The increased transmissibility of SARS-CoV-2 variants of concern (VOCs) has raised questions regarding the environmental stability of these viruses. Although a prolonged survival time has been reported for SARS-CoV-2, how long new variants can persist on contaminated surfaces and how environmental factors affect the persistence time are not fully characterized. The present study provides a comprehensive assessment of the stability of Omicron variants BA.1 and BA.5, which are currently circulating strains, on the surfaces of widely used transport packaging materials. By monitoring viable virus detection over a 7-day period under different environmental conditions, it was found that the environmental stability of SARS-CoV-2 Omicron variants depended heavily on the surface type, temperature, and virus concentration. In addition, virus nucleic acid exhibited high stability on the material surface independent of whether viable virus was detected. These findings provide useful information for logistics practitioners and the general public to appropriately deal with transport items under different conditions to minimize the risk of epidemic transmission.IMPORTANCE This study shows the environmental stability of SARS-CoV-2 Variants Omicron BA.1 and BA.5 on surfaces of widely used transport packaging materials. The findings demonstrate that the environmental stability of the SARS-CoV-2 Omicron variants varies based on material type. The viability of SARS-CoV-2 on material surfaces depends heavily on temperature and viral titer. Low temperatures and high viral titers promote virus survival. Moreover, in contrast to virus viability, virus nucleic acid exhibits high stability on the surfaces of widely used materials, making the detection of virus nucleic acid unsuitable for evaluating the risk of epidemic transmission. Since the first emergence of the SARS-CoV-2 virus in November 2019, various variants of concern (VOC) have emerged and spread globally. Among these VOCs, the currently circulating Omicron strain is a major public health concern owing to its high infectivity and transmissibility , were also included to precisely assess situations under different conditions. An understanding of the environmental stability and persistence of SARS-CoV-2 Omicron variants on contaminated surfaces has profound impacts on the handling of transport items.The virus viability of Omicron subvariants on the surfaces of four widely used transport packaging materials was investigated over time. There were three temperature conditions in this study: 4°C (low temperature), 25°C (room temperature), and 37°C (high temperature). At each temperature, 180 samples were recovered for each subvariant, including four material surfaces, five time points, and three RH conditions. At 4°C, 47 BA.1 samples and 68 BA.5 samples were detected with the viable virus ; at 25°CTable 150]), the viable virus of both subvariants was only detected at 6 h, and under very limited conditions, such as BA.1 on the surface of iron at 37°C with an RH of 80% (50), more positive samples were detected, and the time period of viable virus detection extended to 1 day under some conditions, especially at 4°C. With the high virus titer , an increased number of samples were positive for the viable virus for prolonged periods of time, and the persistence time of BA.1 and BA.5 on the surfaces of nonwoven fabric, PE packaging film, and iron extended to day 5 and day 7 at 4°C, respectively  and RH values of 40% and 60%. Its survival time on the surfaces of these three materials with an RH of 80% was reduced to 5 days (nonwoven fabric and PE packaging film) and 3 days (iron) (50) and a low virus titer (10 TCID50), the trend of an RH of 80% being more detrimental to the virus was only observed on 100 TCID50-treated paper carton and iron, and 10 TCID50-treated PE packaging film. Taken together, these results suggest that the role of humidity is more intricate, or that it coordinates with other factors.RH was also investigated as an environmental factor in this study. With RH values of 40%, 60%, and 80%, a total of 15, 16, and 16 samples were positive for viable BA.1, respectively, while 25, 25, and 18 samples were positive for viable BA.5 at 4°C during the tested period . With ths (iron) . However50 BA.5-inoculated nonwoven fabric at 4°C). The results showed that no discrepancy existed (P = 0.23)  , suggestContaminated fomites are thought to be a potential transmission route in the current pandemic caused by SARS-CoV-2 , 4. ThesThis study evaluated the environmental stability of two Omicron subvariants on the surfaces of four widely used transport packaging materials. Among these four materials, the Omicron variant was rapidly inactivated on the surface of paper carton, while it persisted for a longer period of time on the surfaces of nonwoven fabric, PE packaging film, and iron. These results are consistent with previous reports that SARS-CoV-2 has a long survival time on nonporous material or porous material made up of high-density hydrophobic ingredients, such as the outer layer of surgical masks and Tyvek , 5, 6. CThe environmental stability of SARS-CoV-2 has been compared with other viruses and between different SARS-CoV-2 VOCs –11. Thes–In this study, the effects of the temperature and RH on the stability of SARS-CoV-2 Omicron variants were investigated. The stability of viruses under different temperatures was consistent with previous studies that indicated that increases in temperature accelerated the inactivation of SARS-CoV-2 on surfaces , 13. HowOf note, while viable viruses decayed over an extended time, the virus nucleic acid showed high stability on material surfaces. In real-life settings, reverse transcription-PCR (RT-PCR) is widely used to determine the contamination of the virus in environment samples. However, it should be noted that a positive result of virus nucleic acid, even with low cycle threshold (CT) values, does not mean that infectious viruses exist. The results of RT-PCR should be interpreted and evaluated cautiously.In conclusion, this study demonstrates that the environmental stability of the SARS-CoV-2 Omicron variant varies based on surface type. The virus viability on surfaces was highly dependent on both the temperature and viral titer. Low temperatures and high viral titers promote the persistence of the virus. Moreover, in contrast to virus viability, virus nucleic acid has high stability on widely used materials, and is not suitable for evaluating the risk of epidemic transmission.The SARS-CoV-2 variants analyzed in this study were the Omicron BA.1 and BA.5 strains. The BA.1 virus (IPBCAMS-OM01-1/2021) was isolated from a clinical sample; BA.5 (CCPM-B-V-049-2207-28) was provided by the Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College. Vero cells (African green monkey kidney cells) were purchased from American type culture collection (ATCC) (CCL-81), cultured in Dulbecco’s modified Eagle Medium  supplemented with 10% fetal bovine serum (FBS), and used for virus culture and qualification. All work handling SARS-CoV-2 was performed in a Biosafety Level 3 (BSL-3) laboratory.2. PE packaging film was sterilized using 75% alcohol immersion. Other materials were sterilized via autoclaving. Pieces of materials were then placed in 12-well plates for inoculation with viral aliquots.The materials used in this study were widely used transport packaging materials provided by Beijing Customs, including paper cartons, nonwoven bags, PE packaging film, and iron drum packaging. All materials were cut into small pieces of about 1.5 cm50 according to our previous report  and incubated under various temperatures .The titers of Omicron BA.1 and Omicron BA.5 virus stocks were determined based on the 50% TCIDs report . The virThe virus was recovered from the surfaces of the materials at each time point by adding 2.0 mL DMEM to each sample. Then, 200 μL of DMEM collected from each sample was used for incubation with Vero cells. The cytopathic effects (CPEs) were observed 4 days later. If no CPEs were observed, the supernatants were collected and passaged three times. The observed CPEs in any generation were recorded to indicate the activities of the virus.Four hundred μL of each recovered sample was used for nucleic acid extraction with a Direct-zol RNA miniPrep kit  according to the manufacturer’s instructions. The virus nucleic acid copy numbers were measured using RT-PCR with primers and a probe targeting the SARS-CoV-2 N gene. Quantification was performed using the N target with a standard curve generated based on the serial dilution of the reference standard.


This study aimed to investigate the impact of TG5 (c. −422C > T) and LEP (c. 239C > T) polymorphisms and reared under the same fattening conditions. Controlled slaughter was conducted at 18 months of age. Subsequently, the experimental animals were evaluated to determine their slaughter traits and the chemical and FA composition of ground beef and the longissimus dorsi muscle.A total of 26 Simmental bulls were genotyped for TG5 (c. −422C > T) polymorphism significantly (p < 0.05) affected the differentiation of bulls in terms of the synthesis of stearic acid, linolenic acid, and total polyunsaturated FAs, as well as the fat and dry matter content in the longissimus dorsi muscle. Conversely, the presence of the T allele in the LEP (c. 239C > T) polymorphism was associated with increased dry matter and fat in ground beef, carcass weight, and internal fat weight.The results showed that the TG5 and LEP genes revealed a genetic basis for the quantitative and qualitative characteristics of meat productivity. Thus, the genetic variability of bulls regarding the LEP and TG5 genes can be used to improve the quantitative and qualitative indicators of meat productivity in Simmental cattle through marker-assisted selection.The analysis of slaughter traits and the chemical and FA composition of meat from the Simmental bulls genotyped for the  More than 80% of domestic beef production in Russia comes from slaughtering dairy and dual-purpose cattle , 2. Therleptin (LEP) and thyroglobulin (TG5) genes as significant genetic factors that influence the intensity and characteristics of fat metabolism in the body [C422T polymorphism occurs in the 5’-promoter region of the TG5 gene [LEP gene are extensively used in marker-assisted selection (MAS) programs to predict carcass fat formation, body weight, and growth rate in cattle [Beef, derived from cattle muscle and adipose tissue, undergoes various physical and chemical processes. The level and intensity of these processes, and hence meat quality, are determined by genetic and paratypic factors, such as temperature, storage time, sex, and age of the animal , 6. Red the body , 11. ThuTG5 gene . This nuTG5 gene , 14. Varn cattle , 16. DesTG5 (c. −422C > T) and LEP (c. 239C > T) polymorphisms.This study aimed to investigate the formation of meat productivity, chemical composition, and FA profile of beef in Simmental bulls based on The Local Ethics Committee of the Russian State Agrarian University-Moscow Timiryazev Agricultural Academy, Moscow, Russian Federation, approved the protocol of the present investigation . All animal studies were conducted following the ethical standards laid down in the 1964 Declaration of Helsinki and its later amendments. Every possible effort was made during the research to minimize animals’ distress and use fewer samples.The study was conducted from June 2021 to December 2022 at the test station of the “Dombarovskiy” Ltd. .th and 11th ribs.Simmental bulls (n = 26) with different genotypes were reared under the same fattening conditions up to 18 months of age. Animal-controlled slaughter was conducted following State Standard R 34120–2017, “Cattle for slaughter, Beef and veal in carcasses, half carcasses, and quarters” [The chemical and FA compositions of the ground beef and longissimus dorsi muscle were determined in the laboratory of the Department of Physiology, Morphology, and Biochemistry of Animals at the Russian State Agrarian University-Moscow Timiryazev Agricultural Academy. Dry matter content was determined by drying the samples in an oven at 100°C, while organic matter was determined by ashing the dried sample at 550°C . The FA TG5 (c. −422C > T) and LEP (c. 239C > T) gene polymorphisms, blood samples were taken from the jugular vein of the experimental bulls. DNA extraction was conducted using the DIAtom™ DNA Prep kit . GenePakPCRCore kits (IsoGeneLab) were used for polymerase chain reaction (PCR)-restriction fragment length polymorphism on a programmable “Tertsik” thermal cycler . Specific primers , PCR conditions, and endonucleases were used in this study .Model used:ij=μ+Ai+eij,Yij – represents the studied traits, μ – is the overall mean, Ai – is the fixed effect of the TG5 and LEP genotype , and eij – is a random error.Where, YThe significance of inter-genotype differences was assessed using a posteriori Fisher’s criterion (F-test). p ≤ 0.05 was considered statistically significant.LEP gene significantly impacted the variability of the slaughter indicators in Simmental bulls  and 0.5%, respectively. Heterozygous animals differed in their maximum indicators of carcass and slaughter yields, with the proportion of deposited fat at a minimum level. This finding indicates the relative late maturity of carriers of the CTLEP genotype. The T allele in the LEP (c. 239C > T) polymorphism was associated with increased live and carcass weight in Simmental bulls. However, when this allele was homogenized, fat accumulation significantly intensified with a slight decrease in weight growth and carcass weight relative to heterozygous peers.The polymorphism of the al bulls . HeterozTG5 gene polymorphism of Simmental bulls  polymorphism affected the chemical composition of the average ground beef sample (LEP gene polymorphism.Differentiation of bulls according to the f sample . The T aLEP (c. 239C > T) polymorphism. There was a tendency toward increased fat synthesis in muscle tissue in TT-homozygotes relative to CC-genotype by 0.46% (p = 0.16) and heterozygous individuals by 0.41% (p = 0.26). Conversely, homozygotes for cytosine tended toward more intensive protein synthesis, providing them with a 0.29%–0.49% (p > 0.05) advantage over their peers.The chemical composition of the longissimus dorsi muscle was influenced to a lesser degree by TG5 gene; however, intergroup differences were not significant .The advantage of heterozygous individuals in total nutrient accumulation in carcass flesh was observed when the bulls were genotyped for the nificant . The supTG5 (c. −422C > T) polymorphism had a more significant effect on the chemical composition of longissimus dorsi muscle compared to LEP gene. In particular, a highly significant superiority  of the heterozygous genotype was observed in fat accumulation in muscle tissue. There were less noticeable intergroup differences in protein synthesis. Thus, the muscles of heterozygous bulls contained 1.0% (p < 0.001) drier matter compared to their homozygous peers.The TG5 (c. −422C > T) polymorphism and the stearic (C18:0) acid content (CCTG5 genotype outperformed their heterozygous peers by 0.85% (p < 0.05) regarding the amount of this FA in the lipid structure. Intergroup differences were not significant for other SFAs, although heterozygous individuals had a trend-level advantage in the synthesis of myristic by 0.11% (p = 0.26) and palmitic by 0.55% (p = 0.58).The analysis of the FA profile of the longissimus dorsi muscle in Simmental bulls indicated a significant association between the  content . Carrier18:1), which varies within a narrow range of 38.75%–38.95% during genotyping for the TG5 gene. The intergroup differences were also insignificant for the other MUFAs. Thus, an advantage in the myristoleic (C14:1) acid content was observed among heterozygous individuals, amounting to 0.07% (p = 0.30), while homozygous individuals showed a 0.14% advantage (p = 0.29) in palmitoleic acid.The high content of monounsaturated FAs (MUFAs) in the lipid structure is associated with a low melting point of intramuscular fat. It positively affects the organoleptic characteristics of beef, particularly its taste and tenderness. The most important and representative MUFA in beef fat is oleic acid  polymorphism. Heterozygous bulls exhibited a significant advantage over their homozygous peers in the amount of linolenic (C18:3) acid by 0.07% (p < 0.05). In addition, they had a trend-level advantage in the synthesis of linoleic  and arachidonic  acids.The content of specific polyunsaturated FAs was largely determined by the allelic variant of the LEP (c. 239C > T) polymorphism had a less noticeable effect on the FA profile of beef from Simmental bulls  and 0.53%–0.88% (p > 0.05), respectively. Conversely, homozygotes for thymine exhibited an increased concentration of myristic (C14:0) FA, with an advantage of 0.05%–0.10% (p > 0.05) compared to other genotypes for the LEP gene.Our studies showed that the al bulls . HoweverHomozygous and heterozygous carriers of the T allele tend to increase MUFA accumulation in muscle tissue. They surpassed their peers in myristoleic and oleic acid content by 0.04% and 0.82%–1.13%, respectively.LEP (c. 239C > T) polymorphism was characterized by the best composition of polyunsaturated FAs. Carriers of the CTLEP genotype had an advantage in the proportion of linoleic and arachidonic acid by 0.11% and 0.08%, respectively, relative to their homozygous peers. Conversely, the TTLEP genotype was associated with an increased (by 0.04%) linolenic (C18:3) acid content.Meat from heterozygous bulls for the CCTG5 young animals slightly exceeded the SFA (0.19%) and MUFA (0.27%) content of their heterozygous peers  in polyunsaturated FA synthesis.Integral indicators of lipid metabolism largely characterize the meat quality of Simmental bulls. Thus, the homozygous us peers . In turnLEP (c. 239C > T) polymorphism also indicated the superiority  of heterozygous carriers in polyunsaturated FA synthesis  single-nucleotide polymorphism determined the variability of preslaughter live weight by 19.36% (p = 0.08), carcass weight by 17.30% (p = 0.11), internal fat weight by 14.63% (p = 0.16), internal fat yield by 12.65% (p = 0.21), slaughter weight by 20.12% (p = 0.08), fat content in ground beef by 26.01% (p < 0.05), and dry matter content by 38.02% (p < 0.01). Our findings were consistent with the results of Ardicli et al. [LEP gene on slaughter traits in Simmental animals. The contribution of the LEP gene polymorphism to the variability in the chemical composition of muscle tissue was slightly smaller. In contrast, the substitution C→T in the nucleotide sequence of the TG5 gene did not result in significant changes in slaughter indicators and the chemical composition of carcass flesh. Due to the low variability in protein content, differences in the chemical composition of ground beef were mainly due to the unequal intensity of fat deposition in carcasses from various genotypes of bulls when they were differentiated according to the LEP gene polymorphism, leading to the highest variability in the protein and fat ratio in carcass flesh (1:0.51–0.67). Conversely, the nucleotide substitution in the TG5 gene provided a variation of this parameter within 1:0.55–0.58.In our research, the variability of slaughter indicators and the chemical composition of carcass flesh in Simmental bulls, depending on the genotypes for the e LEP c. 9C > T siTG5 (c. −422C > T) polymorphism significantly (p < 0.001) explained the variability in fat content by 49.94% and dry matter content by 37.64% in the longissimus dorsi muscle. However, the content of major nutrients in ground beef showed low determination using the allelic structure of the TG5 gene, mainly due to the unequal distribution of alternative alleles in the herd. This allelic frequency limits selection for creating the optimal genetic structure in the population, considering the TG5 gene [et al. [The TG5 gene . Neverth [et al.  reportedet al. [et al. [CSN1S1 × TG5 genetic complex to increase the frequency of desirable alleles in the gene pool of an improved herd. Therefore, despite long-term purebred breeding, the Simmental breed retains significant genetic diversity that can further contribute to improvement through intrabreed resources [Shevkhuzhev et al.  recommen [et al.  proposedesources .TG5 (c. −422C > T) polymorphism compared to the allelic profile for the LEP gene. A similar conclusion was reached by Sedykh et al. [TTTG5 genotype carriers in terms of fat content in muscle tissue relative to their peers in Hereford bulls. However, they did not find significant differences between Hereford bulls of various genotypes regarding the LEP gene.Analysis of the chemical composition of the longissimus dorsi muscle in Simmental bulls showed a significantly greater determination of nutrient content depending on h et al. , who obset al. [et al. [LEP gene polymorphism, while bulls with the CTTG5 genotype, characterized by the smallest carcass weight, had the maximum amount of polyunsaturated FAs.De Smet et al.  reported [et al.  found thTG5 (c. −422C > T) polymorphism had a greater effect on the lipid composition of muscle tissue in cattle compared to the nucleotide substitution in the LEP gene. Specifically, differentiating Simmental bulls using the TG5 gene significantly determined the variability in the stearic acid , linolenic acid , and polyunsaturated FA  content. Carriers of the T allele showed increased synthesis of polyunsaturated FAs. A similar significant effect of TG5 gene polymorphism on the FA composition of blood serum was found in Hereford cattle [Our study showed that the d cattle . Consided cattle  and is cd cattle , the T aTG5 and LEP genes, revealed the genetic determination of the quantitative and qualitative characteristics of meat productivity. The TG5 (c. –422C > T) polymorphism significantly (p < 0.05) affected the differentiation of bulls in terms of the synthesis of stearic acid, linolenic acid, and total polyunsaturated FAs, as well as the fat and dry matter content in the longissimus dorsi muscle. Similarly, the T allele in the LEP (c. 239C > T) polymorphism was associated with increased levels of dry matter and fat in ground beef, carcass weight, and internal fat weight. Thus, the genetic variability of bulls in terms of the LEP and TG5 genes can be used to improve the quantitative and qualitative indicators of meat productivity in Simmental cattle using MAS.The analysis of slaughter traits, chemical composition, and FA profile of meat from Simmental bulls, genotyped for IS, EL, AM, OT, YK, DS, IB, and AO: Contributed equally to the experimentation. IS and EL: Wrote and edited the manuscript. AM, OT, and YK: Equally designed and conducted the experiment. DS, IB, and AO: Studied scientific literature about the topic. All authors have read, reviewed, and approved the final manuscript.


Magnetic beads (or particles) having a size between 1 and 5 µm are largely used in many biochemical assays devoted to both purification and quantification of cells, nucleic acids, or proteins. Unfortunately, the use of these beads within microfluidic devices suffers from natural precipitation because of their size and density. The strategies applied thus far to cells or polymeric particles cannot be extended to magnetic beads, mainly due to their magnetization and their higher densities. We report an effective shaking device capable of preventing the sedimentation of beads that are stored in a custom PCR tube. After the characterization of the operating principle, the device is validated for magnetic beads in droplets, leading to an equal distribution between the droplets, barely affecting their generation. Microfluidics is experiencing large diffusion in many different biological applications, especially in the development of a new generation of biomedical instruments devoted to the diagnosis of various types of cancer and degenerative diseases ,2. As a During recent decades, the use of magnetic beads has experienced a large diffusion in both research and clinical diagnostics, especially for the purification, isolation, and quantification of rare biomarkers, such as cells, nucleic acids, proteins, or extracellular vesicles ,11,12,133) dispersed in water  . These tThe shaking device shown in body see . In contbody see b was use3 and 1.05 g/cm3, respectively). Once the tube was turned upside down with respect to conventional use, the beads immediately moved above the oil  mounted on two independent syringe pumps  were used to control droplet generation and motion. The use of a syringe for the infusion of the bead suspension presented the drawback that part of the sample stored inside could not be completely flown into the microfluidic device, remaining in the syringe itself or along the capillary used for inlet connections to the microfluidic device. To avoid this problem, a conventional 0.5 mL PCR tube  was customized by fixing two capillaries  in two drilled holes: one on the side and the other on the cap, representing the inlet and the outlet, respectively see a,b. The  oil see . Thus, bDuring microfluidic experiments, PEEK capillaries were connected to PTFE capillaries  through a silicone joint .8 and 109 beads/mL). Before use, the magnetic beads were washed twice and stored in PBS  solution. Fluorinated oil  mixed with 2% surfactant  was used for droplet generation. The surfactant decreased the surface tension between the oil and the aqueous phase, allowing the generation of droplets and promoting their stability during motion [Aqueous suspensions of magnetic beads with a mean diameter of 2.8 µm  were prepared at different concentrations . In detail, polydimethylsiloxane  allowed replicating the initial brass mold, and the resulting part was then used as mold as well . SilanizTo acquire image sequences of the activation of the shaking device, a fast camera  was used, coupled to a macro zoom lens . Instead, a CMOS camera (Basler acA1300–200 µm) coupled with a telecentric lens  was used to evaluate the distribution inside the droplets after their formation within the microfluidic device and storage within the PTFE capillary. A white LED backlight was required to illuminate the capillary.The performance of the shaking device in preventing the sedimentation of the beads was validated by (i) measuring the bead sedimentation rate within the tube and (ii) by evaluating the magnetic bead distribution inside the droplets. In this second case, we also characterized the possible vibration effects on the droplet generation, in terms of the droplet size polydispersity.The shaking device was first characterized in terms of amplitude and frequency of vibration for different applied voltages (ΔV) to the motor, with the PCR tube filled with Phosphate Buffer Saline solution (PBS). Image sequences see  acquiredh) between the beads–oil interface and the upper tip of the PCR tube  and absence (static) of vibration. The difference between static and shaking conditions was also evident to naked eyes from the pictures reported in tube see a. The grTo evaluate the applicability of the shaking device in homogenously distributing magnetic beads over a sequence of generated droplets, the experimental setup reported in Consistent with the results of 2 in the graphs) was directly related to the quantity of beads in each droplet, as already applied for the immunoagglutination assay [8 beads/mL and 4 × 109 beads/mL). These values were chosen because they were typically used in many applications, such as immunoassays, mRNA and DNA purification, or extracellular vesicle isolation [9 beads/mL).Finally, to evaluate the magnetic bead encapsulation within the generated droplets, the latter was transported in correspondence with a magnetized iron tip, leading to clustering of beads see a. In paron assay . Figure solation ,46,47. FWe realized a shaking device that was effective for continuously homogenizing a suspension of magnetic beads within a storage tube. In this manner, the solution is ready to be used before and, more importantly, during automated microfluidic experiments that require a uniform distribution of beads for subsequent analysis purposes. The feasibility of our approach was effectively shown for a typical size of the magnetic beads (2.8 µm) at different initial concentrations and volumes. Furthermore, the shaking device could work for long-term experiments (tested up to 75 min). Finally, the inverted orientation of the PCR tube prevented the loss of sample, which, being collected from the narrow part of the tube, was completely injected within the microfluidic channels.The use of magnetic beads in droplet microfluidics is widely used in many biological applications  (e.g., aMagnetic beads are also employed in monophasic microfluidic devices for various biological procedures , such as


Recent studies have shown a lifetime prevalence of 5.7% for health anxiety/hypochondriasis resulting in increased healthcare service utilisation and disability as consequences. To the best of our knowledge, there has been no systematic review examining the global costs of hypochondriasis, encompassing both direct and indirect costs. Our objective was to synthesize the available evidence on the economic burden of health anxiety and hypochondriasis to identify research gaps and provide guidance and insights for policymakers and future research.A systematic literature search was conducted using PubMed, Web of Science, PsycInfo, EconLit, IBSS and Google Scholar without any time limit, up until April 2022. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed in this search and the following article selection process. The included studies were systematically analysed and summarized using a predefined data extraction sheet.Of the 3044 articles identified; 10 publications met our inclusion criteria. The results displayed significant variance in the overall costs listed among the studies. The reported economic burden of hypochondriasis ranged from 857.19 to 21137.55 US$ per capita per year. Most of the investigated costs were direct costs, whereas the assessment of indirect costs was strongly underrepresented.This systematic review suggests that existing studies underestimate the costs of hypochondriasis due to missing information on indirect costs. Furthermore, there is no uniform data collection of the costs and definition of the disease, so that the few existing data are not comparable and difficult to evaluate. There is a need for standardised data collection and definition of hypochondriasis in future studies to identify major cost drivers as potential target point for interventions. Hypochondriasis or, sometimes also referred to as health anxiety, can be broadly defined as the pathological fear of suffering from a serious physical illness, poses a major economic burden on healthcare systems and society. One of the most recent studies on the prevalence of hypochondriasis in the general population was conducted in Australia, and found a lifetime prevalence of 5.7%, with a peak in middle age . A systeThere seems to be no uniform definition for health anxiety and hypochondriasis. In the literature, these two terms are often used interchangeably, but some authors define them as two separate entities. The DSM IV  categorizes hypochondriasis as a somatoform disorder involving the misinterpretation of physical sensations . HoweverIn terms of therapy, cognitive behavioural therapy (CBT) is considered the most effective treatment for the disease and can also be offered as an online therapy . It is aHealth costs can be categorized into different types. There are the tangible, monetarily measurable costs. They differ in the cost perspective, i.e., whether they are incurred by the service provider, the patient, or society. Tangible costs are further classified into direct and indirect costs. Direct costs are expenses directly related to the disease, such as transport costs, expenditures for specialized medication, home modifications due to a disease, or therapies that the patient has to pay for out of pocket. If therapy or prevention is covered by health insurance, these direct costs are incurred by the service providers. Administrative costs, personnel costs, expenses for doctor’s visits and diagnostics, or research are also considered direct costs. Indirect costs are reflected in productivity losses, increased absenteeism from work due to sick leave and the associated loss of wages. Another type of costs are intangible costs, which will not be discussed in detail here .To the best of our knowledge, no existing systematic review has examined the economic impact of health anxiety and hypochondriasis on healthcare systems and societies at a global level. This paper aims to synthesize the available evidence on the economic burden of health anxiety and hypochondriasis, specifically focusing on total costs, including both direct costs of health anxiety/hypochondriasis caused by doctor visits, medication or administration and indirect costs resulting from absenteeism from work and productivity loss.As the first systematic review in this field, a key objective is to emphasize the disease burden, enabling resource allocation, healthcare prioritization, and identifying research gaps to provide guidance and insights for policymakers and future research.This systematic review was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) . It is rWe started a preliminary search to detect relevant search terms and develop a search string consisting of (i) the disease hypochondriasis and related synonyms and (ii) search terms relating to economic burden. The latter included the terms „health anxiety“, „hypochondria*“, and „illness anxiety disorder“. The second part included the terms „cost*“ and „economic“. An asterisk has been added to the terms cost and hypochondria to include other terms with the same root word in the search. Search terms within a category were linked with the Boolean operator „OR“ and then enclosed with brackets, the categories themselves where then linked with „AND“.We conducted the systematic literature search in the following databases: PubMed, Web of Science, PsycInfo, EconLit, IBSS and Google Scholar. No restriction on publication year was applied. In Google Scholar, we screened the results for each search combination to saturation which means we continued screening until no more relevant articles appeared, and our subject area was adequately covered. After reaching saturation, 30 more articles were screened. Saturation was achieved within a range of 50 to 180 articles depending on the search term used. Later, we screened the reference lists of the included publications to detect further relevant articles. The search was conducted in April 2022.All database hits were imported into Rayyan to remove duplicates and to screen the records . The titWe included any peer-reviewed study on the economic burden of hypochondriasis, such as direct costs, related doctor visits both primary and secondary care, medication, or administration as well as indirect costs resulting from absenteeism or productivity losses. Our systematic review included studies from low-, middle- and high-income countries. We encompassed all types of studies, including randomised controlled trials (RCT), quasi-experimental studies, observational studies, cohort studies, cross-sectional studies, cost-effectiveness, cost-utility, and cost-of-illness.We only included articles that reported costs incurred by health anxiety/hypochondriasis if the patients did not receive any special therapy like cognitive behavioural therapy. The rationale behind this criterion is that hypochondria still receives limited attention in diagnosis and treatment. Therefore, we aimed to showcase the costs that arise, particularly from untreated cases, to underscore the importance of adequate diagnosis and subsequent therapy for the disease. For comparative studies examining the cost of different forms of cognitive behavioural therapy, only the costs incurred by the control group, such as a waiting list control or treatment as usual (TAU), were included.There was neither an age limitation of participants nor a date restriction regarding the publication date. Consequently, the included population consists of adults, young adults, and children. Included publications had to be indexed in English language.We did not consider studies that analysed the cost of cognitive behavioural therapy (CBT) as well as internet-guided CBT (iCBT) as treatment for health anxiety. For better comparability and to examine the total costs objectively, we excluded studies that did not provide monetary values of incurred cost items, such as the frequency of doctor visits or number of days absent from work. Due to low quality of evidence, we did not include case reports, narrative reviews, expert opinions, or editorials.The included studies were subjected to systematic analysis, and the relevant information was summarized in a predefined data extraction sheet. This data extraction sheet includes general information such as the title, author, journal, date of publication, country, study type, study objective, study setting, source of data, diagnostic assessment tool, definition of the disease used, and the time frame of the study. Additionally, the characteristics of the study participants, such as the number of participants, gender, and age, were analysed. Regarding our primary outcome, the total costs, as well as the different types of costs, were collected along with the corresponding currency. As reported costs related to different years, currencies and countries, all cost information was converted to US$ for the year 2022 using the CCEMG - EPPI-Centre Cost Converter .We assessed the quality of included studies using the reporting guidelines suggested by the equator network . In the In total, our database search in 2022 yielded 3044 articles. After the duplicates were removed, 19 articles remained for full-text screening. The screening of references of included articles did not reveal any further studies.Since three articles represented secondary analyses of already included studies, data of those studies were no longer considered. Four studies were excluded as there were no costs examined and one study was not peer-reviewed. Another study only looked at the costs of CBT without a control group. Finally, 10 studies met the inclusion criteria and were included in the systematic review. , which all can be classified as high-income countries according to the World Bank income group classification . AltogetTwo studies were non-interventional studies that compared the costs caused by hypochondriasis with the costs incurred by participants with other well-defined medical conditions , 19. SixWhile eight of the studies focused on adult participants suffering from health anxiety/hypochondriasis , 24–26, All studies contained information on the sex distribution of the study participants, either in the form of percentages, absolute values, or both –26. One For detailed information see Table Most of the included studies used the terms health anxiety and hypochondriasis interchangeably and employed similar assessment tools to identify the study participants , 24–26. The earliest study examined both hypochondriacal health anxiety and somatisation. The patients were given a questionnaire inquiring about somatic symptoms and hypochondriacal fears. Based on a predefined cut-off, the patients were classified as being above or below the somatisation threshold  . In DenmFive of the included studies examined the medication costs. To enhance comparability, these costs were also converted to annual costs in US$ for the year 2022. In Denmark, this resulted in costs of 400,13 US$ per year regarding DSM-IV diagnosis . In GreaAs already mentioned, indirect costs were also examined in 3 publications. These are also made up of various cost components. In all studies, however, the cost item sick leave was examined. This resulted in annual costs of 419.67 US$ and 1432.60 US$ in Sweden and 1212.65 US$ in Denmark , 20, 24.To our knowledge, this is the first systematic review to examine the total costs of health anxiety/hypochondriasis. Our aim was to summarize the available evidence on the costs of hypochondriasis, especially when patients are not receiving treatment, in the form of CBT or other treatment approaches. We were able to identify research gaps and draw implications for future research and policy. This systematic review revealed a lack of data about indirect and direct costs of hypochondriasis. Research on this topic has only been conducted in the United States and 3 different countries across Europe. All of them are high income countries. There were large cost differences observed not only between but also within the countries investigated. The 12-month costs ranged from 857.19 to 21137.55 US$ –26.There are several factors that contribute to this heterogeneous data situation. Firstly, the studies did not use standardised definitions and classifications of health anxiety/ hypochondriasis. As previously mentioned the definition of this condition has been subject to major discussions in recent years . In partWhile some studies consider all participants above a certain cut-off point  to be equally affected by health anxiety , 25, 26,Additionally, different cost items were included in the studies and the type of data collection also influenced the type of cost detail.This is a widespread issue in health economic studies. A systematic review conducted in 2021 compared various guidelines for health economic evaluations and revealed a substantial overall variability in determining which costs should be included .While especially in Denmark national health registers were used , 21–25, For the control groups receiving standard care, the knowledge that the participants suffer from hypochondriasis may have influenced the costs. The information about the disease can be viewed as a form of mini-intervention, as the confrontation with one’s own illness may have a therapeutic effect. In such cases, the costs would be lower than expected.Except for one study, all surveys were conducted in tax-funded health systems with no out-of-pocket payments. This could potentially impact resource utilization and, consequently, costs. However, it can also be inferred that the findings from these studies may not be generalizable to countries with different healthcare systems, particularly low- and middle-income countries.A systematic review that examined the global economic costs of anxiety disorders has revealed that indirect costs account for a significant proportion of the total costs . Since iResearch about hypochondriasis in children appears to be underrepresented. Only two studies, both conducted by the same team within the same cohort, attempted to gain further insights into the economic burden of the disease in this age group , 23.One reason for the missing data could be that diagnostic criteria for hypochondriasis and somatoform disorders for children are still not established. This problem was already identified 25 years ago . The ChiThis systematic review is subject to several limitations. Firstly, only articles indexed in English were considered which might have caused a lack of information about cost in non-Englsih speaking countries. Secondly, a limited number of studies were identified for inclusion, and the included studies often contained small study populations. Moreover, the heterogeneity in study design and data collection methods suggests that the results may not be generalizable. Furthermore, it is important to emphasize that the data cannot be generalized, particularly to non-high-income countries due to missing data. The variations in disease definitions and included cost components make data comparability challenging. Lastly, as only total costs were analysed, it is not possible to draw conclusions on specific cost drivers from this research.This systematic review offers valuable insights into the costs related to hypochondriasis while identifying relevant research gaps. However, it is important to note that the costs presented in this review are likely underestimated. This is primarily due to the variation in study methodology, where most studies only reported direct costs and employed different definitions of hypochondriasis or health anxiety, including various cost components. Establishing a comprehensive health care system with expanded therapy, prevention, or rehabilitation services relies on solid evidence. For this purpose, a guidance for a standardized and itemized cost survey is essential to identify potential cost drivers that can serve as targets for interventions. Future research especially for policy making should therefore seek to ascertain indirect costs on one hand, as it is likely that these add a significant burden to overall costs. On the other hand, we observed that there is a lack of uniformity of data collection and diagnostic criteria for the condition. Therefore, it is crucial to conduct further studies with consistent data collection and disease definitions.To be able to assess the costs in a global context, low- and middle-income countries should also be the subject of further research, as only high-income countries have been considered so far. Furthermore, it is important to conduct research that examines the impact of health anxiety on the gross domestic product and the proportion of the total healthcare costs to underline the importance of appropriate interventions and possible savings through them. This could be particularly relevant as an increase in health anxiety in the form of cyberchondria as already explained in the introduction can also be expected in the future due to increasing digitalisation .


We detected highly pathogenic avian influenza A(H5N1) clade 2.3.4.4b virus in a domestic cat that lived near a duck farm infected by a closely related virus in France during December 2022. Enhanced surveillance of symptomatic domestic carnivores in contact with infected birds is recommended to prevent further spread to mammals and humans.  On December 27, 2022, the avian influenza National Reference Laboratory of the Agency for Food, Environmental and Occupational Health & Safety in France confirmed a case of highly pathogenic avian influenza (HPAI) A(H5N1) clade 2.3.4.4b virus in a domestic cat. The cat lived with a human family next to a breeding duck farm, which had notified the animal health services of possible HPAI on December 9, 2022, after observing a 20% drop in egg production. After HPAI H5N1 clade 2.3.4.4b virus was confirmed at the farm, 8,375 ducks were culled on December 14. On December 20, the cat displayed disturbances in general condition, including apathy and mild hyperthermia, and was seen by a veterinary surgeon. The cat's condition worsened; pronounced neurologic and respiratory (dyspnea) symptoms appeared, resulting in compassionate euthanasia on December 24. Veterinarians collected sinonasal, tracheal, and anal swab samples after death, and a screening laboratory performed real-time reverse transcription PCR (RT-PCR) targeting the matrix protein and hemagglutinin (H5) genes. The laboratory sent H5-positive tracheal and sinonasal swab samples to the National Reference Laboratory, which confirmed HPAI H5N1 virus by using specific real-time RT-PCR for H5 clade 2.3.4.4b and neuraminidase (N1) genes .https://www.gisaid.org), 20 of which have the E627K mutation, most probably indicating a rapid selection of this mutation in mammalian hosts (We compared the complete sequence of the HPAI H5N1 virus found in the cat (A/cat/France/22P026544/2022) with other HPAI H5N1 virus sequences circulating in France in the same area, including the virus found in the neighboring duck farm (A/duck/France/22P025647/2022). Phylogenetic analyses of HPAI H5N1 genomes indicated that the virus from the cat belonged to the A/duck/Saratov/29-02/2021–like genotype, which has been the predominant virus genotype circulating in France and Europe since September 2022. The cat virus sequence was directly related to virus sequences identified in the same area in December 2022 . FurtherSince winter 2021–22, the number of reported cases of HPAI H5N1 clade 2.3.4.4b infections in mammals has increased  clade 2.3.4.4b virus in domestic cat, France, 2022.


Since May 2022, cases of monkeypox have been reported in around 70 countries where the disease is not endemic. The World Health Organization (WHO) has declared the 2022 monkeypox outbreak a Public Health Emergency of International Concern. The monkey poxvirus, like that of smallpox, is a member of the Orthopoxvirus group. There are two distinct strains  of monkeypox: the West African strain and the Congo Basin strain . ParticuIn the latter case , a brushConclusion: given the lack of specific strategies for the management of oral monkeypox virus infections, current evidences in the literature suggest that PDT seems to be a promising therapeutic approach, also by virtue of the absence of side effects and the low therapeutic cost. Therefore, although more studies are needed in this regard, in the light of the results that have emerged.


Generative design is a system that automates part of the design process, but it cannot evaluate psychological issues related to shapes, such as “beauty” and “liking”. Designers therefore evaluate and choose the generated shapes based on their experience. Among the design features, “complexity” is considered to influence “aesthetic preference”. Although feature descriptors calculated from curvature can be used to quantify “complexity”, the selection guidelines for curvature and feature descriptors have not been adequately discussed. Therefore, this study aimed to conduct a systematic classification of curvature and a feature descriptor of 3D shapes and to apply the results to the “complexity” quantification. First, we surveyed the literature on curvature and feature descriptors and conducted a systematic classification. To quantify “complexity”, we used five curvatures  and a feature descriptor (entropy of occurrence probability) obtained from the classification and compared them with the sensory evaluation values of “complexity”. The results showed that the determination coefficient between the quantified and sensory evaluation values of “complexity” was highest when the mean curvature was used. In addition, the Casorati curvature tended to show the highest signal-to-noise ratio . These results will foster the development of generative design of 3D shapes using psychological evaluation. In recent years, computer-aided automatic design technology, referred to as “generative design”, has been put into practical use and applied to product design. Generative design is defined as a design system that automates part of the design process  or utiliThe advantage of generative design is that it enables the generation of shapes that satisfy physical characteristics such as stiffness and mass. However, its shortcoming is that psychological characteristics such as “beauty” and “preference” of the shape, which are important in the early process of a product design, are not evaluated. Therefore, designers need to evaluate and select shapes on the basis of their experience and intuition . SolvingWhen evaluating a design, designers tend to focus on macroscopic shape features, which are defined as those that appear in the entire shape as a result of combining points, lines, surfaces, and other modeling elements . The maiRecent studies have indicated that a relationship exists between curvature and complexity ,17,18. TThis study aims to perform a systematic classification of curvature and feature descriptors of 3D shapes and to apply the results to a method for quantifying complexity. This paper is organized as follows. This section describes the systematic classification of curvature and feature descriptors of 3D shapes. We conducted a systematic classification according to a protocol known as preferred reporting items for systematic reviews and meta-analysis (PRISMA) . One feaA paper search was conducted using the “Web of Science” database. Specifically, the search criteria were set to include all keywords related to “curvature”, “3D”, “shape”, and “shape features”, which were relevant to our study. Papers evaluating 3D shapes were extracted. That is, we excluded both papers based on two-dimensional (2D) shapes and papers based on silhouettes or images of 3D shapes.Among the references extracted in procedure 1, we selected those that evaluate shapes via, for example, classification, retrieval, and recognition based on the features extracted from the shapes. Our reasoning is that these references are highly relevant to the objective of the present study, which is to evaluate the complexity of shapes. Other references involving the generation/deformation of shapes, such as by meshing, rendering, or noise detection, were eliminated.Among papers extracted in procedure 2, we selected those that evaluate macroscopic features by calculating a feature descriptor for each shape, such as similarity or complexity, because these references are highly related to evaluations of the degree of complexity.To narrow the papers obtained in the previous subsubsection to the literature highly relevant to the present study, we carried out a paper selection process based on titles and abstracts.Curvature is calculated from a 3D shape;A feature descriptor is calculated on the basis of the curvature.From the papers selected as described in Papers that did not satisfy both criteria, such as those involving calculations of a curvature or feature descriptor, were excluded from our survey.We obtained 1310 papers as a result of keyword settings. The shapes to be evaluated were classified into 3D and 2D shapes (generated from silhouettes or images of 3D shapes).The objectives of the studies on 3D shapes were divided into three main categories: (1) shape evaluation, (2) shape generation, and (3) shape deformation. Shape evaluation is the process of extracting features from shapes and evaluating their characteristics . Shape generation refers to the generation of 3D shapes by rendering or mesh segmentation of data such as 2D images or point clouds. Shape deformation denotes the process of removing points that deviate from the actual shape by detecting noise in the point cloud acquired by 3D scanning.The tasks for shape evaluation were further classified into multiple shape evaluations  and single shape evaluations .The details of the paper selection procedure are described below. First, 46 papers were selected on the basis of titles and abstracts. In the present study, following the criteria described in In these papers, polygon models were used to represent 3D shapes. A polygon model is a method of representing 3D shapes using a set of minute polygons, such as triangles and quadrilaterals. Because polygon models contain information about the shape surface necessary for calculating curvature and are commonly used in the field of industrial design ,24, polyIn the 18 extracted papers, seven types of curvature  were used. ry point .A feature descriptor that uses the curvature of each vertex of a polygonized shape is a vector whose elements are the curvatures at all vertices . CompareA feature descriptor that uses the curvature of each vertex and its surrounding vertices is a vector whose elements are calculated from the curvature of each vertex and its surrounding vertices, or it is a scalar obtained by their calculation  . CompareA feature descriptor that uses occurrence probabilities is a vector whose elements are the occurrence probabilities of curvature within a set range of curvatures, or entropy calculated from the vector . CompareA feature descriptor that uses transition probability is a vector of transition probabilities (probabilities of changing from one curvature to another) between two adjacent vertices within a set curvature range, or the entropy calculated from this vector, similar to occurrence probability . CompareA feature descriptor that uses clustering methods  can reduce the information amount by clustering vector-type curvatures such as the cone curvature and curvature tensor to clarify the remarkable vertices of the shape. This descriptor cannot be used for scalar curvatures.In the 18 extracted papers, five types of feature descriptor were used. This section describes a method for quantifying the complexity of 3D shapes. They have a large computational load to calculate the curvature. Both curvatures are expressed as an array of curvatures for a single vertex. Therefore, they are computationally more expensive than other curvatures where a single value is calculated for a single vertex. In addition, the computation required to compute feature descriptors from curvatures can also be substantial.They are difficult to use to evaluate curvatures of different sizes. A region  must be set as a parameter. However, in actual design situations, many scenarios arise where shapes of different sizes are evaluated, and setting a common region is difficult.This subsection describes the calculation methods of the curvatures in The following illustration of the curvature calculation is in accordance with the polygon mesh in ith vertex Using Gauss–Bonnet’s law and paraboloid fitting, Gaussian curvature equation :(1)Ki=2πith vertex Using Gauss–Bonnet’s law and paraboloid fitting, mean curvature  follows :(2)Hi=14The mean curvature is the curvature defined by the average of the principal curvatures, whereas the Gaussian curvature is the curvature defined by the product of the principal curvatures. Therefore, principal curvatures C can be calculated using the principal curvatures obtained from Equations (3) and (4):The Casorati curvature SI can be calculated even for a flat surface by dividing the cases according to whether The shape index can be calculated using the principal curvatures obtained from Equations (3) and (4). However, such an equation cannot be used to calculate the shape index in the plane CI can be calculated even for a flat surface by dividing the cases according to whether The curvature index can be calculated by using the principal curvatures obtained in Equations (3) and (4). However, the resultant equation cannot be used to calculate the curvature index in the plane Sukumar et al. , who invThe feature descriptors that consider the surrounding information are excluded because the computational load to integrate the curvature of each vertex and its neighboring vertices into a single value by calculation is larger than that of the other descriptors. In addition, the purpose of considering surrounding information is to evaluate the similarity of shapes accurately, not to evaluate human cognition.Feature descriptors that consider transition probabilities are excluded because evaluating the surface features of the entire shape is difficult. Although the feature descriptor can evaluate the transition of the curvature from one vertex to another, people are unlikely to focus only on local changes between vertices when evaluating complexity. In addition, checking and considering the results based on graphs such as contour plots are difficult when using transition probabilities.Feature descriptors that consider clustering are excluded because evaluating the curved surface features of the entire shape is difficult. The feature descriptor extracts salient points (vertices with characteristic curvature) from the entire shape as local features to evaluate the similarity among shapes deformed in only one part. However, curvatures not considered salient (not complex) are not reflected in the feature descriptors. In addition, as mentioned in the previous section, clustering can only be applied to a vector curvature , where multiple values are calculated from a single vertex.Feature descriptors that consider surrounding curvature information, those that consider transition probabilities, and those that consider clustering were excluded from our calculation. Our reasoning is described below.In this subsection, we first provide an overview of information entropy and its calculation method. We then outline the method used to calculate feature descriptors using the probability of occurrence in the present study.Information entropy equation : (8)E=−∑The curvature at each vertex of the 3D shape to be evaluated is calculated as described in The curvature at each vertex is discretized. Specifically, the minimum area value After discretization, occurrence probability The entropy of the probability of occurrence is calculated using Equation (8).This section describes the sensory evaluation experiments conducted to analyze the relationship between the feature descriptors calculated from five curvatures and sensory evaluation values of complexity. We first describe the experimental method, then present the results and discuss the sensory evaluation experiments conducted on three types of shapes.In this experiment, extruded and rotated shapes, which are typical CAD shape deformation methods, were used, along with the shapes of an actual product as the “complexity” evaluation sample. The details of the shapes are described below.Extruded shapeExtruded geometry is a 3D curved surface created from closed planar curves using extrusion, which is a 3D CAD geometry creation method. The extruded geometry was chosen for evaluation in this study because it enables the curvature characteristics of the planes, contours, and sides of an extruded surface to be compared. In this experiment, 50 2D closed curves used by Ujiie et al.  were ediRotated shapeA rotated shape is a three-dimensional curved surface created from closed planar curves using rotation, which is a shape creation method of 3D CAD. Rotational geometry was chosen as the target of evaluation in the present study because it enables the differences in curvature among planar, convex, and concave surfaces to be considered.The rotated shape used in this experiment was created by editing 50 closed planar curves similar to the extruded shape in 3D CAD and rotating the right side of the shape 360 degrees using the vertical direction passing through the center of gravity of the 2D shape as the rotation axis b. The siVase shapeA vase is a product with a high degree of freedom in its shape, and various types of curved surfaces can appear. Therefore, using the vase shape as the evaluation target, we can consider the influence of various curved surfaces on the perception of complexity. In addition to its functional role of holding cut flowers, a vase also plays a role as an interior decoration; thus, it is important to take aesthetic preferences into account when modeling . A vase A total of 50 vase shapes were used in this experiment: 25 vase shapes provided by Free3D  and 25 vEvaluation method: A 5-point Likert scale was used for complexity: “low complexity: 1”, “somewhat low complexity: 2”, “undecided: 3”, “somewhat high complexity: 4”, and “high complexity: 5”.z-axis: 9.0 s per rotation).Presentation method: An online questionnaire was used, with the sample shape colored gray and displayed on a white background. In addition, 10 of each sample shape were randomly selected and presented in duplicate to check the accuracy of the participants’ evaluation. Each shape was rotated at a constant speed  from uncomplex surfaces  is consistent with the human perception of complexity and is considered to have a high determination coefficient. Similarly, in the extruded and vase shapes, the number of states that discriminate complex and uncomplex surfaces tends to have a high determination coefficient c,d.2.Mean curvatureAs an example, we compare the number of states 7 with 6, where the change in the determination coefficient is large in the extruded shapes. In the extruded shapes, where the entropy variation is particularly large, the surface is discretized for each number of states as follows :For ed areas a.For ue areas b.The experimental results show that the Casorati curvatures are stable and highly correlated for all the investigated shapes. However, depending on the number of states, the determination coefficients of the Gaussian curvature and mean curvature can exceed the determination coefficient of the Casorati curvature. Therefore, we discuss the factors by comparing the determination coefficients of the Gaussian curvature and mean curvature with those of the Casorati curvature at the number of states where the determination coefficients of the Gaussian curvature and mean curvature are maximal. Gaussian curvatureAs an example, in the extruded geometry, the highest determination coefficient when using the Gaussian curvature is found for a number of states 18. We focus on the extruded shape, where the entropy change between the two curvatures is large. At each curvature, the surfaces of extruded shape are discretized as follows .For the Gaussian curvature, moderate surfaces with a principal curvature of 0 at one side and flat surfaces are discretized into gray states.For the Casorati curvature, moderate surfaces with a principal curvature of 0 at one side are discretized into different states: blue, gray, and red.Unlike the Gaussian curvature, the Casorati curvature can take positive values in the column plane. In addition, compared with the Gaussian curvature, the Casorati curvature tends to be larger for small (near zero) changes in principal curvature. Therefore, for extruded shapes, the Gaussian curvature is expected to quantify complexity more accurately than the Casorati curvature.2.Mean curvatureFor the mean curvature, flat and principal curvatures of 0 at one side are discretized into gray states, whereas convex edges are assigned to the red area and concave edges to the blue area.For the Casorati curvature, flat surfaces and concave surfaces are discretized into the same blue area. Convex surfaces as edges are assigned to gray or red states.As an example, for the vase shape, the highest determination coefficient when using the mean curvature is found when the number of states is 19. We focus on the vase shape, where the entropy change between the two curvatures is large. For each curvature, the surfaces of the vase shape are discretized as follows :For the Vase shapes tend to have a higher complexity evaluation when both convex and concave surfaces are included. Therefore, mean curvatures with different states for convex and concave surfaces are considered appropriate.In the present study, the entropy of occurrence probability was calculated as a feature descriptor because it can be used to evaluate the diversity of curvature and because the discretization parameters can be set according to human cognition. To verify the validity of this feature descriptor, we calculated the curvature moment as a comparison target.The curvature moment is a characteristic value calculated by considering the curvature group calculated from the shape as a probability distribution. Because some studies have evaluated the similarity of shapes using moments of curvature , it is aThe lower determination coefficient when using the curvature moment is attributed to the effect of mesh irregularities. In this subsection, we focus on the standard deviation that exhibits the highest desirability characteristic SN ratio among the curvature moments. The shape shown in One limitation of this study is that only the determination coefficient of the logarithmic approximation between the quantified and sensory evaluation values of complexity was used to evaluate their correlation. Although we confirmed some scatter diagrams  and thisWe conducted a systematic classification of curvature and feature descriptors in 3D shapes. The results showed that Gaussian curvature, mean curvature, Casorati curvature, curvature index, shape index, curvature tensor, and cone curvature can be used as curvatures calculated for 3D shapes. In addition, we confirmed that “feature descriptors considering surrounding information”, “feature descriptors considering occurrence probability”, “feature descriptors considering transition probability”, and “feature descriptors considering clustering” could be used as feature descriptors calculated for the 3D shapes.We applied the results of the systematic classification to a method for quantifying the “complexity” of 3D shapes. Specifically, Gaussian curvature, mean curvature, Casorati curvature, curvature index, and shape index were calculated for the polygon model. In addition, the probability of occurrence (information) entropy of these five types of curvatures was calculated as a feature descriptor.Finally, we conducted sensory evaluation experiments to verify the validity of the quantification method; the results showed that the highest determination coefficient for all three shapes was obtained when the mean curvature was used. To investigate the effect of discretization when calculating entropy, we calculated the mean value, standard deviation, and the SN ratio of the determination coefficient when the number of states was varied. The results showed that the Casorati curvature was stable for almost all of the investigated shapes.Our future work will include (1) conducting sensory evaluation experiments using other sample shapes to verify whether the same trend can be confirmed and (2) constructing a comparative criterion to enable a comparison that includes other types of curvatures and feature descriptors excluded in the present study.


Corvus corone cornix). West Nile virus (WNV) lineage 2 was identified for the first time as the etiological agent responsible for the death of the birds. WNV was detected in 17 out of the 99 (17.17%) free-living birds tested in this study. All the WNV-infected dead birds were collected in the same area and were diagnosed in September by the NVRI and confirmed by the EURL for equine diseases, ANSES, in October 2022. Unnaturally high temperatures recorded in Poland in 2022 likely favored the infection and spread of the virus in the avian population. A nationwide alert and awareness raising of blood transfusion centers and hospitals was carried out to prevent human infections by WNV.The summer temperatures recorded in Poland in 2022 were among the highest in over 30 years and, combined with higher-than-expected rainfall, gave the impression of an almost tropical climate. Such climatic conditions were ideal for the transmission of vector-borne zoonotic diseases such as West Nile fever. In northeastern Poland, in the Mazowieckie region, the Polish event-based surveillance network reported increased fatalities of free-living hooded crows ( Flaviviridae family, genus Flavivirus, which circulates in an enzootic transmission cycle between mosquitoes mainly from the Culex genus and wild bird species as an amplifying host, by mortality and viremia, and wild bird species consequently play a relevant role as reservoirs [West Nile virus, an enveloped RNA arbovirus, is a member of the servoirs ,2. Humanservoirs ,4,5. WNVservoirs  and has servoirs . WNV infservoirs ,9.Anas platyrhynchos) [Free-living predatory birds remain a perfect reservoir for a number of viruses that are commonly contagious and fatal for domestic birds but persist in the subclinical stage without any signs of clinical disease. WNV infections have been documented in many taxonomic groups of captive and wild birds , such ashynchos) . As evidhynchos) . WNV-neuhynchos) .Culex mosquitos, such as Culex modestus [Culex pipiens [Poland is located within central migration routes between Africa, Europe, and Asia and is home to many bird species as well as an important migratory stop for birds. Climate change could alter the migration corridors as well as the life cycle of modestus  and Cule pipiens ,15, caus pipiens . Such a  pipiens . Even th pipiens ,19,20. I pipiens  such as  pipiens  and the  pipiens .Anseriformes, Passeriformes, Strigiformes, Falconiformes, Ciconiiformes, and Accipitriformes orders were reported by the Polish event-based surveillance system. We expected transmission in autumn, connected with the life cycle of mosquitoes of the genus Culex, which are the main vector of the virus, and with the migration of birds, which are the main reservoir of the virus.During the 2022 transmission season, between 11 September and 28 October 2022, over 99 fatal cases among free-living birds belonging to The aim of this study was to confirm whether the deaths of these birds were related to viral infection and especially to the West Nile virus. We also evaluated the available data concerning WNV infection in free-living birds in Poland.The studies were conducted in Warsaw , a region in the central part of the largest city in the country and in the Central Mazowieckie Lowlands, on the Vistula River, Poland . The cliAll samples tested in the studies were obtained from the “Bird Asylum” Rehabilitation Centre for Protected National Birds (ptasiazyl.zoo.waw.pl) and from official veterinarians following handling procedures implemented following European (86/609) and Polish laws  with the aim to reduce the suffering of animals. The research in the above manuscript was performed on the carcasses of dead birds in accordance with the rules of ethics.From September 2022 to October 2022, 99 free-living birds belonging to 6 different orders represented by 10 avian species were collected in collaboration with the “Bird Asylum” Rehabilitation Centre for Protected National Birds and official veterinarians. Brain samples were collected during the necropsy of each individual bird. Among the 99 wild birds, 21 presented with neurological signs such as tremors, unnatural behavior with abnormal body posture, seizures, and paralysis, as reported by the veterinarians of the Rehabilitation Centre for Protected National Birds and official veterinarians.Viral RNA was extracted from 200 µL of brain homogenates using the viral RNA Mini Kit  following the manufacturer’s protocol. Viral RNA was suspended in an RNase inhibitor buffer , and concentrations were determined using a spectrophotometer . Viral RNA was kept at −80 °C before use.The WNV infectious strain New-York 1999 (GenBank accession: AF196835) isolated from the brain of an infected horse was kindly provided by the European Union Reference Laboratory for WNV studies . Two negative controls were used during RT-PCR and were acquired from chicken embryo fibroblast (CEF) and SPF cultures.Reverse transcription RT-PCR primers were designed to target the conserved 3′NCR sequence , and the sequences were as follows: WNVF (sense primer): 5′ AAA GCC CAA TGT CAG ACC AC 3′ and antisense primer WNVR: 5′ TAG TCC TTT CGC CCT GGT TA 3′, as indicated and developed by Niczyporuk, 2011.The RT-PCR method described by  was emplSequencing libraries were prepared from two pools of genomic RNAs extracted from brain samples/homogenates. Whole genome WNV sequences were obtained using the Illumina High Seq technology  as previously described by  and asseThe obtained West Nile virus sequence was compared with West Nile virus sequences obtained from the GenBank (NCBI) database. The analysis was conducted based on nucleotide sequences of the whole genome sequences using the maximum likelihood method and the Tamura–Nei substitution model in Mega 11 software with 1000 bootstrap replications. The evolutionary distances were computed using the maximum composite likelihood method  and are Anseriformes (Cygnus olor and Anas platyrhynchos), Passeriformes , Strigiformes , Falconiformes , Ciconiiformes (Ciconia Ciconia), and Accipitriformes (Buteo buteo). The largest group (n = 49) was constituted of birds from Passeriformes (Corvus cornix), while the least frequently diagnosed was from Corvus frugilegus (n = 2). Detailed information concerning bird species is indicated in flaviviruses.The Polish event-based surveillance system reported massive mortality  of wild birds, in September–October 2022, belonging to the following avian orders/species: Corvus cornix) collected in September 2022 were positive for WNV flavivirus from the JE serocomplex. The samples with positive results were confirmed by EURL ANSES using  RT-PCR, and the cycle threshold (Ct) values of the 17 brain samples ranged from 15.38 to 27.237 (mean 20.12). Seventeen of them were found to be infected with WNV.Samples were analyzed by RT-PCR to detect and identify RNA sequences characteristic of the West Nile virus by the method obtained from Niczyporuk, 2011. Seventeen hooded crows  were used in accordance with the protocols conducted by EURL ANSES using BWA (version 0.7.8). The sequences of the same strain were obtained in all 17 samples; therefore, the entire sequence was submitted to the NCBI GenBank database under the accession number OP804520.Corvus cornix av1-17/22, A phylogenetic tree was constructed by aligning the nucleotide sequences of the amplified fragments from the WNV genome. All ambiguous positions were removed for each sequence pair (pairwise deletion option). There were a total of 11,017 positions in the final dataset. The phylogenetic tree demonstrated WNV lineage 2 detected in Poland for the first time (GenBank accession No. OP804520 Corvus cornix) in Poland, indicating the emergence of the virus in Mazowieckie Voivodeship and the Central Mazowieckie Lowlands, on the Vistula River. The viral strain sequenced in each location is genetically similar to WNV lineage 2 strains isolated in central Europe, in particular in Hungary in 2018. Our findings indicate that the Polish strain is pathogenic in birds even if WNV infections in wild avifauna are difficult to quantify and detect by WNV-specific surveillance programs.WNV infections were pointed out for the first time in wild birds (Fringilla coelebs), 1 horse, and 14 humans [Accipiter gentilis) suspected for WNV infection (data not published). WNV circulation expansion in central, western, and northern Europe is expected to continue in the near future thanks to climatic and environmental conditions threatening Europe [From the southern mountain ranges to the Baltic Sea coast, Poland contains some of the most diverse ecosystems in Europe, with a corresponding wealth of flora and fauna. The extraordinarily intricate topography of Poland provides a healthy habitat for resident and wild avifauna. The Poland bird checklist indexes 467 bird species belonging to 27 families wintering in Poland on 31 December 2022 . Birds belonging to Corvidae are described as a reservoir for WNV in Europe ,30. This4 humans ,20, as wCorvus cornix in Poland. This analysis is extremely important information for further epidemiological analyses of wild birds in the region. The spread of the virus is dependent on ornithophilic mosquitoes (the main virus vector) and wild bird species (the reservoir of the virus). These are the links in the chain of the epidemiology of this virus. Molecular analysis of the 2022 WNV outbreak in Poland indicated 17 positive WNV fatalities in Corvus cornix, confirming WNV lineage 2 infection in Poland for the first time. From an epidemiological point of view, wild birds represent a source of multiple pathogens and various infectious diseases. In this study, the infected birds belonged to resident species. Most important from an epidemiological point of view were the birds belonging to Passeriformes. They are the primary birds involved as a reservoir of the virus and are also sensitive to WNV infection [The current study indicates the presence/circulation of WNV strains in nfection .A very large role in the spread of the virus in the world is attributed to the local movement of sedentary bird species and the long journeys of migrating birds, which cover tens of thousands of kilometers between breeding and wintering grounds ,12. AntiPasseriformes (Corvus cornix), the most common positive species in the studies, has also been indicated in 2022 in Israel [The confirmation of WNV infection in n Israel  and Italn Israel .Turdus merula), northern goshawks (Accipiter gentilis), and great grey owls (Strix nebulosa) [Corvus cornix species in September 2022 were classified as lineage 2 and cluster 2 in eastern Europe. The sequences obtained in September 2022 in Poland were closely related to several WNV strain sequences reported previously in outbreaks in 2022 in Europe: an outbreak in Italy/Piemonte and Italy/Sicily, lineage 2; a strain sequence isolated from Greece (OQ053537 and MW751840) and Serbia/Nowi Sad, 2022; and a strain sequence from goshawk isolated in Serbia in 2012 (KC407673) and Hungary in 2006 (DQ116961). We can see that this strain is being introduced globally ) and that this epidemiology is developing.In Europe, WNV in wild resident and migratory birds in Serbia with 8% of confirmed cases with specific WNV antibody detection in four mute swans, two white-tailed eagles, and one common pheasant  suggestsebulosa) . Currentebulosa) ,20. The ebulosa) . Along webulosa) . The phyPica pica) in Greece [Accipiter gentilis), white-tailed-eagles , legged gulls (larus michahellis), hooded crows (Corvus cornix), bearded parrotbills (Panurus biarmicus), and common pheasants (Phasianus colchicus) [Gyps fulvus) and little owls (Athene noctua) in Spain [This contagious virus is very dangerous for birds, and especially for geese as was confirmed and presented in 1997 in Israel, with morbidity ranging from 20% to 60%. Circulation of WNV in Hungary has been known since 1969  when then Greece . Infectelchicus) , and inflchicus)  and in gin Spain .The understanding of all factors, namely the epidemiological, ecological, and climatic conditions  together with evolutionary processes and spreading of the arboviral disease, is extremely critical and can lead us to better understand and to develop surveillance strategies to control the virus.Corvus cornix was in Poland. The Corvidae family belongs to the group of bird species most susceptible to infection with the West Nile virus and, at the same time, is considered to be the most important reservoir of the virus with high levels of viremia and massive shedding of virus particles through cloacal and oral fluids. Positive results in these species suggest that the virus may become localized because the population of these birds is significant and the risk of WNV transmission has increased. The EURL for WNV  confirmed the diagnosis. Phylogenetic analysis of the two WNV RNA genomes demonstrated that they were identical and arose as the same introduction event. WNV isolated from Poland was closely related to the WNV/Hungarian strain, OK239663 lineage 2, with 99.8% identity  during the 2022 transmission season. This study presents evidence for the presence of WNV infection in the northeastern wild bird population in Poland. The presence of genetic material of WNV demonstrates that this virus can circulate in this particular study area among wild birds in the 2023 season.Poland faced its first outbreak of WNV infection in wild avifauna (We provided the first sequence of the WNV strain circulating in the country. It belongs to the European WNV lineage 2 (GenBank accession number: PL-0P804520). Lineages 1 and 2 are the most important from a zoonotic side of infection and are isolated from many species, e.g., birds, humans, and horses.This emphasizes the need to implement a surveillance program in order to identify the other components of WNV circulation, especially the equine and human populations, to protect the elderly in particular. The objective is to deploy an early warning system ahead of a possible introduction of WNV into new geographic areas in Poland.


Generic antidepressants offer significant prescription drug cost savings compared with brand-name antidepressants, but critics of managed care interventions promoting generic medication use suggest that some generic antidepressants are not as safe or effective as the brand alternatives.To assess (a) rates of discontinuation of the initially dispensed medication and (b) disease-specific and total health care costs and pharmacy costs, comparing patients who initiated therapy with brand versus generic selective serotonin reuptake inhibitors (SSRI) or selective norepinephrine reuptake inhibitors (SNRI).Antidepressant users aged 18 to 64 years with no pharmacy claims for an SSRI/SNRI in the 180 days prior to the start of SSRI/SNRI therapy (baseline) were identified in the MarketScan database between July 1, 2005, and June 30, 2007, and were followed for 180 days (followup). All study patients met the following criteria: (a) continuously eligible from baseline through follow-up; (b) at least 1 medical claim with a primary or secondary diagnosis of major depressive disorder (ICD-9-CM codes 296.2 or 296.3) in either the baseline or follow-up period; and (c) no pharmacy claims for antipsychotic medications in the baseline period. For brand versus generic antidepressant initiators, logistic regression was used to determine the odds of 6-month therapy discontinuation, defined as no medication refills or absence of a refill for the initially dispensed medication within 1.5 times the days supply dispensed, adjusted for important covariates. Costs were measured as total plan allowed charges including member cost share. Adjusted mean  all-cause medical costs, disease-specific (claims with a ICD-9-CM diagnosis code for major depressive disorder in the primary or secondary diagnosis field) medical costs, all-cause pharmacy costs, and SSRI/SNRI antidepressant costs were compared for brand versus generic initiators using generalized linear regression models, also adjusted for baseline covariates.Of 16,659 new SSRI/SNRI users, 47.8%  initiated a brand-name medication and 52.2%  initiated a generic product. Of the 7,955 who initiated a brand-name antidepressant, 46.8%  discontinued the initially dispensed drug within 180 days, compared with 44.2%  of the 8,704 who initiated a generic. The adjusted odds of discontinuation among generic and brand drug users did not significantly differ . Adjusted all-cause 6-month average health care costs in patients initiating therapy on a generic antidepressant were $3,660  compared with $4,587  for patients initiating on a brand-name antidepressant. Adjusted average 6-month SSRI/SNRI antidepressant costs were 43.7% lower in patients initiating on a generic drug ($174 vs. $309).The likelihood of discontinuation was similar for patients who initiated therapy with brand or generic antidepressants, and short term health care costs and pharmacy costs were lower in patients starting a generic SSRI/SNRI. The results suggest that the use of generic antidepressants as first-line agents in the treatment of major depressive disorder is associated with continuation rates similar to initiation with brand antidepressants but with lower health care costs.