Rhinovirus being the most prevalent . Other viruses, such as Influenza A/B, Metapneumovirus, and Adenovirus, also showed increased prevalence. Disease severity and mean duration of hospitalization were similar between the two seasons. Conclusions: Our study highlights increased prevalence in respiratory viruses, including RSV and Rhinovirus, following the easing of NPIs. The prevalence in respiratory viruses, including RSV and Rhinovirus, increased in the second season compared to the first one. Interestingly, RSV’s peak incidence shifted from February to November. The emergence of rhinovirus as the most prevalent respiratory virus during certain months suggests viral competition and dynamic changes in viral circulation. The overall severity of respiratory infections remained relatively stable between the seasons.Background: The COVID-19 pandemic has had a significant impact on the epidemiology of respiratory viruses. Non-pharmaceutical interventions (NPIs) led to a dramatic reduction in respiratory infections. However, the long-term effects on respiratory virus epidemiology remain unclear. Materials and Methods: We conducted a comparative study on hospitalized pediatric patients with respiratory illness during two seasons: 1 October 2021 to 15 March 2022 and 1 October 2022 to 15 March 2023. We compared the type of virus, mean duration of hospitalization, and disease severity. Results: In the first season, 47.1% of patients (65/138) tested positive for at least one respiratory virus, with respiratory syncytial virus (RSV) being the most frequent (23.2%). In the second season, 82.9% of patients (102/123) tested positive, with RSV and  The global experience of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has significantly impacted the epidemiology of respiratory viruses. In 2020, implementing non-pharmaceutical interventions (NPIs), such as hand hygiene measures, social distancing, and face masks, led to a dramatic reduction in respiratory infections ,10,11,12All children consecutively hospitalized due to respiratory illness at the Department of Clinical and Experimental Medicine, Pediatric Respiratory Unit, San Marco Hospital, University of Catania, were included in this study. The study period spanned from 1 October 2021 to 15 March 2022 and from 1 October 2022 to 15 March 2023. Respiratory virus testing was performed using a nasal swab on all included patients. The comparison between the two seasons focused on the following endpoints: viral agent, mean duration of hospitalization, and disease severity assessed by clinical respiratory score (CRS) [t-test for unpaired data was applied to compare the mean duration of hospitalization, and a chi-square test was applied to relate the severity of viruses.For statistical analysis, SPSS 28.0.1.1 software was used. A 2-tailed Over the period 1 October 2021 to 15 March 2022, among the 138 patients admitted to our department for respiratory illness, 47.1% (65/138 patients) resulted positive for at least one respiratory virus: RSV was the most frequent one (23.2%), followed by Bocavirus (10.87%), Rhinovirus (RV) (4.3%), Parainfluenza virus (2.9%), Enterovirus (2.17%), endemic Coronavirus (1.45%), and Metapneumovirus (1.45%). A total of 53.6% of patients were negative.Over the period 1 October 2022 to 15 March 2023, among the 123 patients admitted to our department for respiratory illness, 102 resulted positive for at least one respiratory virus: RSV was the most frequent one (28.38%), followed by Rhinovirus (27.03%), Influenza A/B virus (12.16%), Metapneumovirus (6.76%), Adenovirus (5.41%), endemic Coronavirus (2.70%), and Parainfluenza virus (2.70%). No Bocaviruses were detected. A total of 14.29% of patients were negative. In examining the severity, in 2021/2022, 105 patients had mild CRS values, 24 patients had moderate values, and 9 patients had severe values. In 2022/2023, 99 patients had mild CRS values, 15 patients had moderate values, and 9 patients had severe values. As shown in Influenza virus in a community among kindergarteners from September to December [Influenza virus infection than RSV, although the latter caused the most severe cases. Notably, they observed a significant proportion of patients with RSV infection who missed their first exposure. Furthermore, when comparing previous seasons, Campo and Redlberger-Fritz noted an increased number of Influenza C virus cases in Austria during 2022, and they concluded that this could likely be attributed to a decreased prevalence in prior seasons due to non-pharmaceutical interventions (NPIs) [As suggested by the European Respiratory Society (ERS) in 2023, the emergence of SARS-CoV-2 has underscored the importance of respiratory surveillance for respiratory viruses, thereby shedding light on the lessons learned from the COVID-19 pandemic . RespiraDecember . SimilarDecember , in comps (NPIs) . p < 0.05) in 2022–2023. This discrepancy suggested a significant shift in viral activity within the population after lockdown. The lack of immune stimulation in children and adults could explain it. During 2022–2023, RSV emerged as the most prevalent virus (28.38%), confirming the trend already observed in the previous year (p > 0.05). In the previous year, RSV emerged as the predominant respiratory virus (23.2% of patients). In the current year, we witnessed a drastic and noteworthy surge in the circulation of other respiratory viruses. Compared to the previous year, a significantly higher prevalence in other respiratory viruses was observed (p < 0.05): RV, Influenza A and B, Adenovirus, and Metapneumovirus. In contrast, no cases of Bocavirus were reported, which showed a statistically significant difference from the previous year (p < 0.05). RSV, endemic Coronaviruses, and Parainfluenza viruses showed no statistical differences (p > 0.05). Another intriguing finding from our study was the observed peak incidence in RSV, which traditionally occurred around February before the pandemic. However, post-pandemic, the peak incidence appeared to shift to November. The changes in the seasonal pattern of RSV raised questions about the underlying factors influencing its circulation and highlighted the need for further investigation. Notably, in the previous epidemic, RSV exhibited dominance throughout the entire epidemic phase. However, an intriguing phenomenon unfolded this year, possibly attributable to viral competition. As shown in In our comprehensive epidemiological surveillance study focusing on respiratory viruses in children, we made several significant observations that shed light on the dynamics of viral infections during the studied periods. Our findings revealed a notable increase in the prevalence in respiratory viruses this year compared to previous years. Specifically, while 74 patients (53.26%) tested negative for viruses in 2021–2022, that figure decreased to only 22 patients (14.86) (p = 0.73). Regarding the CRS, disease severity differences were not significant (p = 0.49). These findings confirm the crucial role of robust primary prevention strategies in counteracting respiratory infections in children. The need for effective measures, such as vaccination programs, hygiene practices, and public health interventions, becomes even more pronounced in light of the evolving viral epidemiology. Moreover, it is vital to emphasize the significance of ongoing epidemiological surveillance in the post-pandemic era. The shifts observed in viral prevalence and the patterns of respiratory infections highlight the dynamic nature of viral circulation and the need for vigilant monitoring. Understanding and replying to these epidemiological changes are essential to improve public health strategies, allocate resources effectively, and develop targeted interventions for adequate disease prevention and control.Several respiratory viruses can circulate during the same period and can simultaneously or sequentially infect the respiratory tract, leading to virus–virus interactions. At the host level, the course of a virus infection could be influenced by a previous or concurrent infection by another virus. Infection by a first virus could potentiate or decrease the infection and replication of a second virus, resulting in a positive (additive or synergistic) or negative (antagonistic) interaction . OutsideInfluenza A and B, Adenovirus, and Metapneumovirus, compared to the previous year. We highlighted an intriguing phenomenon that occurred this year, potentially attributed to viral competition: in January, RSV abruptly disappeared and RV prevalence substantially increased, making RV the most prevalent respiratory virus during January and February. Despite the increased circulation of respiratory viruses, our study did not reveal significant differences in the severity of respiratory infections between the current and previous epidemics. These data suggest that the overall severity of infections remained relatively unchanged. These findings underscore the importance of robust primary prevention strategies in counteracting respiratory infections in children. Vaccination programs, hygiene practices, and public health interventions are crucial, especially in light of the evolving viral epidemiology. The observed shifts in viral prevalence and infection patterns highlight the dynamic nature of viral circulation and the need for vigilant monitoring. Our comprehensive epidemiological surveillance study highlighted several noteworthy observations about viral dynamics during the investigated periods. In 2022–2023, a remarkable increase in the prevalence in respiratory viruses compared to previous years was reported, suggesting a significant change in viral activity within the population, possibly due to decreased immune stimulation. RSV emerged as the most prevalent virus during the 2022–2023 period, confirming the trend observed in the previous year. Additionally, there was a significant surge in the circulation of other respiratory viruses, including human RV,


The costs of treating an elderly patient with gout are largely related to the treatment of concomitant pathological conditions and complications. Determining the costs of treating the disease is made by clinical and economic analysis, the task of which is to calculate the “cost of the illness”.A descriptive, selective study of 237 patients with gout. In order to study the clinical characteristics of gout in the elderly, all those included in the study was divided according to age at the time of examination: group I made up of patients with gout up to and including 59 years n=91, average age 48.1±7.4 years (from 30 years to 59 years), group II – over the age of 60 years inclusive n=146, average age 69.2±6.0 years, .For calculations, the hospitalisation rate during the year was used, which had significant differences in groups: in group I it was 0.7; in group II – 1.2 . The costs of clinical management of gout and each of the diseases that accompany it for 1 calendar year have been determined.The direct costs for the treatment of patients with gout, calculated taking into account National Clinical Protocol and comorbid pathology, increase significantly with the age of the patients: the average cost per year of the treatment of gout in patients of mature age was 1337 Euro without and 7320 Euro taking into account the concomitant pathology, and in the elderly 2067 Euro and, respectively, 15230 Euro. Cost analysis is the most important element of clinical and economic analysis, has an independent significance, including the definition of the following indicators.11The costs of treating an elderly patient with gout are largely related to the treatment of concomitant pathological conditions and complications.Direct costs are the costs incurred by the healthcare system, the patient or other payer, by society as a whole directly in the process of providing healthcare.5 Direct costs include medical and non-medical costs.Direct medical costs are direct costs for the provision of healthcare and costs associated with the provision of medical care, including those that are not directly related to the patient .6Direct non-medical costs are the expenditure of other sectors of the economy  and personal expenses of the patient and his relatives not directly related to healthcare: expenditure related to the payment for arrival at the place of treatment, with lifestyle changes due to illness.3Indirect costs are the costs of resources that were not created due to illness, the loss of society due to temporary and persistent disability and premature death.6Intangible costs are losses of patients due to suffering caused by diseases that occur but are poorly valued in monetary terms.1216With the simultaneous implementation of medical services, the question arises of the impact of the number of these services on the value of costs. In the economy, the concept of marginal cost is used, which shows additional costs when the volume of production increases by one unit of utility.9 A necessary condition for determining the costs of simultaneous healthcare in the treatment of several diseases is to take into account the fact that fixed (mainly indirect) medical costs are relatively low, and the ratio of 1/3 becomes incorrect. When developing the Nomenclature of Services in the Field of Health, the authors empirically obtained the values of the correction coefficient of the cost indicators (margin coefficient) applied in the case of calculating the costs for a complex medical service consisting of simple services performed simultaneously.14 It is proposed to introduce this coefficient in the form of a multiplier to the sum of the costs for simple medical services that are part of a complex service.The determination of average and specific  costs (costs with an increase in the number of services provided) is an important mechanism that allows you to optimize the provision of healthcare and anticipate it for a long period of time.Analysis of economic and financial efficiency in the treatment of gout in elderly patients with comorbidities.A descriptive, selective study of 237 patients with gout (average age for the men 60±8.0 years and for the women 63±9.0 years) is conducted. The study was carried out in accordance with the requirements of the Ministry of Health for “Clinical and financial-economic research” within the postdoctoral scientific program at the Discipline of Rheumatology and Nephrology of the “Nicolae Testemiţanu” State University of Medicine and Pharmacy of the Republic of Moldova.5 Determining the costs of treatment of a patient with gout in groups I and II. In order to study the clinical characteristics of gout in the elderly, all those included in the study were divided according to age at the time of examination: group I was made up of patients with gout up to and including 59 years , group II – over the age of 60 years inclusive . This age distribution is based on the recommendations of the European Consensus for patients with comorbidities. The current cost was accounted from data from a single Centre and, also, all costs are calculated from the data of compulsory medical insurance for all citizens of our country in a centralised form.From the database of the Departments of Arthrology, Rheumatology, and Nephrology of the “Timofei Mosneaga” Republican Clinical Hospital were extracted the clinic-paraclinical data and the performed treatment of patients with gout, including 658 patients with gout observed in the period 2015 – 2022, of which 237 patients who meet the criteria of the study were selected. The diagnosis of gout in the database was carried out in accordance with the classification criteria for gout according to ACR and EULAR 2015.The raw data was processed in SPSS version 26.0.Table 1). The NCP for each pathology were used separately for calculation.For the calculation we used tariffs for medical services provided under the National Clinical Protocols (NCP) which were derived from Recommendations of Guidelines of EULAR for patients with gout, taking into account comorbidities: arterial hypertension, coronary heart disease, chronic heart failure (CHF), nephrolithiasis, chronic kidney disease (CKD), diabetes mellitus type 2 (DM 2). Costs were determined for patients with gout per calendar year in the 2 study groups: in group I it was 0.7; in group II it was 1.2 (p = 0.001), including comorbid pathology , which was associated with a higher frequency of hospitalization and comorbid conditions. Inpatient treatment for chronic heart failure and ischemic heart disease was the most expensive of the comorbidities.Table 2).The most expensive of the diseases considered was the inpatient treatment of CKD and coronary artery disease.It has been calculated that treatment of gout with comorbidities costs an average of 5.5 to 29.5% of total costs per year  increases significantly compared to patients with gout of working age (0.7 cases per year), which significantly affects the cost of health care.3.The costs per case treated in the hospital for elderly gout patients, calculated in different ways, which determines the opportunity of their improvement and subsequent optimisation.


Plant viruses, as obligate intracellular parasites, induce significant changes in the cellular physiology of host cells to facilitate their multiplication. These alterations often lead to the development of symptoms that interfere with normal growth and development, causing USD 60 billion worth of losses per year, worldwide, in both agricultural and horticultural crops. However, existing literature often lacks a clear and concise presentation of the key information regarding the mechanisms underlying plant virus-induced symptoms. To address this, we conducted a comprehensive review to highlight the crucial interactions between plant viruses and host factors, discussing key genes that increase viral virulence and their roles in influencing cellular processes such as dysfunction of chloroplast proteins, hormone manipulation, reactive oxidative species accumulation, and cell cycle control, which are critical for symptom development. Moreover, we explore the alterations in host metabolism and gene expression that are associated with virus-induced symptoms. In addition, the influence of environmental factors on virus-induced symptom development is discussed. By integrating these various aspects, this review provides valuable insights into the complex mechanisms underlying virus-induced symptoms in plants, and emphasizes the urgency of addressing viral diseases to ensure sustainable agriculture and food production. Viral infections have a significant impact on plant health and can lead to a wide range of symptoms that affect growth, development, and overall productivity ,2. The eAt the molecular level, viruses have evolved sophisticated strategies to interact with and manipulate host proteins, thereby completing their life cycle within plant cells ,16,17,18In addition to directly affecting cellular processes, viral infections can trigger a series of host responses. Upon invading a plant cell, the virus utilizes the host biosynthetic machinery to synthesize its proteins and nucleic acids, while diverting the host energy and nutrient resources to locations essential to virus replication and assembly ,23,24. TThis review also discusses the influence of environmental factors on symptoms development in plants. Light and temperature are pivotal in plant physiology and are intricately linked to various signaling pathways ,31. ReceBy synthesizing these recent advances, this comprehensive review aims to provide valuable insights into the complex mechanisms underlying virus-induced symptoms in plants. Understanding these mechanisms is crucial for developing effective strategies for detecting, preventing, and controlling viral infections, ultimately safeguarding global crop production and ensuring food security in the face of emerging viral diseases.Emerging studies have revealed the mechanisms by which viruses manipulate plant cellular processes to induce these symptoms. Among the key factors involved, dysfunction of chloroplast proteins, hormone manipulation, ROS accumulation, and cell cycle control have been identified as critical contributors to the manifestation of virus-induced symptoms in plants .Emerging studies suggest a close relationship between the appearance of symptoms and alterations in chloroplast morphology and protein function. Interactions between virus-encoded proteins and chloroplast proteins play a central role in perturbing chloroplast integrity and functionality. Viruses strategically target various chloroplast components, including photosynthetic proteins, enzymes involved in chlorophyll biosynthesis, and proteins involved in carbon assimilation and metabolism ,74,75,76Nicotiana benthamiana, the excessive expression of the AltMV-encoded triple gene block 3 (TGB3) from potato virus X (PVX) leads to noticeable symptoms of veinal necrosis and chloroplast vesiculation. The interaction between AltMV-TGB3 and the Photosystem II oxygen-evolving complex protein (PsbO) is presumed to play a crucial role in symptoms development and the disruption of chloroplast structure [For example, in the case of Alternanthera mosaic virus (AltMV) infection in tructure ,81. One tructure ,83,84. CChlorotic and necrotic stripes are frequently observed in the leaves of rice plants infected with rice stripe virus (RSV), leading to subsequent wilting of the affected plants. Research findings indicate that expression of RSV-encoded disease-specific protein (SP) in transgenic rice enhanced RSV induced symptoms. This intensified effect is attributed to the interaction between SP and a component of the luminal protein complex associated with photosystem II (PsbP). In the presence of RSV SP, a substantial portion of PsbP is recruited into the cytoplasm. The absence of PsbP, as one of the key host factors involved in chloroplast function in the chloroplast, can lead to degradation of photosynthetic pigments, including chlorophyll ,86. More2O2 in the chloroplast [2O2 might be toxic to the chloroplast [Cucumber mosaic virus (CMV) infection leads to the manifestation of mosaic symptoms on various host plants. The coat protein (CP) serves as the determinant of CMV-M induced symptoms . Subsequoroplast ,88,89. Toroplast ,91. Secooroplast . As a reoroplast ,91.CHLI). Downregulation of CHLI mRNA by Y-Sat-derived siRNAs induces yellowing symptoms in infected plants [Apart from perturbing the subcellular localization of chloroplast proteins, CMV can also interfere with chloroplast genes to induce specific symptoms. The Y satellite RNA (Y-Sat) of the CMV produces short interfering RNAs (siRNAs) that target the mRNA of chlorophyll biosynthesis genes  infection on host plants are attributed to disruptions in chloroplast structure and function caused by interaction between the viral protein and host proteins. Specifically, the PVY-encoded HC-Pro interacts with the chloroplast division-related factor MinD. This interaction inhibits dimerization process of the MinD protein, consequently perturbing its functional activity and interfering with chloroplast division. This disruption leads to changes in chloroplast number within the host cells, contributing to symptoms development .These research findings provide compelling evidence that the function of chloroplast proteins plays a critical role in the development of virus-induced symptoms in plants.Hormones serve as chemical messengers, orchestrating diverse cellular responses throughout a plant lifecycle ,95,96. Bent-Kaurene oxidase leads to a reduction in the activity or transcription of ent-Kaurene oxidase. This, in turn, results in a decrease in endogenous gibberellin (GA) content within RDV-infected rice plants. The reduced accumulation of GAs directly contributes to the abnormal function of GA-regulated cellular processes, which ultimately leads to dwarf symptoms expression [Dwarf symptoms are frequently observed in rice plants infected with rice dwarf virus (RDV). The interaction between RDV-encoded P2 protein and rice pression . Additiopression .Arabidopsis thaliana ecotype Shahdara, infection with tobacco mosaic virus (TMV) induces a distinct set of disease symptoms, such as stunting, necrosis of the inoculated leaf, loss of apical dominance, and leaf curling. The 126/183 kDa replicase of TMV was found to interact with PAP1, a putative regulator of auxin response genes. This interaction disrupts the nuclear localization of PAP1, thereby affecting its transcriptional regulatory function on genes involved in the auxin response pathway. As a consequence, the gene expression patterns associated with the auxin response system are perturbed, leading to the manifestation of specific disease symptoms in infected plants [In d plants .N. benthamiana plants infected with ageratum leaf curl Sichuan virus (ALCScV) showed dwarfing symptoms and abnormal flower development. ALCScV-encoded C4 interacts with DELLA proteins (NbGAI) that act as inhibitors of plant growth and development, particularly in the GA signaling pathway, where they negatively regulate GA biosynthesis [ynthesis ,103. Theynthesis .The disruption of hormone signaling pathways by viral proteins provides insights into the mechanisms underlying the development of disease symptoms in infected plants. Understanding these interactions can contribute to the development of strategies for disease management and the improvement of crop resilience.ROS exhibits dual functionality in the invasion of pathogens in plant cells. Low levels of ROS have been demonstrated to promote signal transduction and acclimation/defense responses, whereas high levels of ROS lead to oxidation damage to lipids, DNA, and proteins ,107,108.Arabidopsis plants infected with CMV is correlated with an elevation in H2O2 production. This effect is attributed to the interaction between the CMV 2b and the isoenzyme catalase 3 (CAT3) within infected tissues, which results in the inhibition of catalase activity. This inhibition subsequently triggers oxidative stress, leading to the specific induction of necrosis [For example, the appearance of necrotic spots in necrosis .2O2 with infected tissues, consequently inducing necrosis [Maize chlorotic mottle virus (MCMV) causes chlorotic and mottle symptoms, and even necrosis when it infects maize . The MCMnecrosis .Brachypodium distachyon and N. benthamiana. Studies have revealed that BaMV infection resulted in a notable accumulation of full-length Cu/Zu superoxide dismutase preprotein (PrNbCSD2) within the cytosol, which may be achieved by impairing the cleavage of PrNbCSD2 transport peptide. This accumulation of PrNbCSD2 protein led to an excessive production of H2O2 in BaMV-infected cells, consequently resulting in the manifestation of chlorotic symptoms [Bamboo mosaic virus (BaMV) causes chlorotic mosaic symptoms in both symptoms .Mosaic symptoms caused by sugarcane mosaic virus (SCMV) infection were found to correlate with ROS accumulation in mitochondria. SCMV enhances the enzymatic activity of pyruvate orthophosphate dikinase (PPDK) at the infection front, leading to excessive production of malate under light, which in turn activates the malate circulation pathway and causes the ROS accumulation in mitochondria, thereby causing the manifestation of mosaic symptoms .Overall, these studies collectively reveal the mechanisms through which viral infections induce ROS imbalance, underscoring the significance of ROS imbalance in the pathogenic processes of viral infections.Cell division serves as the foundation for plant growth, enabling the formation of new tissues and organs throughout the plant life cycle . HoweverMembers of the plant retinoblastoma-related proteins (RBR) family serve as negative regulators of the cell cycle . SeveralIn the case of beet curly top virus (BSCTV), the viral protein C4 has been identified as the symptom determinant. C4 induces the accumulation of RKP protein, which subsequently interacts with the cell-cycle inhibitor ICK/KRP proteins. This interaction leads to alterations in the cell cycle of infected plants. However, the precise mechanism by which C4 promotes the accumulation of RKP is not yet fully understood and requires further investigation .Tomato leaf curl Yunnan virus (TLCYnV) is known to induce severe developmental abnormalities in plants. Research has shown that the C4 encoded by TLCYnV functions as one of viral symptoms determinants. This protein interacts with glycogen synthase kinase 3 (NbSKη), leading to the relocalization of NbSKη from the nucleus to the membrane and subsequently reducing its nuclear accumulation. This disruption impairs phosphorylation-dependent degradation of NbCycD1;1  ,120,121,In this section, we explored how viruses manipulate plant cellular processes to induce symptoms, focusing on dysfunction of chloroplast proteins, hormone manipulation, ROS accumulation, and cell cycle control. These alterations play crucial roles in the manifestation of virus-induced symptoms in plants, providing valuable insights into the pathogenic mechanisms of viral infections.Plant viruses induce disturbances in host metabolic processes and gene expression, manifesting as visible symptoms that hinder the growth and development of infected host plants ,125,126.One classic study investigated metabolic changes in zucchini plants infected with CMV. Spatial analysis revealed distinct metabolic alterations both at the edges of advancing lesion and inside the lesion. At the edges of the advancing lesion, there was a high demand for viral replication and synthesis of viral proteins, resulting in increased activity of anaplerotic enzymes, enhanced photosynthesis, and starch accumulation . The reaInside the lesions, there was a decrease in viral proliferation, accompanied by reduced photosynthesis and metabolism of starch. As the virus reaches its peak or starts to decline in this region, the demand for host resources decreases, leading to a reduction in photosynthetic activity . The decA study was conducted to investigate the processes underlying the symptom development in the interaction between potato plants and PVY by examining metabolite levels . In PVY-Another study focused on SCMV infection and its impact on respiratory metabolism pathways. Targeted metabolomic analysis revealed a significant induction of metabolites within respiratory metabolism pathways during the development of mosaic symptoms. Notably, there was a remarkable accumulation of malate in response to SCMV infection. Malate over-production provides energy resources for viral infection and serves as a biochemical basis for symptoms manifestation .These studies collectively demonstrate the complex interplay between viral infection and host metabolism. The metabolic changes observed in virus-infected plants are attributed to the manipulation of host metabolism by virus and the subsequent response of the host plant defense mechanisms. Simultaneously, these alterations in metabolism contribute to the development of symptoms in infected plants.The development of viral symptoms results from complex molecular and physiological processes. Thus, transcriptomic analysis of various viral infections in different plant species has provided valuable insights into the changes in gene expression associated with symptom development. Arabidopsis infected with plum pox virus (PPV), genes related to soluble sugar, starch and amino acid, intracellular membrane/membrane-bound organelles, chloroplast, and protein fate were upregulated, while genes associated with development/storage proteins, protein synthesis and translation, and cell wall-associated components were downregulated. These changes in gene expression are closely linked to the infection process and the subsequent development of symptoms [Recent studies have investigated changes in host gene expression after the appearance of viral symptoms. In symptoms . Tomato symptoms . Transcrsymptoms .Nicotiana tabacum infected with CMV revealed differential gene expression profiles. In the early stages of infection, CMV suppresses the expression of energy metabolism and pigment metabolism in the host plants. Several up-regulated pathways were related to the metabolism of terpenoids and polyketides, which might regulate plant defense [Studies have also explored changes in host gene expression at different stages of symptom development. Transcriptome analysis comparing healthy and infected plants at six stages of symptom development  in  defense . During  defense ,140. In In maize infected with SCMV, transcriptome analysis was conducted to analyze the differential gene expression in both the pre-symptomatic infection stage and the steady-symptom infection stage. In pre-symptomatic infection, the function of DEGs is mainly related to protein processing in the endoplasmic reticulum, for fatty acid biosynthesis, and for RNA splicing. During steady-symptom infection stage, the function of DEGs is mainly related to proteins of the thylakoid and other plastid-related cellular components . ThroughBased on the similarity of transcriptional changes in different viruses during the induction of host symptoms development, it is suggested that the increased energy demand during viral infection has a significant impact on energy metabolism, respiration, and photosynthetic rates. The increased energy demand during viral infection has a significant impact on energy metabolism, respiration, and photosynthetic rates. This leads to the accumulation of ROS, which can directly induce viral symptoms and act as stress signals to initiate defense pathways. Moreover, the disruption of phytohormonal balances due to shared intermediates between stress signaling and hormone synthesis greatly affects the physiology of the host, possibly related to photosynthetic rates and chloroplast reactions. The cumulative effects of inhibited chloroplast functions may be associated with stunted growth and poor development of leaves .The host metabolic and genetic responses to virus-induced symptoms are complex processes that involve alterations in metabolic pathways and changes in gene expression. Viral infections strategically manipulate host metabolism to support their replication, leading to changes in carbohydrate utilization, ROS accumulation, and energy production. These alterations, along with shifts in gene expression, contribute to the manifestation of visible symptoms in infected plants. Understanding these host responses provides valuable insights into the pathogenesis of viral infections and can aid in the development of effective disease management strategies.Light and temperature are important environmental factors that profoundly influence plant growth, development and physiology. Interestingly, many studies have described the influence of light and temperature on development of viral symptoms. This section summarizes the effects of light and temperature conditions on the development of viral symptoms .Studies investigating the effect of light conditions on viral symptoms development have revealed intriguing findings. In the case of potato mop-top virus (PMTV) infection in tobacco plants, it was observed that the appearance of necrotic spots or small rings was delayed when there was a decrease in light intensity or photoperiod. When inoculated plants are transferred from light to darkness, necrotic rings develop, and the rate of virus accumulation increases. However, when the order of the treatments is reversed, no lesions appear. Therefore, the process of PMTV-induced lesion formation includes an early phase requiring light .Similarly, a field survey conducted in an unmanaged forest revealed interesting observations regarding viral symptoms in shade-grown and sun-exposed plants. Virus-infected plant species growing under shade exhibited fewer apparent viral symptoms, while those growing in direct sunlight displayed severe chlorosis in their leaves .To understand these observations, it is important to consider the role of light in photosynthesis. When a plant is infected with a virus, it can affect the photosynthetic machinery, leading to changes in energy production ,74. AlteThe above research indicates the intricate relationship between light conditions and the development of symptoms. It is worth noting that the specific effects of light on plant virus symptoms can vary depending on the virus-host combination, light intensity, duration, and other environmental factors. The specific mechanism by which light affects the development of virus symptoms is still being investigated, but these findings highlight the importance of considering light conditions when studying plant-virus interactions and symptoms development.Temperature is a crucial factor in determining the outcome of plant virus infections. However, the effects of increased temperatures on viral symptoms can vary depending on the specific virus-host combinations. Some studies have shown that higher temperatures can intensify virus symptoms , while iN. tabacum plants infected with CMV, higher incubation temperatures (28 °C compared to 18 °C) resulted in more severe symptoms. Molecular analysis of CMV-infected plants revealed that at lower temperatures, several genes associated with salicylic acid (SA) were upregulated, while at higher temperatures, genes associated with jasmonic acid (JA) showed increased expression [For example, in pression . SA-depeN. benthamiana plants. However, when the temperature drops to 15 °C, these symptoms progress to systemic necrosis, resulting in plant death within 30 dpi. By contrast, a chimera consisting of the CP derived from a mild AltMV strain only induces systemic mosaic symptoms at 15 °C, reducing the severity of the symptoms caused by the severe strain. Surprisingly, there was no significant difference in virus accumulation between the severe and mild strains at a given temperature. However, it was observed that both strains exhibited significantly higher virus accumulation at 15 °C when compared to 25 °C [At 25 °C, a severe strain of AltMV causes mosaic patterns and localized necrosis in to 25 °C . This ob2 concentration, UV radiation, ozone, and drought, can also influence the interactions between plant and virus [These studies indicate that light and temperature can have diverse effects on plant viral infections and the severity of associated symptoms. The interplay between environmental factors, plant defense pathways, and viral factors contributes to the complex molecular mechanisms underlying these phenomena. Nevertheless, it is essential to note that beyond light and temperature, other environmental factors, such as COnd virus ,152,153.Understanding the complex mechanisms by which viruses interact with host factors and manipulate cellular processes is essential for unraveling the causes of virus-induced symptoms in plants. Recent research has provided valuable insights into these intricate processes, revealing the mechanisms by which viral infections disrupt chloroplast protein function, hormone regulation, ROS accumulation, and cell cycle control. Additionally, host reactions such as metabolic disorders and gene expression changes contribute to symptoms development. The influence of environmental factors on viral infection and symptoms severity further emphasizes the multifaceted nature of virus-plant interactions. By integrating these findings, this comprehensive review offers a holistic understanding of the mechanisms underlying virus-induced symptoms in plants, paving the way for the development of effective strategies for disease management and crop protection strategies.By gaining a deeper understanding of how viruses disrupt plant cellular processes, novel intervention strategies can be explored to mitigate the impact of viral infections on crops. Building upon our understanding of the metabolic disorders and gene expression changes induced by viral infections, targeted approaches can be developed to enhance the crops resistance to diseases. One of the cutting-edge technologies that has emerged in recent years is the use of RNA-based approaches, such as dsRNA and siRNA gene interference, microRNA, and CRISPR/Cas9. These RNA-based techniques offer promising methods for developing virus-resistant crop plants. By introducing specific RNA molecules that target viral genes or disrupt viral replication, these approaches can effectively inhibit viral infections in plants. Moreover, the development of non-transgenic methods, such as topical application of dsRNA, hairpin RNA, and artificial microRNA, provides GMO-free virus disease management options, which are increasingly sought after by consumers and regulatory authorities ,155. AnoIn conclusion, through in-depth research on virus-plant interactions, we will gain a better understanding of the mechanisms underlying virus-induced symptoms in plants and provide more effective strategies for disease management and crop protection. This will have significant implications for the sustainable development of agriculture and food security.


Benincasa hispida var. chieh-qua How) is an important crop in South China and southeast Asian countries. Viral diseases cause substantial loss of chieh-qua yield. To identify the viruses that affect chieh-qua in China, ribosomal RNA-depleted total RNA sequencing was performed using chieh-qua leaf samples with typical viral symptoms. The virome of chieh-qua comprises four known viruses (melon yellow spot virus (MYSV), cucurbit chlorotic yellows virus (CCYV), papaya ringspot virus (PRSV) and watermelon silver mottle virus (WSMoV) and two novel viruses: cucurbit chlorotic virus (CuCV) in the genus Crinivirus and chieh-qua endornavirus (CqEV) in the genus Alphaendornavirus. The complete genomes of the two novel viruses in chieh-qua and three other isolates of CuCV in pumpkin, watermelon and cucumber were determined and the recombination signals of pumpkin and watermelon isolates of CuCV were detected. A reverse transcriptase PCR indicated that the dominant viruses of chieh-qua in Hainan are MYSV (66.67%) and CCYV (55.56%), followed by CuCV (27.41%), WSMoV (7.41%), cucumber mosaic virus (8.15%), zucchini yellow mosaic virus (6.67%), PRSV (6.67%) and CqEV (35.56%). Our findings support diagnostic and prevalence studies of viruses infecting chieh-qua in China, enabling sustainable control strategies for cucurbit viruses worldwide.The cucurbit vegetable chieh-qua ( Benincasa hispida Cogn. var. chieh-qua How), also known as fuzzy gourd, hairy melon or moa qua, is a variety of wax gourd (Benincasa hispida (Thunb.) Cogn.) in the Cucurbitaceae family. As an important vegetable crop, chieh-qua is widely cultivated throughout the world for its immature fruit, especially in South China and southeast Asian countries [Benincasa hispida was about 6 million acres, with a yield of 58 million tons [Chieh-qua , zucchini yellow mosaic virus , papaya ringspot virus  and cucumber mosaic virus  [However, with the expansion of cultivation, various diseases have arisen, especially viral diseases, which have been a major factor restricting the yield and quality of chieh-qua in China. Diverse virus-like symptoms, including mosaic, interveinal chlorosis, mottling and leaf deformation, are frequently observed in chieh-qua; however, there has been limited research on sporadic chieh-qua samples and only four viruses have been reported: zucchini tigre mosaic virus  ,7,8. ZYMmovirus) , whereasmovirus) ,10. Recemovirus)  investigCrinivirus and Endornavirus, whose complete genomes and natural hosts within gourd vegetables were also determined. Furthermore, an extensive virus screen was conducted on over two hundred samples collected from various plantations in Hainan province from 2021 to 2022. This study provided comprehensive information on the virus diversity of chieh-qua in China, the prevalence and geographical spread of viruses infecting chieh-qua and the dominant viruses in different plantations.The present study aimed to investigate in depth the viruses causing chieh-qua viral disease in China. For this, a ribosomal RNA (rRNA)-depleted total RNA method was used to construct the virome profiles of symptomatic chieh-qua samples. This approach identified four known viruses , PRSV and WSMoV) and two novel viruses in the genus Cucurbita moschata) samples, 26 watermelon (Citrullus lanatus) samples and 49 cucumber (Cucumis sativus) samples, were collected to determine whether the potential new viruses occur in other cucurbit vegetables. The above 223 symptomatic samples and 28 asymptomatic chieh-qua samples were collected from 7 main cucurbit vegetable cultivation regions  in Hainan province from 2021 to 2022  and its integrity was evaluated using a 2100 Bioanalyzer . An Epicentre Ribo-Zero Gold Kit  was used to deplete ribosomal RNAs from the total RNA samples. An Illumina TruSeq® RNA Sample Prep Kit  was then used to construct a cDNA library from the ribosomal RNA-depleted RNA. The cDNA library was then subjected to HTS using the PE150 sequencing platform on an Illumina NovaSeq 6000 sequencer , carried out by LC-Bio Technology Co., Ltd. .Leaves from five individual chieh-qua plants  representing the distinct symptoms B were poBenincasa hispida Cogn. var. chieh-qua How has not been published; therefore, the genome sequence of Wax gourd (Benincasa hispida)  was used as the reference to eliminate host-derived sequences using HISAT v2.1.0 [−4 and 10−6, respectively.The raw reads from rRNA-depleted total RNA sequencing were trimmed to remove low-quality and adapter sequences using fastp . The genT v2.1.0  was usedT v2.1.0  searchesT v2.1.0 , employiEscherichia coli DH5α competent cells . We then selected three individual clones of each amplicon that had inserts of the expected size, which were sequenced in both directions using Sanger sequencing, carried out at BGI .An RNAPrep Pure Plant Kit  was used to extract the total RNA from individual plants, which was reverse transcribed into cDNA using random hexamers and TransScript One-Step gDNA Removal and cDNA Synthesis SuperMix  following the supplier’s recommendations. The 2×Rapid Master Mix  was used to carry out subsequent PCR reactions using specific virus detection primers . These pClosteroviridae was obtained with complete 5′ and 3′ untranslated regions (UTRs). A SMARTer® RACE 5′/3′ kit with SeqAmp DNA Polymerase  was used to determine the 5′ UTR sequence of the novel crinivirus. Poly(A) Polymerase  was then used to add additional ATPs to the 3′ terminus of the viral genome lacking a poly(A) tail following the supplier’s guidelines. Subsequently, the 3′ terminal sequence was completed using an anchored oligo (dT) primer (5′-TGTGTTGGGTGTGTTTGGTTTTTTTTTTTTTTT-3′) to synthesize the cDNA from the poly(A)-tailed RNA virus, followed by amplification using the primer anchored-dt-rev (5′-TGTGTTGGGTGTGTTTGG-3′) together with a gene-specific primer.Following confirmation of their virus status, the whole genomes of the newly discovered viruses were amplified using 2× Phanta Flash Master Mix  with various pairs of virus-specific primers . The seqhttps://www.ncbi.nlm.nih.gov/orffinder/, accessed on 2 December 2022) was used to predict the open reading frames (ORFs) and the Conserved Domain Database  was used to analyze conserved motifs and domains. Transmembrane domains were predicted using DeepTMHMM [For genomic characterization, the web tool ORFfinder  isolates  and the CYSDV isolates available in the NCBI database using the recombination analysis software SimPlot version 3.5.1  and the De novo assembly of the remaining 4,018,906 (4.93%) unmapped reads produced 23,111 contigs . BLASTn and BLASTx analyses identified 31 contigs corresponding to viruses of the Closteroviridae family , Tospoviridae , Potyviridae  and Endornaviridae  , whose Q20 and Q30 values were >94%. A total of 77,465,294 (95.07%) of the reads mapped to the wax gourd reference genome. ,020 nt) . AccordiTo confirm that the putative viruses were not artefacts of HTS sequencing, seven primer pairs  were desThe entire genomes of the new crinivirus, CuCV, in chieh-qua were 9157 nt for RNA1 and 7788 nt for RNA2, whose poly(A) tails were of an undefined length A. To betBLASTn analyses revealed that CuCV and a so-called CYSDV Thailand isolate (MT813029 and MT819949) available in the GenBank database are molecularly identified as the same virus species. This conclusion is based on their complete genomes sharing more than 98% nucleotide identity (with 99% coverage). However, they exhibit significant divergence from other CYSDV isolates, with nucleotide identities ranging from 70.25% to 74.05%  and a shared amino acid identity of 74.1% for the coat protein . These fCrinivirus [Crinivirus, a putative P-Pro domain [The genomic organization of the characterized virus A was verinivirus . The RNAo domain  , a heat-shock-protein-70-like protein , p6 , p59 , p9 , coat protein , minor CP  and p26 . Neither the p5 in RNA1 nor p4 in RNA2 have any homologs in the database. The seven nucleotides (ACATGGG) at the 5′-UTR of RNAs 1 and 2 are identical and although this feature is common in criniviruses , this seCrinivirus in the Closteroviridae family revealed that four CuCV isolates  in this study and the so-called CYSDV isolate from Thailand were consistently placed in a branch distinct from other CYSDV members  and the formally identified criniviruses showed that the CuCV isolates were located in a distinct cluster away from the CYSDV clusters, which, similar to the results of previous studies , was sep−32 according to the 3Seq method and a high recombination score of 0.815, which targeted the region encoding the replication-associated protein. The break points of the watermelon-LS isolate started at nt 2170 and terminated at nt 8686, with the highest probability value of 10−12 according to the SiScan method and a recombination score of 0.526  has a monopartite genome of ~15,020 nt based on the long scaffolds that were assembled from the rRNA-depleted total RNA sequencing, which lacks 5′- and 3′-terminal sequences (GenBank accession number OQ851472). Similar to other endornaviruses, the CqEV genome has only a single ORF encoding a 4981 aa polyprotein  A. A consAlphaendornavirus, phylogenetic analysis of RdRP protein sequences from different endornaviruses (Alphaendornavirus and formed a sister branch to CmEV (KT727022). Furthermore, the genome sequence of CqEV was compared with those of the 24 reported formal species in the genus Alphaendornavirus. The results indicated that CqEV is most closely related to CmEV (KT727022), sharing a 65.41% nt identity  and the new endogenousvirus (CqEV) in different cucurbit vegetables, 98 symptomatic samples of 4 cucurbit vegetables were collected from 7 counties of Hainan. A total of 16 samples were CuCV positive by RT-PCR, including samples from pumpkin (4/23), watermelon (4/26) and cucumber (8/49) . No CqEVBemisia-tabaci-transmitted crinivirus causing extensive infections of cucurbit crops in many warm and temperate production areas [Closteroviridae.In this study, a putative novel crinivirus was identified and molecularly characterized. The closest relative of CuCV appears to be CYSDV, a on areas ,31,32,33Closteroviridae [Five CuCV isolates (four from this study and one from Thailand) from different natural hosts  showed generally limited spatial and temporal sequence variability, which is also found in some other viruses in the family oviridae ,34,35,36In this study, a strong association between CuCV and specific symptoms was suggested because CuCV was found exclusively in symptomatic cucurbit samples but not in any of the 28 asymptomatic samples, which aligns with the results reported by Krishnan et al.  where CuCuCV is the fourth crinivirus reported to infect cucurbit plants in addition to CYSDV, CCYV and beet pseudoyellows virus (BPYV) ,36. So fVicia faba endornavirus [CqEV is the first endornavirus reported to infect chieh-qua. No CqEV was found as a single infection in symptomatic samples and 9 out of 28 asymptomatic samples were positive for CqEV, which indicated that CqEV is in a symbiotic relationship with chieh-qua causing no severe symptoms, similar to most plant endornaviruses, with the exception of rnavirus ,40,41. Crnavirus , the majrnavirus . Althougrnavirus , the natIn this study, chieh-qua is for the first time reported as the natural host of CuCV, MYSV and CqEV. Complex mix infection was commonly found in chieh-qua, though 57 samples were “single infected”. The virus-induced symptoms cannot differentiate between single or multiple infections, since the single-infected samples might have been infected with other unknown viruses besides the tested ones. Among the identified viruses, MYSV and CCYV were the most prevalent in this study and they were also reported to cause great losses in cucurbit vegetable production in many countries ,47,48,49


Agaricus subrufescens is considered as one of the most important culinary-medicinal mushrooms around the world. It has been widely suggested to be used for the development of functional food ingredients to promote human health ascribed to the various properties . In this context, the interest in A. subrufescens based feed ingredients as alternatives for antibiotics has also been fuelled during an era of reduced/banned antibiotics use. This study aimed to investigate the effects of a fermented feed additive -rye overgrown with mycelium (ROM) of A. subrufescens—on pig intestinal microbiota, mucosal gene expression and local and systemic immunity during early life. Piglets received ROM or a tap water placebo (Ctrl) perorally every other day from day 2 after birth until 2 weeks post-weaning. Eight animals per treatment were euthanized and dissected on days 27, 44 and 70.Undibacterium and Solobacterium) and caecum (Intestinibacter and Succinivibrionaceae_UCG_001) on day 70, as compared to Ctrl piglets. ROM supplementation also influenced gut mucosal gene expression in both ileum and caecum on day 44. In ileum, ROM pigs showed increased expression of TJP1/ZO1 but decreased expression of CLDN3, CLDN5 and MUC2 than Ctrl pigs. Genes involved in TLR signalling  were more expressed but MYD88 and TOLLIP were less expressed in ROM pigs than Ctrl animals. NOS2 and HIF1A involved in redox signalling were either decreased or increased in ROM pigs, respectively. In caecum, differentially expressed genes between two groups were mainly shown as increased expression  in ROM pigs. Moreover, ROM animals showed higher NK cell activation in blood and enhanced IL-10 production in ex vivo stimulated MLN cells before weaning.The results showed ROM piglets had a lower inter-individual variation of faecal microbiota composition before weaning and a lower relative abundance of proteobacterial genera in jejunum  immune system development. Consequently, ROM supplementation may contribute to improving health of pigs during the weaning transition period and reducing antibiotics use.The online version contains supplementary material available at 10.1186/s42523-023-00244-w. Pigs are exposed to various stressors during their early life in modern commercial farming, with weaning as one of the most stressful events in a pig’s life. The weaning associated psychological, environmental and nutritional stressors have the potential to disrupt the pig’s gastrointestinal (GI) health, increase disease susceptibility, thereby compromising animal health, welfare and growth performance . RecentlAgaricus subrufescens (A. subrufescens) through the process of solid-state fermentation. During the fermentation procedure, A. subrufescens produces enzymes  that assist in the degradation and utilization of substrate (rye) nutrients  into indigestible but bioactive oligosaccharides _coprostanoligenes_group, but lower relative abundance of Peptostreptococcus than Ctrl animals before weaning , as well as jejunal, ileal and caecal luminal digesta at each dissection day . As expected, time  together with the diet switch from milk to solid feed were the two major drivers for faecal microbiota development, both with respect to alpha- as well as beta- diversity  and pH (p = 0.04) also significantly contributed to explaining the observed variation in ileal gene expression, and significant interactions were found between BW × time (p = 0.02) and pH × time (p = 0.001). When we analysed ileal gene expression data separately for the different sampling days, the variables treatment (p = 0.05), ileal pH (p = 0.001) and BW (p = 0.02) all contributed to explaining the observed variation in ileal mucosal gene expression on day 27  and ileal pH (p = 0.03) significantly contributed to explaining the variation of ileal mucosal gene expression on day 44   and day 44 (16 days post-weaning). Of the 96 selected genes, 78 were considered expressed in the ileum and 79 in the caecum. Redundancy analysis (RDA) revealed a strong effect of sampling day in both tissues  as well as serum amyloid A genes , all of which are known to be induced by inflammatory stimuli, was found to be negatively associated with ROM treatment Fig. A, B. In Analysis of differentially expressed genes (DEG) on day 44 versus day 27, showed several ileal and caecal mucosal genes were differentially expressed post-weaning in both Ctrl and ROM-treated groups compared to pre-weaning Fig. A–D. A laIn ileal mucosa of Ctrl pigs three of the five DEGs with increased expression on day 44 were MUC2 and TFF2 and TFF3, which are expressed in goblet cells and involved in epithelial barrier function and restitution. Additionally, CCL5 and TLR3 were upregulated in the ileum of the Ctrl group after weaning on day 44. In the ileum of ROM treated pigs several genes involved in TLR signalling , inflammatory responses , redox signalling (HIF1A and GSTM3) and apoptosis pathways  showed higher expression levels on day 44 as compared to day 27. In addition, the membrane bound mucin encoding gene MUC1 and the short-chain fatty acid transporter encoding gene SLC5A8 were upregulated in the ileum of ROM treated pigs post weaning.2, PDGFRB) as well as TNF. DEGs which were upregulated in the caecum of Ctrl pigs when comparing day 44 and day 27 included genes involved in TLR signalling , encoding cytokines and chemokines  as well as BAX and RETNLB. In the caecum of ROM treated pigs, genes encoding membrane associated mucin MUC1, LPS binding protein (LBP) and BAX were down-regulated between day 44 and 27. Two genes related to TLR signalling , cytokines  and genes involved in apoptosis pathways  were upregulated after weaning in the cecum of the ROM treated group.In the caecal mucosa of Ctrl pigs, weaning (day 44 vs. day 27) led to a decrease in expression of MUC2, the glucose transporter encoding gene GLUT3, genes involved in TLR signalling MyD88, genes negatively regulating inflammatory signalling , genes encoding the epithelial tight junction claudins (CLDN3 and CLDN5), genes related to redox balance  were more highly expressed in the caecum of the ROM group than the Ctrl group, whereas the expression levels of BAX and MUC1 were lower in caecal mucosa of ROM animals than in Ctrl pigs.Streptococcus was negative correlated with the gene NFKB1 on day 27 , TICAM2 and IRAK4, respectively  and TFF3, respectively  vaccination  of Salmonella-specific IgM when including all time points. Furthermore, MLN cells from ROM-treated animals produced higher levels of IL-10 upon stimulation with 10 µg/mL LPS  in comparison to LPS stimulated MLN cells from Ctrl animals . No such differences were observed for the post-weaning time points . Moreover, NK cells from ROM-treated animals were more activated than NK cells from Ctrl animals, considering all time points (p = 0.005). As for single time point analysis, a significant difference between the treatment groups was only observed on day 26 pre-weaning (p = 0.017)  Fig. C.Fig. 8IUsing feed additives as alternatives for antibiotics to shape gut functioning and host immune competence has received an increasing interest for piglets in suckling and nursery phases , 44. HowEubacterium]_coprostanoligenes_group, whereas Peptostreptococcus was significantly more abundant in Ctrl piglets during pre-weaning. The abundance of [Eubacterium]_coprostanoligenes_group has previously been reported to positively respond to dietary Bacillus subtilis supplementation for piglets around weaning [Peptostreptococcus may include potential porcine intestinal pathogens and was associated with the level of Clostridioides difficile in suckling piglets [Proteobacteria, including Undibacterium and Solobacterium, were only present in Ctrl pigs in the jejunum on day 70, and two other proteobacterial genera (Intestinibacter and Succinivibrionaceae_UCG_001) were less abundant in ROM pigs in the caecum on day 70. This reduction in members of Proteobacteria in ROM pigs in jejunum and caecum on day 70 may be considered beneficial as the increased prevalence of this phylum is a potential signature of intestinal dysbiosis [Methanosphaera was only present in ROM pigs in the caecum on day 70. A higher abundance of Methanosphaera spp. in pigs has been reported to be associated with fat metabolism [As hypothesised, ROM supplementation affected the development of piglet gut microbiota, but the modulation was modest. Pigs supplemented with ROM tended to have a lower alpha diversity of faecal microbiota compared to Ctrl piglets during pre-weaning, as based on observed richness and InvSimpson. These alpha diversity indices emphasise on taxa richness and evenness , 48, res weaning , and be  weaning . In cont piglets . Intriguysbiosis . Converstabolism , howeverThe intestinal mucus layer is one of the first lines of intestinal protection. Mucins expressed by epithelial cells are the major structural and functional components of mucus –59. In tTight junctions (TJs) play a critical role in regulating the permeability of the intestinal barrier . WeaningThe epithelial TLR/IL-1R signalling pathway has been reported to play a crucial role in regulating gene expression and shaping host immune response in the intestine, and thus helping maintaining intestinal homeostasis , 76. WeaGenes related to redox signalling and cell apoptosis were also modulated by ROM supplementation. Weaning stress has been reported to cause increased oxidative processes and then lead to enterocyte apoptosis, which negatively affects intestinal barrier function , 44. In Salmonella-specific IgM was observed, which was already visible prior to vaccination (day 14). This may imply that Salmonella-specific antibodies were already present in the pigs’ blood or that antibodies to other Enterobacteriaceae cross-react with components of the ELISA. It has been suggested that the effects on the innate immune system are induced by pathogen-associated molecular patterns (PAMPs) such as β-glucans. PAMPs like β-glucans can interact with pattern recognition receptors (PRRs) on the surface of intestinal epithelial cells and innate immune cells , thereby eliciting an immune response through specific signalling pathways [A. subrufescens augmented NK cell cytotoxicity through IL-12-mediated IFN-γ production [It is known that feed additives may influence the immune system either directly, or indirectly through the gut microbiota and epithelial barrier , 94. In pathways , 96. Thipathways . Anotheroduction . Though,A. subrufescens mycelia and/or rye during solid state fermentation. Recent studies have specified that antitumor, anti-inflammatory, antioxidant and antiallergic effects of the mycelium extract (Andosan™) mainly made from A. subrufescens are presumably due to β-glucans and low molecular weight (LMW) substances , although the quantity of the β-glucans is small and the main constituent of Andosan™ is still not fully resolved [A. subrufescens was also observed in an in vitro assay [A. subrufescens could also induce apoptosis as a mechanism to inhibit tumour cell growth both in vitro and in vivo [A. subrufescens mycelium. However, some of the potentially beneficial effects from ROM could also be derived from rye compounds. Arabinoxylans as the main non-digestible fibre of cereal grains (rye included), were found to modulate gut microbiota and their metabolites  as well as to enhance immune responses [The observed effects of ROM supplementation early in life on piglet intestinal microbiota composition, epithelial gene expression, and host immune response could be linked to functional components either derived from resolved , 98. A presolved –101. An ro assay , and in ro assay , 103. Ad in vivo , 105. Inesponses , 107.Taken together, the results observed in this study provide evidence that ROM supplementation for pigs in early life can positively modulate gut microbiota, intestinal mucosal function, and immune system development. This may contribute to improving health of pigs during the weaning period and further reducing antibiotics use. Further studies are required to clarify the constituents and functional compounds present in ROM, and to confirm if ROM supplementation in early life can indeed increase resilience of piglets against  infections during the weaning period.Additional file 1: Table S1. Weaner and nursery diet. Table S2. Primers used for microfluidic qPCR. Fig. S1. The effect of ROM oral supplementation for pig on body weight in over time (A), average daily feed intake (B) and feed conversion ratio (C) in nursery phase. Control (Ctrl) and treatment (ROM) groups are represented by colour blue and yellow, respectively. Data is shown as the means ± the standard error of the mean (SEM). No significant differences on either body weight, average daily feed intake or feed conversion ratio were observed between both groups. Fig. S2. Comparison of faecal microbial alpha diversity between ROM and Ctrl groups over time, with metrics of observed richness (A), phylogenetic diversity (B), Shannon diversity (C) and inverse Simpson (InvSimpson) (D). Data was separated according to weaning for Linear Mixed-Effects Model analysis and no statistically significant differences were observed between the two treatment groups either during pre- or post-weaning phase, with any index of alpha diversity. Treatment (ROM) and control (Ctrl) groups are represented by colour yellow and blue, respectively. The vertical line indicates weaning on day 28. Fig. S3. Principal coordinate analysis (PCoA) of faecal microbial composition over time, based on unweighted UniFrac (A) and weighted UniFrac (B) distance metrics. Data was separated according to weaning on day 28 to assess the effect of time and treatment on microbial variation using PERMANOVA. Colours represent different time points, and shapes represent different treatment groups . The percentages at the axes indicate the variation explained. Fig. S4. Genera showing tendencies to be differential abundant between ROM and Ctrl pigs during pre-or post weaning. Genera were identified through GAMLSS-BEZI model with random effect. The p value was corrected by FDR for multiple testing. The yellow and blue colours represent treated (ROM) and control (Ctrl) groups, respectively. Fig. S5. Comparisons of gut luminal microbial alpha diversity between ROM) and Ctrl groups over time. The jejunal, ileal and caecal luminal microbial alpha diversity are shown in (A–D), (E–H) and (I–L), respectively, with metrics of observed richness, phylogenetic diversity, Shannon diversity and inverse Simpson (InvSimpson), from left to right. Differences were assessed with a Linear Mixed-Effects Model for jejunal, ileal and caecal luminal microbial alpha diversity for all time points, respectively. Treatment (ROM) and control (Ctrl) groups are represented by colour yellow and blue, respectively. Fig. S6. Principal coordinate analysis (PCoA) plots for the overall microbial composition of intestinal luminal digesta at different locations, based on unweighted- and weighted UniFrac metrics. Significance of the effect of time and treatment on jejunal- , ileal-  and caecal  luminal microbiota at ASV level was assessed by PERMANOVA. Colours represent different time points, and shapes represent different treatment groups . The percentages at the axes indicate the variation explained. Fig. S7. Bar plots representing relative abundance in individual pig jejunal (A–C) and caecal (D–F) luminal contents of specific taxa identified as differential features by LEfSe for pigs between control (class: Ctrl) and treatment (class: ROM) on day 70. The solid black horizontal line indicates the mean relative abundance within each class. Fig. S8. Redundancy analysis (RDA) tri-plots for the association between the variation of gene expression and environmental variables of time, treatment, pH and BW. A and B illustrates the association between ileal and caecal mucosa expressed genes with environmental variables of time, treatment, pH value and body weight (BW), respectively. C shows the association between the caecal mucosal gene expression and treatment, pH values and BW. Blue and yellow colours represent control (Ctrl) and treated (ROM) groups and shapes show the different time points. Dark grey arrows indicate environmental variables and light grey arrows show best fitting genes in the model. The percentages at the axes indicate the variation explained by the first two canonical axes. Fig. S9. Correlation analysis between the relative abundance of predominant ileal luminal genera and mucosal gene expression on day 27. The orange and blue coloured circles correspond to a positive and negative statistical correlation with an adjusted p value < 0.05, respectively. Insignificant correlations were shown as blank. Fig. S10. Correlation analysis between the relative abundance of predominant ileal luminal genera and mucosal gene expression on day 44. The orange and blue coloured circles correspond to a positive and negative statistical correlation with an adjusted p value < 0.05, respectively. Fig. S11. Correlation analysis between the relative abundance of predominant caecal luminal genera and mucosal gene expression on day 27. The orange and blue coloured circles correspond to a positive and negative statistical correlation with an adjusted p value < 0.05, respectively. Insignificant correlations were shown as blank. Fig. S12. Correlation analysis between the relative abundance of predominant caecal luminal genera and mucosal gene expressions on day 44. The orange and blue coloured circles correspond to a positive and negative statistical correlation with an adjusted p value < 0.05, respectively. Insignificant correlations were shown as blank.


For many wound carers globally getting a handle on the costs associated with the management of chronic wounds is difficult, if not impossible. There are some published figures geographically but most of these are not ‘standardised’ to permit comparison easily or directly. One consistent theme from a number of these studies, however,While most of these studies capture or measure their wound care costs differently making a direct comparison between geographies difficult, they at least provide some measure to permit a comparison. Another weakness of any comparison between studies or geographies is the time frame of measurement. Indeed, a simple literature search shows both the paucity of data generally, in some geographies and the outdatedness of the data in others. Cost studies are tedious, difficult, lengthy and expensive, as such they are seldom carried out.IWJ editors decided to provide an annual appraisal of the likely wound care costs by geographical area around the world.Due to the difficulties of capturing such cost data most of these studies caution their results as an underestimate of the costs involved.The data presented in Table EWCE—estimated wound care expenditure —our estimate of the likely wound care costs.PCHCS—per capita health care spend current published per capita healthcare cost. For the purposes of this editorial, we focused on 2019 to eliminate the bias related to COVID‐19 costs. We will jump ahead to 2022 when they are published.TP is total population.AWCCP—average wound care cost percentageseveral published studies have indicated that the percentage of total healthcare costs that is represented by the cost of wounds, ranges from 2% on the low end to 5% on the high end. For the purposes of our calculations, remembering different geographies can be at differing evolutionary stages regarding wound care, we chose the median of 3.5% as the AWCCP.Here are how the figures were calculated:While our computation may not be highly scientific, it provides an indication of the likely costs of wounds. These figures can be validated by others locally, but we feel they provide a starting point for those requiring some indication of their costs.We welcome any input from those involved in such costing studies around the world, perhaps providing a deeper insight. We are happy for others to refine our figures geography by geography providing a more accurate costing where possible.The authors declare no conflict of interest.


Studies suggest that chronic hepatitis C patients who achieve sustained virologic response (SVR) have lower risks of liver-related morbidity and mortality. Given the substantial costs and complexity of hepatitis C virus (HCV) antiviral treatment, post-treatment benefits are important to understand.To determine whether health care costs and utilization for up to 5 years after treatment differed between patients who achieved SVR and those who did not.Kaiser Permanente Medical Care Program patients receiving HCV treatment with pegylated interferon and ribavirin (Peg-IFN/RBV) from 2002 to 2007 were retrospectively analyzed, excluding those with human immunodeficiency virus (HIV) or chronic hepatitis B. Health care utilization and costs for up to 5 years after treatment completion were derived from electronic records. We compared mean annual cost and overall post-treatment costs (standardized to year-2007 dollars), and yearly utilization counts between the SVR and non-SVR groups, adjusting for pretreatment costs, age, sex, baseline cirrhosis, and race using gamma and Poisson regression models.The 1,924 patients eligible for inclusion were a mean age of 50 years; 63% male; 58% white, non-Hispanic; 62% with genotype 1; and 48% who had achieved SVR. The mean duration of post-treatment time was 3 years, and patients without SVR incurred significantly higher health care costs than patients with SVR. For each post-treatment year, total adjusted costs were significantly higher in the non-SVR group than in the SVR group, with rate ratios (RRs) and 95% CIs ranging from 1.26  to 1.64 , driven mostly by hospital and outpatient pharmacy costs. When all post-treatment years were considered collectively, the non-SVR group had significantly higher costs overall  and in each category of costs. The adjusted difference in yearly total mean costs was $2,648 . In post-treatment years 2-5, adjusted liver-specific laboratory test rates were 1.8 to 2.3 times higher in the non-SVR group than in the SVR group . During post-treatment years 1-5, adjusted yearly liver-related hospitalization rates were up to 2.45 times higher , and medicine/GI clinic visit rates were up to 1.39 times higher  in the non-SVR group compared with the SVR group.Health care utilization and costs after HCV antiviral therapy with Peg-IFN/RBV, particularly for liver-related tests, outpatient drugs, and hospitalizations, were significantly lower for patients who achieved SVR than for those without SVR. Our observations are consistent with the potentially lower risk of severe liver disease among patients with SVR.


We present a case of a 57-year-old man with newly diagnosed acquired immunodeficiency syndrome (AIDS) infection who initially sought care for progressive dysarthria and gait instability. Neuroimaging and CSF studies revealed a diagnosis of progressive multifocal leukoencephalopathy (PML). Although the patient’s human immunodeficiency virus (HIV) decreased considerably in response to anti-retroviral therapy, he continued to deteriorate clinically. Ultimately, the central nervous system (CNS) lesions, which were once centered in the cerebellum, became expansile throughout his posterior fossa. There are few reported cases of cerebellar PML in patients with AIDS. PML is a CNS demyelinating disease caused by the John Cunningham (JC) virus, which is a double-stranded DNA virus also known as Polyomavirus 2. JC virus infection is usually lifelong and asymptomatic. However, it may become pathogenic in immunodeficient patients. The IgG seroprevalence for JC virus has been calculated around 58% and it may increase with age The patient presented to an outside hospital in March of 2022 with gait disturbances. He was found to have HIV, with a viral RNA of 315,000 copies/mL, and an MRI that revealed T2 hyperintense lesions within the left cerebellum and peduncle. He declined antiretroviral therapy, left against medical advice, and presented to our hospital the following week.He was then started on bictegravir/emtricitabine/tenofovir alafenamide and a lumbar puncture confirmed the diagnosis of neuroinvasive JC virus, with a CSF DNA test of 2000 copies/mL . At our The patient was discharged without any changes in his neurologic symptoms. However, he returned the following month with a worsening gait disturbance, and new symptoms including dysarthria, dysmetria and left-sided nystagmus, despite a more than 3-log drop in his HIV RNA after 3 weeks of HAART , Fig. 2.The incidence of symptomatic PML infection increased significantly in the 1990s, as the AIDS epidemic expanded. Later, the disease was observed in those receiving novel immunosuppressive therapies for chronic inflammatory conditions The virus is thought to enter the body by inhalation or ingestion and can establish chronic infection in different tissues through its two forms: the archetypical (the transmissible), and the prototypical (the neuroinfective) form Our patient had two unusual features: first, he presented with lesions initially confined to the cerebellum and its reciprocal tracks. Second, his clinical course was not ameliorated following antiviral medication-induced control of his HIV infection.In cerebellar PML, the JC virus primarily targets granule cell neurons located in gray matter, which can lead to granule cell neuronopathy (GCN) IRIS associated to PML has been well described within the HIV population. His clinical worsening despite the control of his HIV infection was suggestive of IRIS When treating IRIS, rapid initiation of systemic steroids has demonstrated benefit in several co-infections In summary, our patient with newly diagnosed HIV and cerebellar JC virus infection worsened dramatically after introduction of effective antiviral treatment. We hypothesize that this may be related to the distinct pathophysiology of JC virus in the cerebellum as well as the deleterious effects of immune reconstitution.None to declare.N/A-case report.Written informed consent was obtained from the patient’s brother for publication of the details of their medical case and any accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal on request.Sergio Alvarez-Mulett: Involved in clinical care, data analyses and interpretation, drafting the manuscript and editing various drafts for critically important intellectual content, final approval of version to be published, agreement to be accountable for all aspects of work related to the accuracy or integrity for any and all aspects of the work and its review. Eli Sepkowitz: Involved in clinical care, data analyses and interpretation, drafting the manuscript and editing various drafts for critically important intellectual content, final approval of version to be published, agreement to be accountable for all aspects of work related to the accuracy or integrity for any and all aspects of the work and its review. Douglas Sepkowitz: Involved in clinical care, data analyses and interpretation, drafting the manuscript and editing various drafts for critically important intellectual content, final approval of version to be published, agreement to be accountable for all aspects of work related to the accuracy or integrity for any and all aspects of the work and its review.The authors have no conflicts of interest to declare. All authors contributed to the article and agreed upon the final document. No ongoing research, sponsorship or funding pertains to the present report.


Despite extensive research on biobased and fiber-basedmaterials,fundamental questions regarding the molecular processes governingfiber–fiber interactions remain unanswered. In this study,we introduce a method to examine and clarify molecular interactionswithin fiber–fiber joints using precisely characterized modelmaterials, i.e., regenerated cellulose gel beads with nanometer-smoothsurfaces. By physically modifying these materials and drying themtogether to create model joints, we can investigate the mechanismsresponsible for joining cellulose surfaces and how this affects adhesionin both dry and wet states through precise separation measurements.The findings reveal a subtle balance in the joint formation, influencingthe development of nanometer-sized structures at the contact zoneand likely inducing built-in stresses in the interphase. This researchillustrates how model materials can be tailored to control interactionsbetween cellulose-rich surfaces, laying the groundwork for futurehigh-resolution studies aimed at creating stiff, ductile, and/or toughjoints between cellulose surfaces and to allow for the design of high-performancebiobased materials. Nanocelluloses have been applied to createaerogels, nanopapers, energy storage devices, and responsive membranesjust to mention a few.2 Within many ofthese applications, the cellulose/cellulose interactions are imperativein order to utilize the inherent properties of the nanocelluloses.Cellulose-rich fibrous networksare ubiquitous in everyday life, both macroscopically in differentpackaging and hygiene products, but also in biocomposites and nonwovenmaterials that are used in a variety of products from simple wipesto medical dressing gowns.3 One obvious misconception is that hydrogen bondingis responsible for strong interactions at all length scales due tothe abundance of hydroxyl groups. Yet, considering the short-rangeand specificity of these interactions, this is a too simple and tooblunt description.4 One major reason forthe lack of fundamental data is the chemical and physical heterogeneityof cellulose-rich fibers, making them rather challenging for the determinationof molecular interactions. However, the development of smooth andchemically well-characterized model cellulose surfaces7 has paved the way for more fundamental studies of cellulose/celluloseinteractions. It was previously found10 that dispersive vander Waals forces greatly influence interactions between surfaces  under dry conditions, but in wet and moistconditions, swelling and capillary condensation begin to dominate.The large influence of water content during the preparation of cellulose-richmaterials on final materials properties has also been shown in a recentreview.11However, despite the obvious need for an understanding of the molecularinteractions between cellulose-rich surfaces, there is today no fundamentalunderstanding of the mechanism controlling these interactions.12 These investigations showed that there is alarge influence of the additives on the wet/drying properties of thecellulose surfaces.14 The inherent properties of theadditives (wet/dry) will significantly influence the interfacial structureand hence the molecular contact zone between the surfaces and naturallyaffect the overall final adhesion. These principal results are alsoin accordance with earlier studies of the fundamental interactionsbetween polybutylmethacrylate surfaces that were joined togetherunder different temperatures,15 indicatinghow the complex interphase would develop between cellulose surfaceswhen water is removed during drying.These investigations showthat the concept of explaining the interactionsbetween cellulose-rich surfaces by hydrogen bonding is an oversimplificationand that van der Waals interactions have a large influence on theinteractions. Moreover, the interactions between wet cellulose surfacesare also greatly affected by the swelling of the surface. It is alsoessential to determine the influence of how the water is removed duringdrying and how the mechanical properties of the outer layers of thesurfaces are affected by water removal. This later finding was alsosupported by rather recent investigations regarding the effect ofstrength-enhancing additives on cellulose and PDMS model surfaces.17 By using thismodel system, we have been able to study the fundamental interactionsbetween cellulose-rich surfaces during drying and how the separationforce after drying will be affected by treatments and testing conditions.These materials can then, under well-controlled conditions, be modifiedand characterized, dried together, and tested for adhesion, and finally,the contact zone between the materials can be analyzed both beforeand after separation.Taking this all togethermeans that in order to study the molecularinteractions between two cellulose-rich surfaces during drying, itis necessary to have access to initially smooth model cellulose surfaceswith controlled physical and chemical properties. During the lastyears there has fortunately been an interesting development of macroscopiccellulose surfaces with a spherical shape and a nanometer smooth surfaceallowing for these types of studies.Despite the wide use of strength additivesin fiber-based materials,it is unclear how, for example, a LbL assembly of polyelectrolyteson the surface of the fibers impacts the molecular interactions ofcellulose–cellulose joints. Moreover, to date, there has yetto be a demonstration of a model experiment that can accurately providemolecular-scale insight into cellulose–cellulose interactions.In this study, a model system using smooth macroscopic cellulose beadsis presented. The force required to separate the model joints withand without a well-defined LbL assembly of poly(PAH) and hyaluronic acid (HA) is evaluated. The separation stressis then determined by normalizing the force by the contact area determinedafter separation.Experimental section in SI).18 The resulting nanometer-smooth cellulosebeads, Cellulose beads were prepared by drippinga cellulose-LiCl/DMAc(lithium chloride dimethylacetamide) solution into a regenerationbath of ethanol  indentation techniques  for the indentation) and amethodology where the change swelling of the beads upon dissociationof the carboxyl groups was used to estimate the elastic response ofthe cellulose network in the beads. In the present work, this wascomplemented by a macroscopic elasticity measurement of the driedand reswollen beads using a specially developed methodology, describedin detail in the experimental section (in SI). These measurements are all representing different responses ofthe beads when they are dried together, and they are all importantfor the development of the molecular contact between the cellulosesurfaces. In Table SI.1, in Supporting Information, the results from all of these measurements are summarized. Thetrend for all measurements is the same, i.e., as expected, the modulusis decreased with the charge and, hence, water-induced swelling ofthe beads.To understand molecular interactions,it is essential to determinehow the charge of the beads will influence the wet modulus and ifand how additives affect these properties. In previous workTable SI.1,represent the elastic properties of the outermost surface of the beads,while the macroscopic modulus describes the response of the entirebead when compressed. Finally, the swelling response modulus showshow the cellulose network inside the beads responds when the chargesof the oxidized beads are dissociated. Previous investigations20 have demonstrated that the external and internalportions of cellulose fibers have a modulus of 0.01–1 and 0.1–5MPa, respectively, which are in fair agreement with the values in Table SI.1, indicating that the wet beads canserve as excellent models for the drying and consolidation of cellulose-joints.The AFM measurements, in 25 Here the LbL addition of poly (PAH) andhyaluronic acid (HA) onto model cellulose thin films was monitoredby stagnation point adsorption reflectometry (SPAR)26 to determine how much polyelectrolyte that is adsorbedto the surfaces. This is essential to allow for a fair interpretationof the collected adhesion resultsSurface modification and coatings can significantly alter the adhesionbetween cellulose joints. In this respect, the LbL assembly is a commonmethod used to controllably modify cellulose surfaces through thesequential addition of oppositely charged polymer or nanoparticlelayers.S/S0, increases with the addition of each polyelectrolyte.The adsorbed mass (mg/m2), as presented in 2627 In addition, the thickness of the dry thin filmswas measured with AFM .The macroscopic mechanical properties of the LbL-treatedbeadsare shown in 28 However,as was demonstrated earlier,29 it is also likely thatthe adsorbed PE layers also will have a wet structure, in additionto this drying-induced structure, that will significantly affect theadhesion between surfaces.While LbL adsorption has limited impact on the macroscopic mechanicalproperties, it is however expected that the adsorbed PEs will changethe surface structure of the beads. The surface morphologies of thedry, highly charged cellulose beads with and without surface treatmentwere imaged via AFM, and the results are summarized in The structure of the treated surfaces, as observedwith AFM 3,is furComparatively, the LbL-treated beads 4d–f are Before fully evaluating the adhesive propertiesof these joints,it is hence necessary to describe, in more detail, what is happeningupon drying two LbL-treated surfaces together. According to the AFMand SEM results, the LbL treatment, 5 bilayers, results in an around70 nm thick LbL structure on top of the cellulose surface. As thissandwich structure is drying, the slightly rougher LbL film will shrinkand wrinkle. Consequently, the structure of the cellulose surfaceis altered. This is schematically illustrated in Figure SI:1) than medium-charge beads and thus are more conformable and ableto form a larger contact area.In order to evaluate the stress at break when separating the beads,the contact zones were evaluated after separating the surfaces togetherwith the force at separation, and these results are shown in Figure SI.1) and thus could not form an effective joint when dried together.Low-charged (29 μeq/g)beads were significantly stiffer thanmedium- and high-charged beads , which all can be probed using the model cellulosejoints presented here.All this, hence, shows that the regeneratedcellulose beads canbe used as excellent model materials to investigate the mechanismsby which cellulose-rich surfaces form the initial contact and finallydry together to a joint and how surface modification impacts the formationand strength of the contact area. Specifically, by using smooth cellulosespheres, the surface structure and interface of the joints could bevisualized before and after separation. By sequentially adsorbingPAH and HA via a LbL assembly, a rough and structured surface wascreated, allowing for a more intimate contact at the interface. Theseadded layers influence the properties of the cellulose beads as wellas the formation of the contact zone between the surfaces during drying.Additionally, upon separation, the polymer chain disentanglement inthe wet/moist state leads to a high separation force and a large separationdistance of the LbL-treated cellulose surfaces due to the stretchingof the polymer complexes formed at the interface. These detailed insightswould not have been possible to elucidate without the technique presentedin this work.