The HIV pandemic impacts the lives of millions and despite the global coordinated response, innovative actions are still needed to end it. A major challenge is the added burden of coinfections such as viral hepatitis, tuberculosis and various sexually transmitted infections in terms of prevention, treatment and increased morbidity in individuals with HIV infection. A need for combination prevention strategies, tailored to high-risk key populations arises and technology-based interventions can be a valuable asset. The COVID-19 pandemic challenged the delivery of existing services and added stress to existing public health and clinical structures but also highlighted the potential of exploiting technical solutions for interventions regarding infectious diseases. In this paper we report the design process, results and evaluation findings from the pilots of ‘RiskRadar’—a web and mobile application aiming to support combination prevention, testing and linkage to care for HIV, viral hepatitis, various sexually transmitted infections and tuberculosis.RiskRadar was developed for the INTEGRATE Joint Action’s aim to improve, adapt and pilot innovative digital tools for combination prevention. RiskRadar was designed iteratively using informed end-user-oriented approaches. Emphasis was placed on the Risk Calculator that enables users to assess their risk of exposure to one or more of the four disease areas, make informed decisions to seek testing or care and adjust their behaviours ultimately aiming to harm/risk reduction. RiskRadar has been piloted in three countries, namely Croatia, Italy and Lithuania.RiskRadar has been used 1347 times across all platforms so far. More than 90% of users have found RiskRadar useful and would use it again, especially the Risk Calculator component. Almost 49.25% are men and 29.85% are in the age group of 25–34. The application has scored 5.2/7 in the User Experience Questionnaire, where it is mainly described as “supportive” and “easy-to-use”. The qualitative evaluation of RiskRadar also yielded positive feedback.Pilot results demonstrate above average satisfaction with RiskRadar and high user-reported usability scores, supporting the idea that technical interventions could significantly support combination prevention actions on Sexually Transmitted Infections. HIV, viral hepatitis, Tuberculosis (TB) and other Sexually Transmitted Infections (STIs) continue to pose major public health challenges globally and gaps in the coverage of the continuum of care services are widely acknowledged , 2. The However, one-size-fits-all approaches have failed to support the broad and complex spectrum of HIV, viral hepatitis and STIs prevention needs ; EffectiThe INTEGRATE Joint Action (JA)  has implThe RiskRadar application design process see Fig.  began wiThese shortlisted tools drove the initial proposal of the RiskRadar structure; the adaptation and integration process aimed to accommodate all four disease areas and the pilot-specific target audiences, while considering the pre-existing tools’ strengths, weaknesses and perceived effectiveness to address the different information and prevention aspects. The overall technical design of RiskRadar was based on informed User eXperience (UX), end-user oriented design and communication approaches –22. The Factsheets (FS) offering basic information about the four disease areas , (b) Risk Calculator (RC) that helps users assess their risk of exposure to the different diseases according to their behaviour and to make informed decisions in order to adjust their protective behaviours, (c) PrEP and Undetectable = Untransmittable (U=U) information, (d) Test Finder (TF) with filtering capabilities per disease and country, (e) Partner Notification (PN), an anonymous, non-traceable SMS service to notify one’s partners that can be accessed via a unique random code  that only diagnosed patients receive from an authorized healthcare professional, (f) Reminders (only featured in the mobile app) for the users with an estimated risky behaviour to get tested every 6 months that can be disabled at any time, and (g) Evaluation Questionnaire (EQ) [The RiskRadar components can be summarized as following: (a) ire (EQ)  to colleThe functionality of the Risk Calculator component aimed to facilitate the individual risk assessment based on the decision trees (a) described in WHO and ECDC guidelines –26 and  and on Italy’s and Lithuania’s Ministry of Health websites. RiskRadar has been presented in multiple occasions at national and regional meetings to public health specialists and other involved stakeholders. Targeted promotion was handled by each organisation as needed, using available communication channels and through their day-to-day interactions with potential end-users. To this end, paid campaigns were launched on Grindr, a prominent social networking app for gay, bi, trans and queer people in Italy, and on the gay.it website. Moreover, the toolkit was presented to people accessing low-threshold service locations where it could gain visibility among vulnerable risk groups.RiskRadar was officially launched on July 6th, 2020 -after some months’ delay due to the COVID19 pandemic; the piloting phase lasted until December 31st, 2020. In total, it was accessed 817 times via web, while 270 and 260 users have downloaded the app on Android and iOS respectively. The Risk Calculator component  questions about its perceived usefulness and (b) a short user experience evaluation questionnaire (UEQ) . This coRegarding the qualitative evaluation, in Italy 10 participants were interviewed online by Italian pilot partners; 80.00% were men  and 40.00% of them belonged in the 45–54 age range followed by the 35–44 and over 75 age range (20.00% each). Self-reported computer skills were average and above. Figure In Lithuania, face to face focus groups were organised instead, involving 12 patients with diagnosed addictive disorders, however only 7 of them completed the UEQ after the meetings. All respondents were males; 71.40% belonged in the 35–44 age range and 28.60% in the 45–54 range. 6 participants self-assessed as having “bad” or “moderate” computer skills. Overall, the tool received positive feedback . In the future, a controlled study engaging a representative sample of the targeted populations could provide more robust results. Furthermore, low computer literacy, prevalent in some vulnerable groups, has been identified as key factor which might hinder the uptake of similar interventions. Moreover, it should be highlighted that as COVID-19 pandemic affects most key populations disproportionately by hindering or at least delaying access to testing and treatment, especially for vulnerable populations , 32 and As a whole, we consider RiskRadar’s uptake to be positive, taking into account the difficulties introduced by the COVID-19 pandemic, especially regarding the dissemination of the tool among the target groups. The majority of users in the pilots originate from Italy, partly due to the timely and coordinated efforts of the partners and the experience and traction gained from previous similar campaigns, but also because of the disparities in population size. All pilot countries managed to intensify their efforts during the last months while targeting key population groups and supporting their use of RiskRadar, which was clearly shown in the daily breakdown of usage metrics.The overall evaluation of the tool was satisfactory with RiskRadar’s understandability and user friendliness scores being rated as highly positive. Furthermore, INTEGRATE’s Scientific Committee and Advisory Board members have suggested the additional expansion of the RiskRadar components to other combination prevention issues such as HIV and STIs self-tests, chemsex, harm reduction advice and services etc.Outlining future work paths, RiskRadar’s further development should focus on (a) following an adaptive model to address the variety of needs per population group, (b) systematic evaluation and (c) long-term sustainability plans to maximise its impact.RiskRadar targets population groups at risk for HIV, hepatitis, STIs and TB with clear and simple combination prevention messages through several modular core components. Based on the overall experience of the RiskRadar development, we argue that RiskRadar and similar interventions have the potential to provide support towards combination prevention efforts, provided that considerable resources to ensure its accuracy, reliability, relevance and usefulness are invested in the future. As the COVID-19 pandemic has also clearly highlighted, the need for timely monitoring, prevention, testing and risk assessment constitute a necessity that ICT technologies can significantly address.


Novel Severe Acute Respiratory Syndrome-Corona Virus-2 infection (SARS-CoV-2) is an acute respiratory and infectious disease. This perspective aims to provide a basic understanding of the inflammation caused by SARS-CoV-2 and its relation to the trigeminal ganglion (TG). The virus enters through the mucous membranes of the orofacial region and reaches the TG, where it resides and takes control of its peptides including Substance P (SP). SP is the main neuropeptide, neuromodulator, and neuro-hormone of TG, associated with nociception and inflammation under noxious stimulus. SP release is triggered and, consequently, affects the immune cells and blood vessels to release the mediators for inflammation. Hence, cytokine storm is initiated and causes respiratory distress, bronchoconstriction, and death in complicated cases. Neurokinin-1 Receptor (NK-1R) is the receptor for SP and its antagonists, along with glucocorticoids, may be used to alleviate the symptoms and treat this infection by blocking this nociceptive pathway. SP seems to be the main culprit involved in the triggering of inflammatory pathways in SARS-CoV-2 infection. It may have a direct association with cardio-respiratory rhythm, sleep-wake cycle, nociception, and ventilatory responses and regulates many important physiological and pathological functions. Its over-secretion should be blocked by NK-1R antagonist. However, experimental work leading to clinical trials are mandatory for further confirmation. Here, it is further proposed that there is a possibility of latency in SARS-CoV-2 virus infection if it is acting through TG, which is the main site for other viruses that become latent. It provides somatosensory innervation to the orofacial region. So any alteration in its secretion in response to viral infection may result in symptoms in the orofacial region (Severe Acute Respiratory Syndrome-Corona Virus-2 (SARS-CoV-2) is responsible for causing Corona virus infection COVID-19) which was first reported in Wuhan, China in December 2019 but went on to affect the whole world 9 which w, 4. Compl region   gene (SP is a “brain-gut” hormone and the first inflammatory neuropeptide discovered in 1931 by Euler and Gaddum . It belo-1) gene . SP is r-1) gene , 16, 17.-1) gene . SP alonNeurokinin-1 Receptor (NK-1R) is the receptor for SP and most of the functions of SP are elicited only after binding with its receptor. Both SP and NK-1R are abundant in the central nervous system (CNS), peripheral nervous system (PNS), and enteric system . It alsoAny noxious stimulus from the orofacial structures, such as the eye, nose, or mouth, is mainly transmitted by the TrN . TG neurvia afferent fibers to the trigeminal spinal caudalis (Vc) nucleus of the brainstem. Here, they synapse with the second order neurons that project to the somatosensory and limbic cortices via the thalamus. Inflammation of orofacial tissues that are innervated by the TrN can modify the activity of trigeminal afferent neurons, consequently causing ectopic firing and raised sensitivity of noxious stimuli. Sensitization is facilitated by many mediators such as neurotrophic factors or neuropeptides at nerve endings, such as SP, CGRP, and serotonin  infects the ocular mucosal membrane and orofacial region and continues to infect the sensory neurons leading to a latent infection in TG . The virLatent HSV-1 DNA has been detected in the CNS of human postmortem, and infection with HSV has been correlated with the development of neurodegenerative diseases. However, whether HSV can directly reactivate in the CNS and/or infectious virus can be transported to the CNS following reactivation in peripheral ganglia has been unclear. Viral proteins were detected in neurons of the TG. These results suggest that infectious virus is transported from the TG to the CNS following reactivation but does not exclude the potential for direct reactivation in the CNS .SP is the first to react in response to a noxious stimulus. It is a rapid and immediate defense in the survival system. In experimental studies, NK-1R deficient mice exhibited reduced pulmonary inflammation as compared to the controls . Immune SP/NK-1R may control the breathing activity in neonates, evident from the raised immunohistochemical SP expression in the brainstem tissues after postmortem studies in control infants as compared to Sudden Infant Death syndrome (SIDS) victims in my previous study at Centro Lino Rossi, University of Milan, Italy , 16. SP Reactivation occurs in Herpes and HIV but is not common in other types of viruses. It has not been speculated to exist in SARS-CoV-2, at least in a shorter duration of time. But if it proves to be the case then it may cause infection again at a later stage in life. Upon contracting the virus in orofacial parts, the virus reaches the TG through TrN and may reside there, taking control of the release of its peptides, particularly SP. The vaccines may also not be of much help in eradication of the disease . AntibodAntibody therapies such as passive immunization and Intravenous immunoglobulin therapy may also not be useful in these circumstances. In that case, the promising strategy would be the use of antiviral drugs, corticosteroids, and SP/NK-1 blockers to cease the inflammatory responses and attack the latent viruses as well. Although this possibility has limited chances, it cannot be ruled out totally due to its entry route through trigeminal nerves and ability to target TG like other latent viruses such as Herpes Simplex Viruses. We have to keep an eye on this phenomenon as well. Whether the virus becomes latent upon reaching TG or not is one aspect that needs to be explored, but there are more possibilities that this virus reaches TG, modulates SP release, and initiates inflammatory mechanisms.Another compelling argument is that these viruses are highly mutating and transforming, making them a difficult target for vaccines. Vaccines are successful for those viruses that mutate less, have strictly humans as host, and have only one stable antigenic type. Common cold viruses, including SARS-CoV-2, have at least 100 antigenic types and a high mutation rate.Serotonin and CGRP also co-exist with SP and should be explored as well. The abnormal release of SP from CN V may be associated with the acute respiratory symptoms in SARS-CoV-2 infection. SP may trigger the immune cells to release the inflammatory mediators in CNS as well as in the respiratory system, causing cytokine storming, inflammation, lung injury, and broncho-constriction. In complicated cases it may lead to cardiac failure. TG stimulation by virus may also increase the conductance of sodium ions and decrease the conductance of potassium ions in the cells leading to hypernatremia, hypokalemia, and dehydration. This electrolyte and fluid balance will exert pressure on the kidneys and pumping of the heart, causing failure and death. Maintaining electrolyte and fluid balance is necessary to manage these patients. Potassium and magnesium should be added in their dosage. High sunlight exposure and stress may also reactivate the virus. But all these phenomenon need to be evaluated and clinical trials are required.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.RM has conceived the idea and proposed the main theory. MK, AB, NA, HN, LM, TA, AR, and RJ have critically evaluated, contributed in the write up and finalization of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Astroviruses (AstVs) are non-enveloped, positive single-stranded RNA viruses that cause a wide range of inflammatory diseases in mammalian and avian hosts. The T = 3 viral capsid is unique in its ability to infect host cells in a process driven by host proteases. Intercellular protease cleavages allow for viral egress from a cell, while extracellular cleavages allow for the virus to enter a new host cell to initiate infection. High-resolution models of the capsid core indicate a large, exposed region enriched with protease cleavage sites. The virus spike protein allows for binding to target cells and is the major target for naturally occurring and engineered neutralizing antibodies. During maturation, the capsid goes through significant structural changes including the loss of many surface spikes. The capsid interacts with host membranes during the virus life cycle at multiple stages such as assembly, host cell entry and exit. This review will cover recent findings and insights related to the structure of the capsid and its function. Further understanding of the viral capsid structure and maturation process can contribute to new vaccines, gastric therapeutics, and viral engineering applications. Astroviridae consists of two genera, Mamastrovirus and Avastrovirus, that infect a broad range of mammalian and avian hosts, respectively . T. T29]. Ttrovirus . This retrovirus ,32. TAsttrovirus . This inerotypes . More inWhile there have been no new astrovirus structures solved using X-ray crystallography in the last four years, there have been several recent studies using existing structural information to model the evolution of AstV structures ,35. A laWhen the human astrovirus capsid assembles, it is made up of 180 subunits and maintains its T = 3 architecture throughout its lifecycle. The VP90 state of the virus includes all five domains of the virus capsid protein (CP), with the C-terminal acidic domain somewhere at the exterior of the virus, allowing it be accessible to cellular membranes and caspases. The VP90 state of the virus has been difficult to isolate, but several studies have used recombinant systems to attempt to isolate this state of the capsid. A vaccinia virus system was used to express the HAstV-2 ORF2 sequence in BSC-40 and LLCMK2 cells, which showed the generation of aggregations of virus-like particles (VLPs) in the infected cells . There hAfter maturation, VP70 state of the virus can egress from the cell. This has allowed for the isolation of the HAstV-8 VP70 particle from protease-inhibitor-treated cell culture lines and structural analysis by cryo-EM . Due to Post-maturation structural analysis indicated that the roughly 30 amino acid regions that present at the N-terminus of VP27 but are absent from VP25 are necessary to anchor the spike to the core domain, while VP25 on its own has no covalent or noncovalent interaction with the capsid core. This means that the spikes present in mature capsid are almost entirely VP27 . It is uOne of the outstanding questions regarding the AstV capsid is its ability to bind to host cell receptors to initiate viral entry. Along with protecting the viral genome, the AstV capsid must be able to bind with the host cell to initiate the entry process. Several earlier studies have looked into candidate receptors, with the spike domain having the properties to mark it as the candidate binding domain . The strAs discussed above, membranes likely play an important role in AstV capsid assembly, but there is also evidence for capsid–membrane interactions that occur during viral entry and exit of an infected cell. In terms of viral entry, many non-enveloped viruses use a membrane-interacting domain or protein complex to allow for the delivery of the viral genome across membrane, such as other gastroenteric pathogens with the rotavirus VP4 being a prime example . One of Mammastrovirus strains. The sequence ranges in length from 64 amino acids in murine AstVs to 148 amino acids in bat AstVs, with large variabilities in the chemical properties of the amino acids that make up the sequence. Some ORFX products are expected to have transmembrane regions in the C-terminus, while others have no predicted activity, indicating that membrane integration may not be necessary for the function. Structural studies on this protein product across the AstV family would be highly valuable to further understand the functions and underlying mechanisms of this novel ORFX product.AstV proteins may also play a role in the ability for the virus to exit the cell. While there is evidence the transition from the VP90 to VP70 state of the capsid allows for the virus to exit the cell in a non-lytic manner, other structural/functional relationships related to this transition are unknown . A seconSerological analysis in pediatric populations across the United States indicate a prevalence of neutralizing antibodies ranging from 42% of the surveyed population to HAstV-3 to 94% to HAstV-1 ,47. ThisAs our understanding of AstV structural proteins expand, there is great potential for developing applied technologies using these proteins. One technology that can directly benefit from this information is the development of new astrovirus vaccines for either mammalian or avian hosts ,53,54,55Recent developments in AstV reverse genetics systems allow for precision engineering and good recovery from expression cell lines ,57. EvenThe maturation process of the AstV capsid provides an interesting direction for protein and viral vector design. Rather than being mediated by viral proteases, the astrovirus capsid changes its structure and property throughout the virus life cycle in a process driven by the host environment. Other virus systems have been engineered to utilize host proteases in order to function, such as adeno-associated virus (AAV) derived genetic therapy vectors that only transduce cells when activated by tumor environments through metalloproteinases ,60. The Since the first discovery of human astroviruses in 1975, our understanding of the astrovirus capsid structures and functions has continued to expand through the years, especially in areas related to laying down the structural framework of the capsid protein and elucidating maturation-related structural changes as well as mechanisms of antibody neutralization. Exciting new insights have begun to emerge regarding astrovirus receptor binding, host entry, virus assembly and cellular exit, although a detailed mechanistic understanding is still lacking. Compared to other gastroenteric viral pathogens, HAstVs are certainly under-characterized at this time. Some of the most urgent questions involving the capsid that remain to be answered are, but not limited to: (1) the underlying mechanism that allows mature astrovirus to gain infectivity after extracellular protease processing of the viral capsid; (2) the identity of the cellular receptor for the virus and its interaction with the capsid; (3) the mechanism of astrovirus internalization and host membrane penetration mediated by the viral capsid and/or associated factors; (4) the interaction of VP90 with the viral genome and other viral/host factors that leads to the assembly of a precise T = 3 particle; (5) the structure and function of the acidic domain of the astrovirus CP; and (6) the mechanism of intracellular trafficking and non-lytic release of the astrovirus particle. Considering recent advances in cellular imaging, electron tomography, molecular labeling, mass spectrometry, genetics, and other techniques in structural and molecular biology, we are optimistic that accelerated progress will be made in various fronts. The discovery of neuropathogenic astroviruses such as MLB and VA will also likely catalyze more intensive research efforts in studying the fundamental virus infection mechanisms.


To initiate infection, a virus enters a host cell typically via receptor-dependent endocytosis. It then penetrates a subcellular membrane, reaching a destination that supports transcription, translation, and replication of the viral genome. These steps lead to assembly and morphogenesis of the new viral progeny. The mature virus finally exits the host cell to begin the next infection cycle. Strikingly, viruses hijack host molecular chaperones to accomplish these distinct entry steps. Here we highlight how DNA viruses, including polyomavirus and the human papillomavirus, exploit soluble and membrane-associated chaperones to enter a cell, penetrating and escaping an intracellular membrane en route for infection. We also describe the mechanism by which RNA viruses—including flavivirus and coronavirus—co-opt cytosolic and organelle-selective chaperones to promote viral endocytosis, protein biosynthesis, replication, and assembly. These examples underscore the importance of host chaperones during virus infection, potentially revealing novel antiviral strategies to combat virus-induced diseases. To cause infection, an incoming viral particle engages a receptor that is expressed on the plasma membrane of the host cell . This inDespite these differences, one emerging common principle is that viruses co-opt host molecular chaperones to promote distinct entry steps. Molecular chaperones typically function to fold or unfold a protein, helping to achieve its native conformational state or to untangle its structural configuration . MoleculPolyomaviruses (PyVs) cause devastating human diseases, particularly in immunocompromised individuals . Well-stSV40 is composed of 72 pentamers of the VP1 coat protein that encloses its 5 kilobase-pair double-stranded DNA genome ,7; each Recent studies on the mechanism of the SV40 ER escape and ensuing cytosol-dependent disassembly have revealed a network of ER-associated and cytosolic chaperones as responsible for these processes. Specifically, upon reaching the ER lumen from the cell surface, ER luminal redox factors belonging to the protein disulfide isomerase (PDI) family reduce and isomerize the disulfide bonds of the viral particle B, step 1Once SV40 has integrated into the ER membrane, it reorganizes the ER to form an exit site  from which it gains access to the cytosol ,22,23,24In parallel, SV40 induces several DNA J protein family members  to accrue in the foci ,23. A J Upon reaching the cytosol, the SV40 capsid proteins are partially disassembled by the cytosolic dynein motor adaptor called BICD, which surrounds the ER-foci B, step 5In addition to PyV, the human papillomavirus (HPV) is yet another DNA tumor virus that co-opts host chaperones during infectious entry. HPV is a highly prevalent virus, infecting approximately 80 million adults in the United States . It beloStructurally, HPV is a nonenveloped double-stranded DNA tumor virus composed of 72 pentamers of the major capsid protein L1. The L1 pentamers encase up to 72 copies of the minor capsid protein L2. Together, these encapsulate the 8 kilobase-pair double-stranded DNA genome . The assAfter delivery to the endosome, the viral capsid proteins begin to dissociate from the viral genome in a pH-dependent manner. This disassembly is partial, as L2 and a subset of L1 remains associated with the viral genome ,47. PostThe action of host chaperones regulates the earliest stage of HPV entry. Specifically, conformational changes of the viral capsid at the plasma membrane are mediated in part by the peptidyl-prolyl isomerase activity of cyclophilin (CyP), specifically cyclophilin B . These cAfter the HPV particle is partially disassembled in the endosome, L2 is inserted into the endosomal membrane resulting in exposure of L2 to the cytosol. This is a decisive step because the cytosol-exposed L2 engages the host sorting factors which target the virus along a productive infection pathway. Topologically, when L2 is inserted into the endosomal membrane, its N-terminus remains associated with the viral genome while the C-terminus is exposed to the cytosol . StrikinThe delivery of HPV to γ-secretase in the endosome relies on the cytosolic γ-secretase adaptor p120 catenin, which normally targets cellular transmembrane proteins to γ-secretase ,62,63. IIn addition to the double-stranded DNA tumor viruses PyV and HPV, single-stranded RNA viruses—including flaviviruses and coronaviruses—also exploit the intrinsic activities of host molecular chaperones to support infection. Within the flavivirus family, its prominent members include Dengue virus (DENV), Zika virus (ZIKV), Japanese encephalitis virus (JEV), West Nile virus (WNV), yellow fever virus (YFV), and tick-borne encephalitis virus (TBEV). These viruses are among the most crucial arthropod-borne viruses that cause a myriad of human diseases . In the case of DENV, this flavivirus initiates infection by interacting with a receptor on the cell surface that leads to internalization , step 1 Once the replication compartment is formed and replication of the viral RNA genome is completed, the new viral progeny begins to assemble , step 5 The actions of molecular chaperones have been reported to promote many of these distinct DENV entry steps. For instance, cytosolic Hsp70 chaperone, in concert with dedicated DNA J proteins, is thought to facilitate viral endocytosis, protein biosynthesis, and assembly . MechaniAlong with cytosolic chaperones, ER-associated molecular chaperones also play critical functions during flavivirus infection. For instance, the ER-resident EMC transmembrane chaperone contributes to DENV and ZIKV infection. EMC does so by promoting the biogenesis of DENV and ZIKV multipass membrane proteins NS4A and NS4B , step 3 Coronaviridae which contains four distinct subgroups: alpha, beta, gamma, and delta CoV, with the alpha and beta subgroups being the most prevalent. CoV infections in humans typically lead to mild symptoms such as the common cold, including infections caused by human coronavirus OC43 (HCoV-OC43), human coronavirus HKU1 (HCoV-HKU1), and human coronavirus 229E5 (HCoV-229E5) [Coronaviruses (CoVs) are enveloped, positive-sense, single-stranded RNA viruses. They are part of the subfamily V-229E5) . HoweverV-229E5) . AdditioV-229E5) . Most reV-229E5) . Thus, tStructurally, CoVs are approximately 100 nm in diameter with 5′-capped genomes that range between 26 to 32 kb—the largest of all RNA virus genomes . PredictUpon release into the cytosol, the ER then assumes a key role during the replication cycle of CoVs. The viral RNA genome engages the ER membrane, where it is thought to undergo cotranslational translocation , step 3.The subgenomic RNAs encoding S, M, and E are ejected from the DMV via an NSP3-containing pore that spans the double-membrane of the DMV , step 6,Given that the ER executes a key function in CoV infection, it is perhaps not surprising that ER-associated chaperones promote distinct viral entry steps. In fact, ER chaperones have been reported to facilitate the earliest step of viral entry. For instance, using gain- and loss-of-function approaches, the spike protein of both MERS-CoV and the bCoV-HKU9 beta coronavirus was found to engage BiP at the cell surface , step 1 At the ER, CoVs commandeer the activities of ER-associated chaperones, which are generated through virally-induced ER stress. SARS-CoV, for example, triggers the unfolded protein response (UPR) via its S protein ,108. TheThe importance of ER-associated chaperones during CoV infection has been reignited by COVID-19. Indeed, the global effort to identify host proteins exploited by SARS-CoV-2 that promote infection has uncovered a myriad of ER-dependent chaperones, including EDEM3, ERLEC1, RTN4, Sigma-1 receptor, that play essential (but poorly characterized) roles during SARS-CoV-2 entry ,113. WitExploiting host chaperones is a common strategy used during the entry of many viruses. As described in this review, both DNA and RNA viruses take advantage of host chaperones in a variety of ways to ensure a productive viral life cycle. The chaperone functions hijacked by these viruses are used to support different entry steps, including cell surface attachment and endocytosis, unfolding and disassembly, as well as membrane penetration and escape of the input viral particle. Moreover, these molecular chaperones are co-opted to assist in the biogenesis and folding of new virus proteins which are essential for the formation of the new viral progenies.In many instances, a virus simply uses a host chaperone’s well-established canonical function during entry. For example, the PDI oxidoreductase reduces and isomerizes the disulfide bonds in PyVs in the ER lumen to prime the virus for ER escape—PDI deploys this same activity on nascent cellular polypeptide chains as they are undergoing redox-dependent folding. Similarly, the ER-resident BiP promotes folding of the polyproteins of SARS-CoV-2, in much the same way BiP acts as a general chaperone to fold cellular clients. Establishing that viruses exploit the canonical activity of host chaperones provides a distinct advantage in developing rational antiviral therapies—any drugs that inhibit the normal functions of these chaperones can now be repurposed and tested to assess if they can likewise block virus infection. Future investigations along this line of research may prove useful.By contrast, it is entirely possible that viruses exploit the noncanonical functions of host chaperones to support their infection. For instance, although γ-secretase is critical during HPV infection, the canonical proteolytic activity of this transmembrane protease—central to its normal cellular function—is not necessary during HPV entry. Instead, γ-secretase possesses a new “insertase” activity that promotes membrane insertion of HPV, enabling the virus to transport along a productive route . This neIn sum, these examples revealing unanticipated noncanonical functions of the host chaperones only further underscore the remarkable history of studying virus-host cell interaction. Not only do these studies provide critical insights into the basic mechanism of viral pathogenesis, they also expand the full repertoire of functions hidden in the host chaperones that might not have been revealed if not for these toxic pathogens.


The relevance of local twitch response (LTR) during dry needling technique (DNT) is controversial, and it is questioned whether LTR is necessary for successful outcomes. Furthermore, because the LTR during the deep DNT may be evoked with different intensities, it is unknown whether the magnitude of LTR intensity is associated with optimal clinical results, especially concerning to the effects of joint maximal range of motion (ROM). This study aimed to (i) determine whether visual inspections can quantify the LTR intensity during the DNT through a qualitative ultrasonography scale of LTR intensity , and (ii) assess the differences of US-LTR scale associated with changes in the maximal joint ROM.Using a cross-sectional design, seven asymptomatic individuals were treated with DNT in the latent myofascial trigger point in both medial gastrocnemius muscles. During DNT, three consecutive LTRs were collected. The US-LTR scale was used to classify the LTRs into strong, medium, and weak intensities. The categories of US-LTR were differentiated by the velocity of LTRs using the optical flow algorithm. ROM changes in ankle dorsiflexion and knee extension were assessed before and immediately after DNT.p < 0.05). A significant difference in velocity was observed between US-LTR categories (p < 0.001). A significant difference in the ROM was observed between the strong and weak-medium intensity (p < 0.05).The US-LTR scale showed the third LTR was significantly smaller than the first one (The present findings suggest that the LTR intensity can be assessed using a qualitative US-LTR scale, and the effects of DNT on joint maximal ROM is maximized with higher LTR intensity. This study reports a novel qualitative method for LTR analysis with potential applications in research and clinical settings. However, further research is needed to achieve a broader application. The Myofascial trigger points (MTrPs) have been defined as hyperirritable nodules in tense bands of skeletal muscle that exhibit motor, sensory, and autonomic components and classified as either active or latent . MTrPs hThe prevalence of latent MTrPs in the lower limb of asymptomatic people has been reported to be around 78 % , whereasActive and latent MTrPs have been widely treated by the use of dry needling techniques (DNT) to increase ROM in healthy individuals and those with neurological and musculoskeletal disease , 14, 28.To better understand the physiology of LTR, clinical tools are necessary. For that, ultrasound-guided trigger point puncture has been proposed to detect LTR, taking into account that the detection of LTR is not always evident by visual or palpation assessment . However2) were recruited from San José Hospital, Santiago Chile (employees only). Ethical authorization was obtained from the Ethics Committee of the Northern Metropolitan Health Service of Santiago, Chile. All participants agreed to participate in this study and signed an informed consent form. All methods were carried out in accordance with relevant guidelines and regulations. The study was registered in ClinicalTrials.gov (NCT02824991) and reporting adhered to the STROBE guidelines.Using a cross-sectional and non-probability sampling, seven healthy young men , with a randomized order. Participants lay in a prone position with the ankle fixed at 90º . A local examiner (CCM) ensured the maintenance of ankle and knee angle during the DNT intervention. The localization of latent MTP at the medial gastrocnemius was identified by determining a palpable and hypersensitive taut band criteria , 43. TheThe ankle dorsiflexion and knee extension maximal ROM were assessed (with a randomized order) before and immediately after DNT intervention. The same physical therapist (PB) performed all ROM measurements, who was blinded about the intensities of LTR collected during the DNT. The ankle dorsiflexion and knee extension maximal ROM of both lower limbs were assessed using weight-bearing and active knee extension test, respectively, as an estimate of medial gastrocnemius and hamstring extensibility. A universal goniometer  was used to perform the joint ROM testing. Three measurements were performed for each test, which has shown to have an excellent inter-rater reliability for the ankle and knee maximal ROM .For the active knee extension test, the participants were in a supine position with the contralateral knee extended and the hip in a neutral position and the ipsilateral hip and knee flexed to 90° . A belt For the weight-bearing ankle dorsiflexion ROM test, the participants were instructed to keep the knee of the tested leg extended and then to maximally flex their tested ankle while keeping their heel on the floor. The weight-bearing test using an standard goniometer has been reported to be a valid test to assess the ankle dorsiflexion maximal ROM , 41. A vSonographic data was processed using Matlab® scripts . The LTRs were examined offline by a physical therapist (CCM) with 5 years of experience in muscle ultrasound imaging. The Camtasia software  was used to select the three LTRs for later analysis. A visual criterion was established to differentiate muscle movement due to the DNT and LTRs, considering the vertical and horizontal movement in the deep aponeurosis. It should be noted that when the dry needle is inserted into the muscle, only a vertical displacement is produced.https://la.mathworks.com). The parameters used were a sigma value of 1 and a sample step of 1. Two regions of interest with a standard size for width  and adapted size por height depended on the thickness of the deep aponeurosis (four times of deep aponeurosis thickness), were manually selected at the center of the echogram map by including deep aponeurosis (i.e. hyperechoic line) and its surrounding muscle borders . The maximal velocity between both regions of interest was determined and considered for the statistical analysis.The deep aponeurosis was tracked by the Lucas–Kanade optical flow algorithm with affine optic flow extension . The LTRAfter deep aponeurosis motion processing, two raters with low experience (novices) in ultrasound (RNC and CT) classified LTR in three intensity categories: weak, medium, and strong. For this purpose, one rater with high experience (expert) in ultrasound (CCM) gave an explanatory session (2 hours) to the novice raters concerning the visual interpretation (in b-mode recordings) of LTR with different intensities. The instruction was focused on the velocity of the deep aponeurosis. Three LTR were presented with deep aponeurosis motion velocities of 2.8 mm/s , 3.3 mm/s , and 5.4 mm/s . The qualitative scale (US-LTR) was practiced by the novices in three videos prior to the analysis. The high and low experienced raters were blinded to the maximal ROM values after DNT, and the values of maximal velocity of LTR.n = 7 legs and strong: n = 7 legs). Unpaired testing was conducted to compare groups (week-medium vs. strong) because only three participants were members of both groups.All data were analyzed with the SPSS software . The normality of data distribution was evaluated using the Shapiro-Wilk test. To compare the intensities of the three consecutive LTRs recorded through the US-LTR scale, the Friedman test and multiple comparisons with Dunn’s correction were used. The agreement of the US-LTR scale between the expert and the two novices, and between novices was evaluated using the Kappa statistic; and the agreement result was interpreted as none (≤ 0), none or weak agreement (0.01–0.20), fair (0.21–0.40), moderate (0.41–0.60), substantial (0.61–0.80), and almost perfect (0.81–1.00) . To compp-value of < 0.05 was considered statistically significant. The data was expressed as mean and 95 % confidence interval unless stated otherwise. For parametric comparison, the effect sizes was established by calculating Cohen’s d  to indicate small, moderate, or large effects, respectively [r conversion of the z-score  to indicate small, moderate, or large effects, respectively [A ectively . For nonectively .The sample size needed for this study was calculating using GPower software, version 3.1.9.2 . Considering a previous reported large effect size on maximal ROM after treatment of latent MTrPs , seven pp = 0.008; r = 0.8, large effect), but not between the first and second , and second and third LTRs   Fig. A. The poct) Fig. A.Fig. 2p = 0.013) , but not between the first and second , and second and third LTRs  . The post-hoc analysis showed a significant difference and large effect size for all comparisons   Fig. B. The poct) Fig. B. The tr.8) Fig. C.p < 0.001)  , inter-rater agreement of US-LTR scale  between the expert and the two novices was substantial  Fig. . The agr01) Fig. .Fig. 3Ap < 0.001)  ly) Fig. B.Fig. 4The aim of this report was to determine whether the LTR intensity during DNT can be quantified through visual inspection using a qualitative ultrasonography scale of LTR intensity , as well as to assess the differences between the intensities of LTRs associated with changes in maximal joint ROM. The US-LTR scale and automatic tracking of the deep aponeurosis (gold standard) showed a significant difference in LTR intensity between the first and third LTR. Moreover, the findings of this study show good agreement with the qualitative US-LTR scale between the expert in ultrasound and novices. Furthermore, this study indicated that the intensities of LTRs  may involve different magnitudes of maximal ROM changes after DNT interventions. The present result supports the hypothesis that the US-LTR scale is a valid tool to quantify the intensity of LTR, and a greater LTR intensity implies a greater maximal ROM after DNT.The US-LTR scale was sensitive enough to identify a decrease in intensity between the first and third LTR. The decrease in LTR intensities observed in this study agrees with a recent study where the LTR intensities were assessed with surface electromyography during the application of DNT over latent MTrPs in the medial gastrocnemius . The decin vivo models [We observed a significant difference in the maximal joint ROM between the strong and weak-medium intensities. Improved joint flexibility following DNT has been previously reported , 2, showo models , 44. TheRegarding the clinical implications of these results, ultrasound imaging technology is currently increasing in use among health professionals to assess muscle function and puncture guidance , 39. BecThis study has several limitations. First, the joint torque and electromyography were not assessed. Therefore, it is unclear the time related of LTR with proprioceptive afferents reflex and the motor unit activity of the investigated muscles. Second, the effect of DNT on maximal ROM was not counteracted with a placebo group  , which sFinally, future randomized clinical trials should investigate the clinical implications of LTR intensities during the dry-needling intervention in musculoskeletal and neurological diseases.The present findings suggest that the LTR intensity can be assessed using a qualitative US-LTR scale, and a greater LTR intensity implies a greater ROM post dry needling intervention. This study reports a novel qualitative method for LTR analysis with potential applications in research and clinical settings. However, further research is needed to achieve a broader application.


The majority of plant viruses are transmitted by hemipteran insects. Bacterial symbionts in hemipteran hosts have a significant impact on the host life, physiology and ecology. Recently, the involvement of bacterial symbionts in hemipteran vector-virus and vector-plant interactions has been documented. Thus, the exploitation and manipulation of bacterial symbionts have great potential for plant viral disease control. Herein, we review the studies performed on the impact of symbiotic bacteria on plant virus transmission, including insect-bacterial symbiont associations, the role of these bacterial symbionts in viral acquisition, stability and release during viral circulation in insect bodies, and in viral vertical transmission. Besides, we prospect further studies aimed to understand tripartite interactions of the virus-symbiotic microorganisms-insect vector. This is based on the length of the period the vector can harbor infectious particles, which ranges from minutes to hours (non-persistent), days (semi-persistent) as well as for a lifetime and even inheritance by insect progeny (persistent). For non-persistent and semi-persistent viruses, viral particles are mainly retained by the vector in the stylet  or foregut, respectively , blood-feeding tsetse flies, cockroaches, weevils, and certain generas of ants . Therefot groups . In addit groups , or by bt groups . Consequt groups . With ret groups . Besideschanisms .Buchnera aphidicola (hereafter referred to as Buchnera), which is housed inside bacteriocytes and is strictly vertically transmitted from the mother to offsprings , reproduction (Wolbachia), body color (Rickettsiella), host plant fitness (Regiella insecticola), heat shock enhancement, parasitoid as well as pathogenic resistance   .Bemisia tabaci, which can feed on over 600 plants species, including vegetables, fibers, and ornamental crops, is one of the most destructive insect pests. It is a natural vector of persistently transmitted begomoviruses (family Geminiviridae) as well as some semi-persistently and non-persistently transmitted plant viruses , the obligate endosymbiont for all whitefly species, is localized in the bacteriome, which comprises several bacteriocytes. It provides essential amino acids, carotenoids, and other metabolites that are scanty in a phloem sap diet to its insect host , such as leafhoppers, treehoppers, cicadas, planthoppers, and spittlebugs, are ancient and complicated. The obligate symbiont, Sulcia muelleri (hereafter referred to as Sulcia), and one or two co-obligate betaproteobacterial symbionts are associated with many hosts  to provide essential amino acids  and the Rickettsia non-infected (Rick–). They showed that Rickettsia infections enhance tomato yellow leaf curl virus (TYLCV) acquisition, retention, and transmission by insects  infection by enhancing the expression levels of antimicrobial peptides in gut epithelial cells  and revealed that PLRV binds the symbionin expressed by the obligate endosymbiont of M. persicae, Buchnera   mutants devoid of the readthrough domain (RTD) exhibited the same stability as wild-type viruses in aphid hemolymphs . This in in vivo . Further in vivo . ThroughYDV-RPV) . They prsmission . Taken tHamiltonella, a facultative endosymbiont of the whitefly, showed 80% homology with that from different aphid species and GroEL from E. coli. Feeding the anti-Buchnera GroEL antiserum to B. tabaci whitefly before viral acquisition inhibited TYLCV transmission by more than 80%  and Hamiltonella non-infected (H–)] from a same genetic background insect strain via antibiotic treatment and introgression. Through further studies, they found that whiteflies harboring Hamiltonella were suitable for TYLCV acquisition, retention, and spread (Hamiltonella on whiteflies has been associated with the transmission efficiency of non-circulative viruses (Arsenophonus (another facultative symbiotic bacterium of whitefly) was also found to interact with the Cotton leaf curl virus in vitro and in vivo  that showed interactions with GroEL, but not to grapevine virus A (GAV) or tobacco mosaic virus (TMV) that did not show any interactions , which is detrimental to aphid fecundity , belonging to the genus otein P8 , 1998. Td Nasuia . Besidesractions . RDV virvesicles . In the ic space . In thisRickettsia has been shown to promote TYLCV acquisition, retention, and transmission through insects  has been stably introduced into brown planthopper, Nilaparvata lugens and shown to inhibited infection and transmission of Rice ragged stunt virus (RRSV) and mitigated virus-induced symptoms in rice plants (In summary, horizontal and vertical transmission of persistent plant viruses by vector insects involve complex interactions among viruses, vector components, and symbionts. In recent years, symbiotic bacteria that regulate the spread of persistent plant viruses through its insect vector have been extensively studied. For instance,  insects , GroEL memolymph , Sulcia neration . Elsewheneration . Overalle plants .Although knowledge about the interplay among symbiotic microorganisms, insect vectors, and plant virus has rapidly expanded, there are several issues need further investigation. For instance, intestinal microbiota have been found to modulate immune responses, influence the physical status, or directly utilize microbiota-derived products to promote or inhibit arbovirus infection . Are theWW, H-WS, and QM: conceptualization. WW and QM: writing original draft preparation. WW, H-WS, J-ML, C-XZ, J-PC, and QM: writing, review, and editing. All authors have read and agreed to the published version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Carcass grading is a vital process in the slaughterhouse and is used for the quantification of the overall value of carcasses. Since carcass grading is often performed manually by a team of grading experts, it is subject to human limitations which result in inconsistency and limited operation speed. Considering this, an automatic beef carcass yield estimation system capable of predicting 23 key yield parameters was developed. However, just like any freshly introduced system, analysis of the economic impact of the grading system is vital before deployment in any slaughterhouse business. In this study, a thorough economic analysis to justify deploying the developed beef carcass grading system in a standard slaughterhouse in South Korea was performed through a cost-benefit analysis. The analysis found that the benefits derived from using the developed system outweigh the costs of purchasing and operating the system making the endeavor economically viable.To minimize production costs, reduce mistakes, and improve consistency, modern-day slaughterhouses have turned to automated technologies for operations such as cutting, deboning, etc. One of the most vital operations in the slaughterhouse is carcass grading, usually performed manually by grading staff, which creates a bottleneck in terms of production speed and consistency. To speed up the carcass grading process, we developed an online system that uses image analysis and statistical tools to estimate up to 23 key yield parameters. A thorough economic analysis is required to aid slaughterhouses in making informed decisions about the risks and benefits of investing in the system. We therefore conducted an economic analysis of the system using a cost-benefit analysis , internal rate of return (IRR), and benefit/cost ratio (BCR)) and sensitivity analysis. The benefits considered for analysis include labor cost reduction and gross margin improvement arising from optimizing breeding practices with the use of the data obtained from the system. The cost-benefit analysis of the system resulted in an NPV of approximately 310.9 million Korean Won (KRW), a BCR of 1.72, and an IRR of 22.28%, which means the benefits outweigh the costs in the long term. The livestock industry accounts for approximately 40% of agricultural production in Korea . LivestoFor a reliable and consistent supply of beef to satisfy the consumer market, there is a need to modernize slaughterhouse operations. For this reason, many slaughterhouses use an overhead rail system, which allows for linear and rotational movement of the carcasses, enabling bulk processing . HoweverBefore the beef is sent to market, beef grading is a key requirement of the production pipeline . Due to Initially, beef carcass grading was based on visual assessment, which was found to be subjective and inconsistent. This led to the development of automated video image analysis (VIA) technology in the 1980s. VIA technology involves taking images of the carcass and using software to predict quality-related features ,11, magnAlthough these technologies are helpful in the prediction of carcass yield characteristics, most are time-consuming, costly, and have low accuracy in the prediction of key yield parameters. Their low performance could be due to limitations in data/image acquisition systems, outdated and insufficient image and data analysis methods, and a limited sample size. These limitations formed the background for the development of an online image analysis-based system for the prediction of beef yield and quality.The developed system takes advantage of recent image analysis and statistical tools to estimate up to 23 yield parameters (including carcass lean meat percentage (LMP), major cut weight, hot carcass weight (HCW), etc.) with image data captured from two cameras giving medial and dorsal views of a beef carcass after slaughter.To justify the financial viability of this system in a slaughterhouse, it is important to determine its true value and the benefits and costs associated with its adoption. This emphasizes the importance of a thorough economic analysis to ascertain the importance of every variable subjected to the analysis. Therefore, good strategies are vital in the determination of the items to be considered as costs or benefits, the time periods in which they occur, and the analysis methods employed. Some of the methods for economic analysis of systems include a cost-benefit analysis (the most widespread), a cost-utility analysis, a risk-benefit analysis, and an economic impact analysis.The cost-benefit analysis (CBA) of an investment is a decision-making tool based on forecasts of variables over a predefined period. These variables from a CBA are affected by various factors whose effects need to be evaluated to determine the viability of the investment, achieved by carrying out a sensitivity analysis. A sensitivity analysis is the process of recalculating outcomes under alternative assumptions to determine the impact of an input variable; it is considered critical to model validation . A sensiIn order to determine the viability of a project, all aspects  must be expressed in terms of monetary value. In the cattle industry, CBA has been applied in the development of a model to investigate the economic performance of Scottish beef enterprises , the detThis study was therefore conducted to determine the economic viability of the introduction of this novel system into the beef grading pipeline in a standard slaughterhouse  in South Korea. This study examines the rigors of determining the expected present and future costs, the benefits of procuring the system, and how they feature over the lifespan of the system to provide an informed basis for investment decision making.The economic analysis process involves the evaluation of costs and benefits, which help us to rank projects based on economic viability for the better allocation of resources. It aims at analyzing the economic and welfare impact of a project . EconomiThis economic analysis was performed through a series of steps presented in this section. These include determining the project life, discount rate, enumeration of benefits, and costs of procuring the system for a typical slaughterhouse in South Korea. This is followed by the actual CBA and a sensitivity analysis.The beef carcass yield estimation system developed seen in a consistThe implementation of this system has multiple advantages including a reduction in the number of and/or support for carcass grading technicians, which saves costs and helps ensure the consistency of grading results. The system also provides yield data before actual carcass dissection and can be used to build an online auction database for order management. It results in reduced risk of the contamination that arises from carcass handling since the system is noncontact, and an improvement of yield through breeding optimization using data from the system. Despite the system’s ability to predict yield parameters with a commendable degree of accuracy, it cannot completely replace the grading staff, as some key grading parameters such as marbling still need to be evaluated from the analysis of the rib-eye in Korea (between the 12th and 13th ribs).To determine the lifetime of a project, it is important to determine how long the project is expected to last. The project’s lifetime is determined by how long until an asset/equipment generates more income than it costs to maintain and operate, i.e., how long the net economic value of an asset takes to be reduced to zero. According to the Korean Public Procurement Service (PPS), the persistent period of similar industrial equipment/machinery machines was set to nine years , and thuThe discount rate is defined as the interest rate used to calculate future cash flows to their present-day value. It is used in the computation of the time value of money, which is instrumental in a CBA, specifically for the calculation of NPV and IRR. According to the Korea Development Institute (KDI) recommendations for evaluating public projects, the discount rate value was set to 4.5% for this project .The cost is the monetary value of expenditures for supplies, services, labor, products, equipment, etc. that are not available for use anymore. This subsection breaks down the costs that arise from using the system. In a cost-benefit analysis, costs are divided into fixed and variable costs. In the context of this study, fixed costs refer to predetermined expenses that remain the same throughout the project life, regardless of production output, while variable costs refer to expenses that vary directly or proportionally depending on the production output of the project.The fixed costs considered for this project include the initial price of the system, installation costs, and the cost of the space where the system is to be installed. These costs will be incurred once at the start of the project rather than throughout the project life.The initial price of procuring the system was internally determined by the system development team. This was determined based on the cost of individual system components, research and development, and system software development. The price of procuring the system was set to 79,500,000 Korean won (KRW), which is about 16% of the average cost of the most renowned image-based carcass evaluation technologies , BCC-3 , and DEXA . This makes the system significantly cheaper than the alternatives while having relatively good prediction performance, albeit for fewer yield parameters.The cost for installing the system includes all the costs associated with system component setup and calibration, and overhead rail modification to suit the system design. The process of system component setup and calibration was assumed to be accomplished by a team of two technicians for a duration of one week. Based on the average daily wage of technical workers (with expertise in system software development) in South Korea, the cost of system component setup and calibration was determined to be 2,601,210 KRW. The cost for overhead rail modification based on the system design b was det2 (3.5 m × 5 m) in the slaughterhouse. To compute the cost of this space, the locations of 20 slaughterhouses in South Korea were considered in order to determine the unit cost of land (price per m2) in these locations [2 was then multiplied by the system space requirement. The total cost of the land occupied by the system was determined to be 3,441,498 KRW.From ocations . The aveThe variable costs considered for this project included system operational costs and system repair and maintenance costs. Most variable costs were incurred annually, except for some repair and maintenance costs, which followed the individual component warranty period.The operational costs incurred using this system include electrical costs, internet costs, and administrative costs. The electrical costs were computed based on the total system power consumption (approximately 860 W), system operation time, and the annual cost of electricity, which was computed using Equation (1) below. The system operation time was set to 9 a.m.–12 p.m. and 1–6 p.m., which is a total of 8 h per day. The type of electricity consumed by the system was based on the type of electricity consumed by an average scale of the slaughterhouse (consumption: 300 kW or more) in South Korea, which is Industrial B, high-voltage A, option II, as defined by Korea Electric Power Corporation (KEPCO) [The details of the calculations are that summer lasts from June to August (with two peak and six mid-peak hours), winter from November to February (with five peak and three mid-peak hours), and spring and fall from March to May and September to October, respectively (with two peak and six mid-peak hours).Since the system stores the images and grades results on NAS, which is accessible by the technical team via the internet, an internet connection with sufficient speed and reliability is required. For this reason, the average internet usage fee from three Korean mobile internet service providers  was sourced. The average internet rate for each mobile service company differed depending on the speed of the internet. For this study, a 100 Mbps internet connection was selected since it is sufficient, and the annual cost was found to be 270,000 KRW .Administrative costs are the costs incurred by a company that are not directly related to manufacturing but are relevant to the overall profitability of the company. The administrative costs for this project mostly relate to system management  and information technology services. The administrative costs for this project were set to 15% of the fixed costs of the project, amounting to approximately 12,624,009 KRW.The repair and maintenance costs include the repair and replacement of core system components , software support, and replacement of RFID tags.The core system components such as the PC and the NAS are to be repaired in the fourth and eighth years of the project, a decision based on the manufacturer’s product warranty. The cost of repairing the PC and NAS was set to 25% of their initial cost, resulting in 489,930 and 433,000 KRW, respectively, for each repair period. The cameras had a warranty of four years; therefore, camera replacements were planned in the fourth and eighth years of the project. The cost of replacing the cameras was 6,299,593 KRW for each replacement period. Because of possible exterior and/or interior damage to the RFID tags, 10% of the RFID tags (80 tags) were replaced every year. RFID tag replacement cost 641,200 KRW per year based on a price of 8015 KRW for every tag . A summaDepreciation refers to the process of allocating an amount less the residual value from the acquisition cost of an asset in a systematic and reasonable manner during the period in which the asset is in operation. It is used to reallocate the cost of a tangible asset over its useful life span. The depreciation costs were calculated using the sum of the years’ digits (SYD) depreciation method . The SYDA benefit is an advantage or profit gained from something. This subsection details the benefits that arise from using the system. For this study, benefits are divided into direct, indirect, and intangible benefits. Indirect benefits are returns from using a system or asset that cannot be directly observed but are nonetheless realized, while direct benefits can easily be observed and quantified.Through the utilization of the system, there is a direct reduction in the burden and fatigue of grading carcass and consequently a reduction in labor. The analysis assumes that, for an average Korean slaughterhouse with two or more grading experts, the introduction of the system will reduce the size of the grading team since yield estimation, a key step in carcass grading, will be performed by the system. It is assumed that the system will be able to directly benefit the slaughterhouse by saving the salary of at least one person from the grading team. This will result in a benefit of 42,509,220 KRW per annum based on the average wages earned by a middle-class quality control worker in South Korea .In South Korea, there are currently over 93,000 beef cattle farmers and over 3.5 million head of cattle. According to statistics, about 43% of the cattle are owned by largescale farmers who own over 100 heads of cattle each . It alsoThe system can be used to estimate carcass yield parameters and subsequently share the yield information with the livestock farmers to be used as an evaluation basis for the optimization of the breeding process. This can result in the production of animals with good meat yield, for a shorter growth time, at an optimized breeding cost for the farmer.Our analysis follows the breeding and slaughter system in South Korea, where farmers grow cattle to full maturity before choosing from three pathways for slaughter. Pathway one (practiced by 4.2% of farmers) involves the farmer paying the slaughterhouse to slaughter, inspect, and grade their cattle for a price of approximately 170,000 KRW, after which they sell the carcass to the highest bidder through auctioning. For pathway two (practiced by 45.9% of farmers), the farmer sells the cattle to a livestock broker who goes through a similar process before selling the meat or carcasses in an auction. Lastly, in pathway three (practiced by 49.9% of farmers), the farmers sell the cattle to a large corporation that also goes through the same process before finding the most suitable market to sell the meat ,41. In tTo enumerate the benefits resulting from breeding practice optimization due to the shared information between slaughterhouses and cattle farmers, we made the following assumption. According to the Korean Ministry of Agriculture livestock improvement goals , the raty 688 kg . BecauseThe utilization of our system to estimate carcass yield and the subsequent sharing of the yield data with farmers can result in further breeding practice optimization and a reduction in the unit production cost. The resultant unit cost saving was assumed to be a maximum of 0.5% throughout the project life, which would not be significant in the initial years of the project but would increase to a maximum of 0.5% in the final years following a custom sigmoid function . The difer annum , computeFor this study, intangible benefits consist of benefits that directly or indirectly result from the utilization of the system for carcass yield estimation but are difficult to quantify in monetary terms. These include improvements in worker safety and food safety, the creation of an online database, etc.The introduction of the system to the slaughterhouse would simplify the availability and accessibility of the carcass image data and resultant yield information. The readily available carcass data can be used to create an online database that makes information available to stakeholders  so they can make informed decisions or build systems upon the data. In Korea, this information can be used by the government to monitor national carcass production in real-time. It can also be used to update the online auction system (currently under implementation in Korea ), thus eSince the system is noncontact in nature, yield parameters that used to be determined in a hands-on manner can now be assessed automatically. Therefore, the system can reduce the food safety issues that may arise from slaughterhouse handling. Additionally, there is a reduction in workers’ exposure to dangerous tools and equipment , thus improving worker safety.The purpose of a CBA is to determine if the decision to invest in a system is sound and to know if the resultant benefits outweigh the costs, and by how much. All the flows of benefits and costs over time are expressed in terms of their NPV, regardless of whether they are incurred at different times. The CBA techniques considered for the evaluation of the economic feasibility of the system include NPV, BCR, and IRR.Net present value (NPV) analysis is commonly used for evaluating the feasibility of a project. Because the costs and benefits arising from a project continue into the future, future benefits and costs should be considered by converting them to present values. In simple terms, NPV refers to the present value of the expected future benefits and costs resulting from the investment . The forBCR refers to the ratio of the present value of the benefits  is a metric used in financial analysis to estimate the profitability of potential investments. It is defined as the discount rate that makes the NPV of the project zero. Generally, the higher the IRR value, the more desirable an investment is to undertake. If the IRR is greater than the discount rate, the investment is economically valuable. The computation of IRR was performed using Equation (8):A sensitivity analysis investigates how changes in the assumptions of an economic analysis generally affect the predictions. Ideally, the determined benefits and costs in the cost-benefit analysis include many assumptions and uncertainties. Therefore, a sensitivity analysis is performed to address these uncertainties. The sensitivity analysis identifies changes in the NPV, IRR, and BCR values by changing the critical variables that can affect the economic value within a certain range. For this study, five parameters were considered  for the economic analysis to be sufficiently sensitive. These parameters were varied while monitoring the changes in NPV, IRR, and BCR values. Further details of the variation of the parameters for the sensitivity analysis are presented in The results of the economic analysis, including the resultant cash flow and CBA, are summarized in From the results in On further analysis of the present value of the project cash flows over the project’s useful life, it was found that the break-even point (the point at which the project turns profitable) was achieved during the sixth year of the project . When the initial investment was increased by 20%, the values of NPV, BCR, and IRR changed to 292.2 million KRW, 1.65, and 19.27%, respectively.The benefits of the introduction of this system in a Korean slaughterhouse are a reduction in labor costs and subsequent optimization of breeding practices. An increase in both benefits resulted in increases in the NPV, BCR, and IRR values because they directly reduced the costs and increased the benefits of the implementation of the system. When no employees were replaced and costs were saved due to breeding practice optimization being maintained at 0.5%, the project achieved NPV, BCR, and IRR values of 2.0 million KRW, 1.01, and 0.14%, respectively. Conversely, when the benefit of costs saved due to breeding practice optimization was set to 0%, the NPV, BCR, and IRR values were −122.0 million KRW, 0.72, and −38.02%.Lastly, the effect of increasing the average annual carcass throughput of slaughterhouses resulted in an increase in the values of NPV, BCR, and IRR and vice versa.From the results of the project cash flows and CBA as summarized in While the benefits from the project generally increased throughout the project’s life, the rate at which the benefits grew was low in the early years, improved in the middle years of the project, and slowed down when nearing the end of the project. This was mainly because of the choice to reflect the indirect benefits (due to the breeding practice optimization resulting from using the data from our system) to follow a sigmoid function (described in Equation (3)).From the CBA, the project resulted in a positive value of NPV (approx. 311 million KRW), a BCR of 1.72, and an IRR of 22.28%. Since the achieved value of NPV was positive, BCR > 1, and IRR was greater than the nominal discount rate (4.5%), this means that the benefits far outweigh the costs arising from using the system thus investing in the system would be a good investment with a positive economic return.Since the project was observed to hit break-even in the 6th year of the project, this further emphasizes the profitability of the project and can be used as a basis to persuade investors.The sensitivity analysis results showed that the decrease in discount rate resulted in increases in the NPV, BCR, and IRR values and vice versa. Lowering the discount rate resulted in an increase in the net present value of the project. This is because most of the benefits of the project increased towards the end of the project life, thus the present value of such benefits was quite high compared to the costs.The improvement of the NPV, BCR, and IRR values on reduction of the initial costs of investment was because of the reduced costs due to the lower initial investment and reduction on depreciation costs.The effect of varying the project benefits directly affects the NPV, BCR, and IRR values. On reduction of the project benefits, the NPV, IRR, and BCR values reduce because this directly reduces the net present benefits of the project, the same happens vice versa. Therefore, an attempt to eliminate any of the project benefits resulted in the project not being viable economically.The reason for the increase in the values of NPV, BCR, and IRR with the increase of the average annual carcass throughput of slaughterhouses is because the increase in carcass throughput directly contributes to the benefit of costs saved due to breeding practice optimization. To investigate further the combined effect of carcass annual slaughter volume and costs saved due to breeding practice optimization on the values of NPV, BCR, and IRR was investigated. It was observed that increasing the annual slaughter volume (in steps of 350 carcasses) was not as impactful as increasing the costs saved due to breeding practice optimization. This implies that, for the system to achieve maximum economic impact, the annual slaughter volume should be increased above the average Korean slaughterhouse annual volume. Most importantly, emphasis should be put on sharing carcass yield data with livestock farmers to optimize breeding processes, reduce unit production costs, and consequently increase production cost savings for the farmers.A comparison of the impact of the five selected variables on the overall CBA is given in This study was conducted to determine the economic value and utility of an online image analysis-based beef carcass yield estimation system using a cost-benefit analysis. During the persistent period of this system, the net present value was found to be 310.9 million KRW, the BCR was 1.72, and the IRR was 22.28%. In each of the economic analyses undertaken, the benefits outweighed the costs in the long term. In particular, the investment was predicted to pass the break-even point within four years due to the benefits. As the sensitivity analysis suggests, to maximize the economic effects of the system, the number of cattle slaughtered should be increased. Most importantly, the grading data of carcass yields should be shared with the breeding farmers and be used for breeding optimization, which reduces the production costs.Since the CBA in this study was performed based on assumptions, there are some limitations that require attention and deeper analyses. Assumptions made in the determination of persistent time, discount rate, system capabilities, breeding optimization, etc., could easily bias the analysis and deem it unrealistic. For this reason, comprehensive research on the variables needs to be undertaken and a thorough sensitivity analysis is required. While the research and sensitivity analysis performed for this study was acceptable, there is still room for improvement in further studies.The yield results from the system can be used to build a useful database that is accessible to stakeholders and plays a vital role in decision making. This database can be used, for example, by governments to monitor carcass yield across the nation, to feed into the online auction system, predict beef carcass yield estimates prior to any further dissection, etc.This study proves that the automation of the slaughter line is economically viable and can be employed for modernizing meat production in Korea. Since this is developed domestically, the initial costs of the system and the repair and maintenance costs are relatively low in relation to other renowned imaging-based carcass grading systems. As proved by an earlier economic analysis of an imported pork carcass grading system , it can