF concentrations in first-episode, untreated MD patients demonstrated that there was a statistically significant positive correlation between hippocampal volume and serum BDNF concentrations in MD patients, whereas there was no correlation in the HC group . A reporIn addition to the substantial strengths of this study, we acknowledge several limitations. First, we conducted a cross-sectional study enrolling a small number of cases, and thus the causality of relationship between the observed hippocampal subfield changes and plasma BDNF levels is unclear. A longitudinal study enrolling a larger number of patients is needed to evaluate these findings more thoroughly and comprehensively. Future highly powered studies should also evaluate the effects of antidepressant medications on the observed findings.Differences in hippocampal subfield volume were observed when comparing enrolled first-episode untreated patients with MD to healthy controls. Some hippocampal subfields were correlated with plasma BDNF levels, and an interaction was observed between the right parasubiculum volume in the MD and HC groups with respect to plasma BDNF levels. These results suggest that some hippocampal subfields may already be atrophic at the start of MD, and that the sensitivity of the right parasubiculum region to BDNF may differ between MD and HC groups. Our findings guide future research directions and, if confirmed, may ultimately inform medical guidelines.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.The studies involving human participants were reviewed and approved by the University of Occupational and Environmental Health, Japan. The patients/participants provided their written informed consent to participate in this study.RF, KW, NO, SK, and RY conceived and designed the experiments. RF, KW, SK, AI, NO, and RY performed the experiments. RF, KW, and NO analyzed the data. RF, KW, NO, and RY composed the manuscript. RF, KW, AI, TN, HT, YK, RI, SK, and RY provided expertise and edited the manuscript. All authors read the manuscript and are solely and jointly responsible for its content.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The COVID-19 pandemic has caused overwhelming changes in individual and community daily-life, resulting from the public health measures implemented to contain it, and also from its psychological and socio-economic consequences. These shifts and consequences impacted the entire population, but some groups are more likely to be affected by these changes, including higher education students.a) to investigate mental health status and its determinants among higher-education students in Portugal and Switzerland; and b) to explore adjustment patterns used by these students to overcome the impact of the COVID-19 pandemic.A cross-sectional study with a mixed-methods sequential explanatory design was conducted in two phases. First, an online survey was conducted among higher education students in Portugal and Switzerland, in Portuguese and French respectively. A convenience sampling method was used. Second, some participants from the first phase were invited to participate in four online focus group discussions (two in each country) using a maximum variation sampling method.n = 27), 13 adjustment strategies were found, which were subdivided into three spheres: personal, social, and contextual.The survey was answered by 1,880 students. Portuguese students revealed higher levels of stress and anxiety, but lower depression symptoms and less resilient coping compared to Swiss respondents. Hope was identified as an explanatory variable for mental health symptoms in students from both countries. In the focus groups (The results suggest that the COVID-19 pandemic had a mild to moderate impact on most of the evaluated mental health variables. Nevertheless, the students reacted and mobilized positive short-term strategies, which need to be reinforced in order to prevent long-term psychological harm. In addition, our results can inform psychosocial interventions to minimize psychological impact, anxiety, depression, and stress due to sanitary crises or other population-wide problems or disasters. COVID-19 has brought physical, psychological, and social hardship to populations around the world. Arguably, the pandemic represents a “perfect storm” for mental health due to its prolonged and unpredictable character, based on an unfamiliar and invisible danger, and its real threat to lives and livelihoods  to investigate mental health status and its determinants among higher-education students in Portugal and Switzerland; and b) to explore adjustment patterns used by students to overcome the impact of the COVID-19 pandemic.The study followed a mixed-methods approach with a cross-sectional explanatory sequential design, beginning with a quantitative phase that supplied context and participants for the subsequent qualitative phase see . AfterwaThe two parts of this international study were as follows:a) an online survey (quantitative) assessing the mental health status and psychological responses of higher-education students during the COVID-19 pandemic;b) a Focus Group study  discussing the strategies adopted by students to face the COVID-19 pandemic.Ethical approval to conduct this study was obtained from the Ethics committee in Portugal (CE/IPLEIRIA/22/2020) and Switzerland (swissethics/CER-VD-2020-02889). Informed consent was obtained from all individual participants included in the study.n = 385) was calculated considering the most conservative scenario (a proportion of 50%), a level of confidence of 95%, and an error margin of 5%. Data were collected between April 2020 and June 2021, via an online survey, sent by the institutions who participated in the study, including:This cross-sectional study involved students from Portuguese and Swiss higher-education institutions. Adults enrolled in education levels above high school, including undergraduate and graduate programs, were considered eligible. The age composition of students in this type of programs is very heterogenous, reflecting actual aging societies and lifelong learning concept. For that reason, no age limit was imposed. Students participating in Erasmus or other mobility programs were excluded. Participants were recruited using convenience sampling. Email contacts of interested participants were recorded in phase I, so they might be invited to participate in the qualitative phase. The total target population of Portuguese and Swiss higher-education students (4 Portuguese institutions and 4 Swiss institutions) was estimated at 61.000 individuals . The mina) Sociodemographic and health information: gender, age; marital status; level of studies; study area; employment (if applicable); household size; perceived health status ; and history of chronic disease.b) The Depression, Anxiety and Stress Scale-21 (DASS-21) , in Portc) The Herth Hope Index (HHI), with 12 items designed to measure hope in adults, in Portuguese  and Frend) The Brief Resilient Coping Scale (BRCS), a 4-point measure to assess an individual's tendency to cope adaptively . The Pore) The Impact of Event Scale-Revised (IES-R) , with 22Data were analyzed using SPSS Version 28. Descriptive data analysis was performed. Associations and/or differences between variables were determined with univariate and multivariate analyses. A linear regression model was used to evaluate the predictive capabilities of the independent variables with respect to the corresponding decision variables . A 0.05 level of significance was used.Phase II of this study was qualitative research to obtain additional insights into the online survey responses by carrying out Focus Group Discussions (FGDs). Before conducting the FGDs, the authors developed questions based on previous research , 26 enabThe data were coded and inductively analyzed using a thematic analysis approach . WebQDA The validity of a mixed-method study depends on both the quantitative and qualitative phases. Potential validity threats were minimized by using a large sample size in the quantitative phase, by selecting maximal variety (based on results obtained in phase I) between the participants selected for the FGDs, and by focusing equally on the validity of the two phases . To ensun = 1,528) were currently undergraduate students (Bachelor's degree), 38.4% in the field of health area, and 27.6% (n = 518) declared being employed. The majority declared no chronic illness, but 13.4% (Switzerland) and 26.3%  indicated they lived with elderly or chronically ill people.Of the 1,880 participants in the survey, 1,522 were from Portugal and 358 from Switzerland see . The meap = 0.123). A comparison of perceived health status, psychological impact, hope, and coping between the two countries is shown in p < 0.001). The overall sample had a mean depression score of 7.0 ± 5.6, mean anxiety of 5.2 ± 5.3, and mean stress of 8.2 ± 5.6. Students from Portugal reported higher levels of stress and anxiety, but lower depression levels compared to Swiss respondents .Regarding sociodemographic and health-related variables, statistically significant differences were found between the participants from both countries of origin, except for marital status .The overall mean hope score (HHI) was 35 (SD = 6.4) . The BRCS score shows that 1,129 (60.1%) participants had low, 485 (25.8%) moderate, and 263 (14%) high resilient levels of coping. Compared with the Portuguese sample, Swiss participants had significantly higher resilient coping scores, but lower global hope scores .The IES-R global mean score was 31.7 ± 19.3 . Statistically significant differences between the two groups emerged for this scale, with the Portuguese sample reporting higher psychological impact compared with the Swiss sample  had normal scores on the stress subscale, but 29.7% were in the mild range, and 6% could be classified as moderate. Regarding anxiety, 63.8% of the total sample had symptoms in the normal range; 12.5% were mild; 15.4% were moderate; 6.6% were severe; and 1.6% were extremely severe. A normal level of depressive symptoms was present in 71.5%, mild in 11.4%, moderate in 16.2%, and severe in 1% of the total sample.p < 0.001).As indicated in p < 0.001) played a protective role relative to depression. Anxiety, perception of mental health , and hope  were negatively correlated with depression in the Portuguese sample. In Swiss students, hope and overall state of health were negatively associated with anxiety, while resilience  was a positive predictor of good mental health. Hope was found to be a significant explanatory variable for all mental health symptoms, in students of both countries.As shown in p < 0.001); and all t-test values were significant at p < 0.001.In the Swiss sample, the model showed a predictive capacity ranging from 25.1% for anxiety to 49% for depression symptoms. In the Portuguese sample, explained variance ranged from 25.4% for anxiety to 44.2% for depression. Both models proved satisfactory  presentiThe four FGDs relied on a total of 27 participants . Using thematic analysis, 13 subthemes were extracted and classified into three main themes – or spheres – concerning the adjustment strategies adopted by students see .The first theme was related to the personal sphere, where students developed the most adjustment strategies in both countries. They referred to an opportunity for personal growth and adaptability. Portuguese students talked about a “window of opportunity” to be involved in academic and professional projects, although students in Switzerland did not mention this point. Focusing on work-life balance was a strategy mentioned in both countries, mostly by organizing and separating work from private life. Self-care through the adoption of healthy lifestyles was also important and was mostly related to physical activity and/or eating habits. Being compassionate with others emerged as a strategy that helped put things into perspective and remain aware of one's role in helping and protecting others, either directly through volunteer work or indirectly by staying at home and avoiding contacts. Students reported feeling overwhelmed by the quantity of information from different media and felt the need to select the most useful sources. Compliance with sanitary measures was an essential strategy that emerged in both countries with some differences, especially concerning the different waves of the pandemic.In the social sphere, students evoked new ways of communicating via digital networking to compensate for the lack of physical proximity and stay in safe contact with friends and relatives. Portuguese students mentioned new ways of being physically close, such as communicating through a window or from a balcony, while students in Switzerland found alternative ways of socializing by going for physically distanced walks with friends. A strategy that was greatly discussed in all FGs was the importance of being with family and friends. Students felt more aware of the importance of cultivating these relationships and giving value to time passed with loved ones.In the contextual sphere, students of both countries talked about the importance of adapting the “teaching/learning” environment. They found themselves in a new situation and had to adapt almost constantly to new ways of studying and learning online. Even in difficult situations, advantages were perceived by some students, such as not having to commute to the university. Tailored teaching support was a significant strategy, especially in maintaining their motivation. Portuguese students brought up that school support services helped them deal with certain difficulties, but this was not mentioned by students in Switzerland.This study set out to investigate the psychological and mental health responses of higher education students during the COVID-19 pandemic, and the coping strategies patterns they used to manage or deal with the pandemic. We found that 31.7, 44.0, and 34.1% of the participants had depression, anxiety, and stress symptoms, respectively. These rates were lower than in previous studies. For example, a meta-analysis  found a These differences may be related to when data was collected. Higher levels of depression in Switzerland and of anxiety and stress in Portugal suggest timing of data collection is relevant when investigating the prevalence of mental health symptoms. Recent findings seem to indicate that feelings of overwhelming, stress and anxiety were higher in the early months of the pandemic , 34. ThiOur results show that the COVID-19 pandemic had a worse impact on Portuguese than Swiss students. We hypothesize that this may be related to cultural differences. The Portuguese population is predominantly Catholic, with a close-knit family ethic, and experienced the pandemic as markedly disruptive, breaking family ties due to social restrictions, and highlighting rooted fatalistic values . AlthougResilience and hope feature prominently in our study, but to our knowledge they have rarely been used when investigating student reactions to the COVID-19 pandemic. Our research stresses the protective role of hope in the mental health status of students. Similar results have highlighted the protective benefits of hope on cognitive abilities in uncertain times , 39 and Our qualitative findings in both countries showed that students developed adjustment strategies mostly in the personal sphere. Confinement and isolation may have made students more introspective, leading to solitude and, in turn, to the development of such strategies. As shown by Höglinger and Heiniger , feelingStudents reported using social media and technology to maintain contact with family members and find information, an important coping strategy to deal with the pandemic lockdown found on other studies , 42, 44.The findings of this study suggest that the COVID-19 pandemic had a mild to moderate impact concerning most of the evaluated mental health variables. Nevertheless, the students reacted and mobilized positive short-term strategies that should be reinforced in order to protect them from long-term psychological harm. Furthermore, these strategies help students to stay mentally healthy and mitigate delayed-onset post-traumatic stress disorder during the COVID-19 pandemic .At a practical level, we suggest that higher education institutions reorganize teaching methods that provide greater consideration for student psychosocial functioning. Goal 3 of the 2030 Agenda for Sustainable Development  underlinAt a research level, it is necessary to anticipate future crises and prepare for the transition to the post-COVID-19 world. We need to understand the implications of pandemics on the development of the academic, professional, and personal paths of students. Generally, our understanding of student health must integrate knowledge of their pre-existing health conditions. Future research would benefit from a prospective study with a comparison group that would help understand the longer-term impact of epidemics on the development of psychopathological symptoms. Also, studies investigating longitudinal data on risk and protective factors  would be helpful.At a policy level, we believe countries should introduce new forms of mental health support, including information materials (mostly online) and mental health support phone lines. It may be worthwhile to shift some mental health services to a telemedicine format and increase investment in mental health . MoreoveOur results provide a theoretical basis for a broad audience of educators and researchers and may add to a better understanding of the impact of COVID-19 on higher education. Another strength is the mixed-methods research design and the use of valid and reliable instruments. The multicentric online survey provided broad ecological validity, while the focus groups brought explorative depth and consistency, and raised factors that could potentially mitigate some of the negative impacts. Despite these strengths, this study has some limitations. First, data collection did not occur at the same time in each country, because each was submitted to separate ethics committees that did not operate under the same rules or at the same speed. Second, there is an absence of information on pre-existing mental health conditions, which we suggest should be included in future research on the topic. Third, the use of non-probability sampling procedures and self-reporting measures limit the representativeness of the study. Fourth, the sample lacks homogeneous criteria since most of the participants were female with Portuguese nationality. Lastly, our conclusions are based on cross-sectional data, so we are not able to report on changes over time and establish causality.Our findings support the idea that the COVID-19 pandemic has a significant impact on the mental health of higher education students. In the future, it would be desirable to focus on positive ways of coping and the potential of hope in this regard. Based on our results, positive coping and hope should be integrated into the standard training of students across all study areas. Indeed, a greater focus on protective factors may be a promising way to prepare students – and others – for future hardships, at individual, community, and societal levels.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Ethics Committee in Portugal (CE/IPLEIRIA/22/2020) and Switzerland (swissethics/CER-VD-2020-02889). The patients/participants provided their written informed consent to participate in this study.The study was conceived and designed by AQ, MD, and CL in Portugal and by FS-M and TC-N in Switzerland. Jointly, all five set up an international collaboration, completed the study plan and obtained funding. Data collection and data analysis were carried out by DA, FS-M, and TC-N in Switzerland and by AQ, CL, MD, and ZC in Portugal. DC and CL drafted the first version of the manuscript. All authors contributed to the article and approved the submitted version.This work is funded by CCISP-HES.SO Collaborative Research, through: for Portugal, FCT and Fundação para a Ciência e a Tecnologia, I.P. (UIDB/05704/2020 and UIDP/05704/2020) and under the Scientific Employment Stimulus-Institutional Call—[CEECINST/00051/2018]; for Switzerland, HES-SO Rectorat-Impulsions. Call Swiss-Portuguese Projects (103744/R-PORTUG20-08). Open access funding was provided by the University of Geneva.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Coronavirus is a life-threatening disease with many devastating psychological, emotional, social, and sexual implications, especially for vulnerable people. This cross-sectional study aimed to assess adolescent health and anxiety during the COVID-19 pandemic. 1300 male adolescents filled out the Male Adolescent Health Need Assessment Scale (MAHNAS) and corona-related anxiety scale (CRAS) from January to May 2021. Data analysis was done using SPSS version 22. The mean age of adolescents was 15.5±2.25, and most of them (65%) were studying in high school. The mean scores for physical health were 78±7.55, psychological health 48.8±5.55, social health 48.21±8.61, sexual health 50.35±9.05, and anxiety scores were severe (46.76) in this study. There was a significant negative relationship between anxiety and health needs. The COVID-19 pandemic greatly affected the health needs of adolescents. Effective approaches and policies in this crisis, especially for adolescents, can alleviate the anxious reactions and be a way to meet their health needs appropriately. The World Health Organization (WHO) announced a health status crisis of international concern in 2020. SARS-CoV-2 epidemic is becoming a global problem , 2. InhaThere is no special treatment so far. The logical way to cope with SARS-CoV-2 is by controlling the source of infection, guarding vulnerable people, and cutting off the transmission. People should take measures to protect themselves, such as home quarantine, social contacts limitation, and use a mask in public places . In addiAccording to the WHO declaration, this universal situation may negatively impact the well-being of individuals, which in turn impacts individuals' performance . PreviouOne of the most vulnerable age groups who suffered greatly from this crisis is adolescents. The period between childhood and adulthood, from ages 10 to 19, has been identified as adolescence. It is a unique phase of human development and a unique time for laying the foundations of good health .However, the COVID-19 pandemic is putting the health and well-fare of all children and adolescents in danger. Injustice in this field is increasing every day, and there are no suitable health interventions for adolescents, adolescents being a vulnerable segment of society. Schools were closed in many countries at the height of lockdown, affecting 1.6 billion students . The aimThis study had a cross-sectional correlational design and recruited 1300 male adolescents (13–19 y/o) studying in high schools from January to May 2021. Inclusion criteria were not being married and studying in public schools. The exclusion criteria were declining to participate after completing the questionnaire. The sampling method was random multistage, and we used the table of random numbers with Tehran schools. The number of samples based on the proportion of adolescents living in each area was estimated until the total sample was completed. Online classes were also randomly selected in each school, and students were requested to fill out data collection tools online or offline according to their preferences. The demographic questionnaire and the Male Adolescent Health Need Assessment Scale (MAHNAS) were used. The MAHNAS consisted of 49 questions under the four dimensions of “sexual health”, “physical health”, “psychological health”, and “social health” with a 5-point Likert type scale . The scores obtained ranged from 49 to 245, with higher scores indicating a better health condition and lower needs for health supervision. The psychometric properties of this scale were controlled using Cronbach's α coefficient of .79 and intraclass correlation index of .89 using a test-retest method . The corThe mean age and standard deviation of 1300 male adolescents in this study were 15.5±2.25 years. Most of them were in high school (65%), had mothers with diplomas (41.2%), and fathers with bachelor's degrees (37.8%). Most mothers were employed (33%), and fathers were business owners (41.9%). According to the Kolmogorov Smirnov test, the data had normal distribution, and the large sample size allowed us to use parametric statistical tests. This study aimed to evaluate the health needs of male adolescents during the COVID-19 pandemic. Most research on Covid-19 has been focused on adults and less on adolescents. This study reported a high prevalence of mental health problems in adolescents. Moreover, valid and reliable tools used to assess adolescents' health need status during the Covid-19 crisis can identify the real health needs of adolescents and inform policy-making. The large sample size and sampling of various parts of the city and including various health domain needs were other positive hallmarks of this study.According to the results, the health needs of the physical dimension were favorable, and this may be since, during the Covid-19 crisis, individuals were more observant of their private and public health, such as washing hands, wearing masks, and forced distance. Proper nutritional status, especially to strengthen the immune system during the Covid-19, was another measure of good physical health status in this study. Furthermore, in a previous study in Iran, the state of physical health was also estimated to be in good condition .One study in Ireland evaluated the physical activity of adolescents during the Covid-19 pandemic and declared that adolescents reported less physical activity (50%), no change (30%), or did more physical activity during lockdown (20%). Adolescents who did less physical activity were more likely to be overweight .On the other hand, many social and psychological health needs were evident during the Covid-19 crisis among Iranian adolescents. In fact, most of the health needs in this area were reported. The Covid-19 crisis has increased these needs, and mental health services and interventions should be planned in public and private institutions to reduce the crisis in adolescents. The COVID-19 outburst triggered difficulties for child psychiatry and mental health services that must be directed, including public institutions covering interventions for major public health crises affecting youth. A greater challenge for this age group is dealing with lockdown and quarantine, which may push children into calamity and weaken families, especially when severe quarantine exists in the mentally or psychologically vulnerable . Young pMany studies indicated that restrictions and limitations in previous pandemics and epidemics put pressure, stress, anxiety, emotional fatigue, post-traumatic stress disorder, and fear on the human being. Adolescents experience other severe stressors during a pandemic lockdown and quarantine, such as stress, infection, death, imprisonment, social contact limitation, helplessness, depression, anxiety, panic attacks, and even suicidality –19.Several investigations in China declared that the rate of depression, anxiety, and psychological stress during COVID-19 was higher in children and adolescents –22. SomeSocial factors in the health of adolescents have always played an undeniable role. Adolescents' health generally refers to physical health and neglected social health. Epidemiological transmission changed the profile of adolescents' health needs. Lack of knowledge and attention to adolescents' health needs leads to high-risk situations, increased addiction rates in adolescents, academic failure, and psychosocial disorders. Adolescent mortality is not common; instead, the global burden of disease and disability is greater. Adolescent health crises need to be addressed and prevented on time . AccordiAnother aspect of health needs is sexual health, which was not in a good or bad situation. According to the literature, sexual education has been less observed in the real context. Half of the adolescents' sexual problems are ignored in Iran , 28. TheUnfortunately, children are out of control of the health system from the age of eight and do not enter until middle age obligatory. Many researchers similarly believed that sexual health policies needed to be revised. Considering appropriate ideas, interests, strategies, and tactics to perform evidence-based health interventions in sexual and reproductive health is mandatory , 31.In this study, coronavirus anxiety affected all aspects of teenage health. Obviously, fears concerning health are a typical response in the face of the Covid-19 pandemic. Children and young people are often exposed to death messages from the media and authorities and Covid-19 will probably be the first major global health threat to young people during the pandemic. This fear jeopardizes adolescents' and children's beliefs that the world is a safe place to live . TherefoIn our study, all domains of adolescent health were affected by the Covid-19 except for physical health. Therefore, more attention and policies are needed to meet the needs of adolescents in Iran.


A percentage of these cells survive in maternal tissues for decades generating a population of fetal microchimeric cells (fMCs), whose biological role is unclear. The aim of this study was to investigate the association between the sex of offspring, an indirect marker of fMCs, and magnetic resonance imaging (MRI) features in women with multiple sclerosis (MS).During pregnancy, fetal cells can migrate to the mother We recruited 26 nulliparous MS patients (NPp), 20 patients with at least one male son (XYp), and 8 patients with only daughters (XXp). Each patient underwent brain MR scan to acquire 3D-T2w FLAIR FatSat and 3D-T1w FSPGR/TFE. Lesion Segmentation Tool (LST) and FreeSurfer were used to obtain quantitative data from MRI. Additional data were collected using medical records. Multiple regression models were applied to evaluate the association between sex of offspring and MS data.p = 0.022), smaller left entorhinal volume , and lower thickness in the following cortical areas: left paracentral , left precuneus , right lateral occipital . NPp also had lower thickness in left paracentral cortex , left precalcarine cortex , and right paracentral cortex  when compared to XYp. Comparing XYp and XXp, we found that XYp had higher thickness in left cuneus  and left pericalcarine areas  and lower thickness in right lateral occipital cortex .Comparing NPp and XXp, we found that NPp had larger 4th ventricle volume (2.02 ± 0.59 vs. 1.70 ± 0.41; Our findings suggested an association between the sex of offspring and brain atrophy. Considering the sex of offspring as an indirect marker of fMCs, we speculated that fMCs could accumulate in different brain areas modulating MS neuropathological processes. Multiple Sclerosis (MS) is a chronic, disabling, autoimmune disorder of the central nervous system (CNS) characterized by both inflammatory and degenerative processes which result in demyelination and axonal damage. Immunological, histopathological, genetic, and therapeutic evidence suggests the autoimmune origin of MS, nonetheless the etiopathogenetic mechanism which triggers the autoimmune reaction and sustains the neurodegeneration has still to be uncovered.The incidence and prevalence of the disease among women is at least twice the figures for men, with a ratio of women to men that varies throughout geographical areas . HoweverOver the last 40 years, several authors have confirmed the existence of persisting fetal cells  in maternal blood and other tissues, demonstrating that pregnancy may establish a long-term, low-grade chimeric state in the women . Living The available data concerning the role of the microchimerism in MS are still limited. Pregnancy is considered to be a protective factor in MS, an effect likely due to the modulation of the immune system and to possible changes in the brain during pregnancy . HoweverThe aim of our retrospective, observational study was to investigate the impact of microchimerism on the neurodegenerative and inflammatory processes that characterize the disease evaluating the magnetic resonance (MR) patterns of MS female patients without a history of pregnancy, with at least a male son, and with only female daughters.In a Fifty-four consecutive patients attending the Center for the Diagnosis and Treatment of Multiple Sclerosis and Demyelinating Disorders of the University Hospital “Paolo Giaccone” of Palermo, Italy and fulfilling the diagnosis of MS according to the revised McDonald criteria  were ret1 for SPM. 3D T1w pulse sequences were analyzed to obtain data on whole-brain and regional volumes using FreeSurfer software suite.2 MR scanner was included as a categorial covariate in the statistical analysis. LST toolbox and FreeSurfer software were used to obtain quantitative data on white matter, cortical, and subcortical areas. We applied the Aseg, Aparc, Wmparc Free Surfer Atlases to obtain information on regional volumes.Each patient had undergone a 1.5 T magnetic resonance (MR) brain scan at the Neuroradiology Unit of the University Hospital “Paolo Giaccone” of Palermo. The following MR scanners were used: Signa HDxt—GE Medical System, Milwaukee, WI, USA and Achieva—Philips Medical Systems, Cleveland, OH, USA; both systems were equipped with an 8-channel phased-array head coil. The imaging protocol included three-dimensional (3D) T2-weighted (T2w) fluid attenuated inversion recovery (FLAIR) with fat saturation (Fat Sat) and 3D T1-weighted (T1w) fast spoiled gradient echo (FSPGR) pulse sequences or 3D Turbo Field Echo (TFE) pulse sequences on the GE and on the Philips scanner, respectively  demographic information, (b) prevalence of comorbidity, including every type of comorbidity and specifically other autoimmune (AI) comorbidity, (c) family history for MS or other AI diseases, (d) clinical data at onset, (e) clinical, neuroradiological, and paraclinical data at diagnosis, and (f) clinical features at recruitment.The study was conducted in accordance with the International Conference on Harmonization guidelines for Good Clinical Practice and the Declaration of Helsinki. All patients gave informed consent upon data collection and analysis.Statistical analysis was performed using STATA software version 17.0 (StataCorp LLC 2021). Demographic, neuroradiological, clinical, and paraclinical characteristics of patients were summarized through counts and percentages for categorical variables. Quantitative variables were reported as mean ± standard deviation (SD) and median and interquartile range (IQR) when the variable distribution was asymmetric.The analysis of neuroradiological data was performed considering the pregnancy status at the MRI (NPp vs. XXp vs. XYp at recruitment). We built a linear regression model with adjustments for age, disease duration, and MRI scan to evaluate the relationship between the sex of offspring and disease features at recruitment. Additional adjustments for number of previous DMTs and number of previous pregnancies were performed where applicable.Data at disease onset and diagnosis were analyzed according to the pregnancy status of patients before disease onset (NPp vs. XXp vs. XYp at onset). A linear regression model with adjustments for age at onset were applied to evaluate the association between the sex of offspring and disease features at onset and diagnosis.p-value = 0.014; 95% CI = 1.897 to 16.295) and XYp . We did not find any difference comparing the prevalence of comorbidity and family history for MS or other autoimmune diseases between the three groups , while the time-gap between pregnancy and onset was similar in the two groups   .p-value < 0.001; 95% CI = 5.816 to 16.335)  vs. 2/11 (18.18%); coeff = −0.374; p-value = 0.029; 95% CI = −0.709 to −0.039]  and XYp  and that the percentage of patients with an Expanded Disability Status Scale (EDSS) score ≥ 3.0 was higher in NPp when compared with XYp   . Compari −0.039] . At last −0.026] .Main findings for whole-brain and regional gray and white matter volumes are reported in p-value = 0.022; 95% CI = −0.708 to −0.057) and the right globus pallidus was smaller in NPp group .Analyzing the subcortical structures, the 4th ventricle was larger in NPp group when compared with XYp group  and XYp . The same group had a lower thickness in comparison to XXp in left precuneus  and right lateral occipital cortex . NPp also had a lower thickness in comparison to XYp in left precalcarine  and right paracentral areas . Comparing the cortical thickness in patients with at least a child, we found that XYp had a higher thickness in the left cuneus  and in the left pericalcarine cortex  and a lower thickness in right lateral occipital cortex .In the cortical areas, NPp had a lower thickness in the left paracentral cortex when compared with XXp .Finally, the analysis of white matter volumes demonstrated that the left entorhinal volume was lower in NPp (0.55 ± 0.17) when compared with XXp  NPp were supposed not to have fMCs, (2) XXp were supposed to carry only XX fMCs (XXp), and (3) XYp were supposed to be carriers of XY fMCs. The data showed that NPp had lower brain volumes in several cortical areas, as well as in some subcortical and white matter volumes. More specifically, comparing NPp and XXp, we found that the former group had a larger 4th ventricle and smaller right pallidum and left entorhinal volumes. NPp also reported a lower thickness in left paracentral cortex, left precuneus cortex, and right lateral occipital cortex when compared with XXp. A similar trend was observed comparing NPp and XYp: NPp group had a lower thickness in left paracentral cortex, left precalcarine cortex, and right paracentral cortex. Interestingly, at the comparison between XYp and XXp, we observed that the thickness was higher in XYp in the left cuneus cortex and left pericalcarine cortex, while XXp had a higher thickness in the right lateral occipital cortex.Pregnancy is considered to be protective in MS , and intIn A possible role of fMCs in autoimmune diseases has been hypothesized , but theIn The data reported in this study showed that women with at least a previous pregnancy had higher regional brain volumes and thickness. Several pregnancy-associated changes, including circulating hormones and epigenetic changes, have been previously suggested as protective variables in pregnancy and it is possible to speculate that the same biological mechanisms could influence the neuroradiological outcomes . HoweverProceeding from the available literature and the findings reported in this study, we speculated that these fMCs could act a neuroprotective role, homing in gray matter and white matter areas, activating processes of neurogenesis and circuit integration, and modulating the neurodegeneration associated to MS. It is worth to note that most of the differences were observed in cortical areas: in The retrospective selection of patients and the subsequently lack of blood samples at MRI acquisition for the direct research of fMCs constitute the main limitations of our study. To overcome this potential bias, data were collected from official medical records and information about pregnancies and abortions were obtained directly from the patients. Therefore, the available literature on acquisition of microchimeric cells during pregnancy and the high accuracy in data collection support our classification of patients. A second limitation is linked to the unbalanced numerosity between the three groups, which could have reduced the statistical power in some analysis and covered additional differences. Moving from these hypothesis-generating results, we designed a prospective, longitudinal study on a larger population of patients, which is still in progress.In conclusion, to our knowledge, this is the first study investigating the neuroradiological differences in MS patients according to the sex of offspring as an indirect marker of fMCs. The main limitation of the study is related to the lack of dosage of fMCs in patient blood sample. Nonetheless, our findings suggested that having a male or a female son is associated to different MRI features in some areas. Further studies are required to determine whether this phenomenon could be related to feto-maternal microchimerism and the role of fMCs in neurodegenerative diseases.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.Ethical approval was not provided for this study on human participants because data for this project were collected from clinical practice. The patients/participants provided their written informed consent to participate in this study.AB and PR have made a substantial contribution to the concept, design, and conduction of the study and drafted the article or revised it critically for important intellectual content. AB have contributed to the statistical analysis of the data. AB, MaA, GSc, ClG, SI, and MiA have contributed to the data collection for the article. CeG, MMa, GL, MMi, and IG have collected and processed the MRI data for this project. All authors approved the version to be published.