Sodium hypochlorite (at 10%) and ethanol (at 80%) reduced the titre of Nipah virus to undetectable levels. Nipah virus that was in tissue culture medium was also inactivated after 24 h in the presence of 10% formalin.Nipah virus is a relatively newly discovered emerging virus on the WHO list of priority pathogens which has the potential to cause outbreaks with high fatality rates. Whilst progress is being made in the development of animal models for evaluating vaccines and therapies, some of the more fundamental data on Nipah virus are lacking. We performed studies to generate novel information on the aerosol survival of Nipah virus and to look at the efficacy of two common disinfectants. We also performed studies to evaluate the inactivation of Nipah virus by using neutral buffered formalin. Nipah virus was relatively stable in a small particle (1–5 µm) aerosol in the dark, with it having a decay rate of 1.46%min Nipah virus (NiV) is a zoonotic RNA virus of the Paramyxoviridae family and the Henipavirus genus . A relatNiV is on the WHO High Priority pathogens list which means it is amongst those “diseases that pose the greatest public health risk due to their epidemic potential and/or where there is no or insufficient countermeasure”, and it is therefore in need of urgent Research and Development action . As the In this paper, we present novel data on the survival of NiV in small particle aerosols and demonstrate the efficacy of bleach and alcohol-based disinfection through the testing of reduction in NiV titre after short contact times with a range of concentrations of sodium hypochlorite and ethanol. We also present our data on the inactivation of NiV by neutral buffered formalin which has enabled us to maximise data from in vivo studies by allowing the safe handling of formalin-fixed tissues from animals that are infected with NiV.6 TCID50/mL. The TCM was Dulbecco’s minimum essential media  which was supplemented with 2% Fetal Calf Serum, 1% L-glutamine and 1% Penicillin/streptomycin . Nipah virus-Malaysia (hereafter referred to as NiV) was kindly provided by Viral Special Pathogens Branch, (CDC) where it was derived from the “199901924” isolate and had undergone 2 passages in Vero-E6 cells and one passage in RK13 cells. At Dstl, the virus was passaged a further two times in Vero C1008 cells (ECCAC Cat. No. 85020206), and it was maintained in the tissue culture media (TCM) to produce a working stock of 2 × 1050) assay to enumerate a viable virus as described previously [2 for 5–7 days and observed under a microscope for the presence of cytopathic effects (CPE). Each column of 8 wells of cells was scored for the presence or absence of CPE and the 50% end-point was calculated by the methods of Reed and Muench [NiV was titrated by a 50% tissue culture infectious dose  Ltd., Milton Keynes, UK). For the passaging of samples, flasks of increasing size were used for sequential passage. All of the flasks  had a confluent monolayer of Vero C1008 cells which was maintained in TCM that was supplemented with 2% Fetal Calf Serum, 1% L-glutamine and 1% Penicillin/streptomycin. The final volumes in the flasks were 5 mL (T12.5 flasks), 10 mL (T25 flasks), 20 mL (T75 flasks) or 30 mL (T150 flasks). The flasks were incubated at 37 °C and 5% CO50 assay in triplicate, making a total of 9 values for each sample point. All drum experiments were performed in the dark, at an ambient temperature (19–21 °C) and a relative humidity (42–57%). In separate validation experiments, Bacillus atrophaeus (BA) spores were aerosolised under the same conditions as the virus was, as is described above. The BA spores at 108 cfu/mL were diluted 1:10 in Phosphate Buffered Saline  prior to aerosolisation. The impinger samples were collected in a PBS, and viable counts were performed on L-agar plates. For the aerosol data analysis, each time point value was corrected for the dilution effect of removing the air. The data was log10 transformed, and a best-fit line was determined using GraphPad Prism v 8 . Studies to determine virus survival in small particle aerosols were performed using the rotating 40 L Goldberg drum as described previously ,10,11, w50 assay was performed on a 1:10 dilution (0.1 mL) of the pellet, and the remaining 0.5 mL was inoculated into flasks and processed as above. The entire inactivation process was repeated on three further occasions. Ultra-centrifugation was used to remove formalin from the samples. For each round, 1 mL 20% formalin (Sigma) was added to 1 mL NiV virus stock, in triplicate . Two positive control samples were made by adding 1 mL TCM to 1 mL NiV virus stock, and the final sample was a negative control of 1 mL 20% formalin + 1 mL TCM. All samples were incubated at room temperature for 24 h. After 24 h, 30 mL TCM was added to each aliquot in ultracentrifuge tubes, then, 3 mL 30% sucrose was dispensed beneath the media to form a sucrose cushion. All of the samples were ultra-centrifuged at 25,000 rpm for 3.0–3.5 h at 4 °C (Beckman Optima Ultracentrifuge with SW28 rotor). After centrifugation, the supernatant was discarded, and a second wash was performed in the same manner as described above. The final pellets of the formalin-treated samples were re-suspended in 0.4 mL TCM. The entire sample (100%) was added to a T75 flask of confluent Vero C1008 cells in 20 mL media. The flasks were incubated at 37 °C for 6–7 days and then the entire contents of each flask were passaged onto fresh Vero C1008 cells in a T150 flask in 20 mL fresh media . The T150 flasks were incubated at 37 °C for 6–7 days and then the cell monolayers were observed using a light microscope, and the presence or absence of cytopathic effects were recorded by two individuals each time. For the production of positive control samples, the pellet, after two rounds of ultra-centrifugation, was resuspended in 0.5 mL TCM. A TCID2 in aqueous solution)  and ethanol  were prepared at a range of percentage amounts  in tap water, based on the undiluted, supplied reagent being equal to 100%. Disinfectants were prepared at 2× concentration so that when they were mixed at a 1:1 ratio with the virus, they were at concentrations of 40%, 30% or 20% ethanol (v/v) or 10%, 1%, 0.5% and 0.1% (v/v) sodium hypochlorite, respectively. Sodium hypochlorite (supplied as 14% Cl6 TCID50/mL) in a polystyrene 24-well culture plate . For the production of positive controls and to determine the maximum amount of virus recovery, 50 µL TCM was added to 50 µL NiV stock. The samples were tested in at least duplicate on two or three separate occasions. After 1 min or 5 min, 1 mL TCM was added to the virus + disinfectant, and then the samples were centrifuged using a microcentrifuge at 10,000 rpm for 5 min. After centrifugation, the liquid was discarded and 1 mL fresh TCM was added to the tube. This was repeated twice. After the final centrifugation, 1 mL TCM was added to the tube and a TCID50 assay was performed on each sample. The centrifugation steps were required to dilute the chemical components that would otherwise cause toxicity in the cell-culture based enumeration assay, and this method is similar to the methods that have been described previously [To test the virucidal activity of the disinfectants, 50 µL of the disinfectant (either ethanol or sodium hypochlorite) was added to 50 µL NiV stock . Based on this decay rate, the half-life of NiV in the drum is 47 min . Under these experimental conditions it would take 2.5 h (153 min) for the starting titre of virus to drop by 90% (a factor of 10).Small particle aerosols containing NiV were generated and held in the dark in a Goldberg drum over 90 min at ambient temperatures and a relative humidity. Impinger samples were collected at the following time intervals: 0, 5, 15, 30, 60 and 90 min, and were enumerated by a TCIDnsformed . Non-lin50 assay in triplicate. Three separate aerosol experiments were performed. Counts were adjusted for the dilution effect of sampling. The mean adjusted count of virus ± SEM and a best fit line from all of the runs are plotted. The limit of detection (LoD) and the limit of quantification (LoQ) of the TCID50 assays are shown as dotted lines.NiV was aerosolised in TCM and held in a Goldberg Drum at a medium relative humidity (50–60%). Impinger samples were taken at set time points and enumerated by TCID−1, 95% confidence intervals for k = −0.02672 to 0.007809). Taking into account the physical decay in the drum, the biological decay rate of NiV would be 1.16 %min−1.. This would then translate to a half-life of 60 min and it would take 198 min (3.3 h) for the titre to drop by 90%. The validation studies using BA spores in the drum system that was described above indicated that there was minimal physical decay in the system  reliably caused infection, whereas the flasks that were infected with a −7 dilution of NiV (approx. 0.2 TCID50) did not cause infection. This suggests that the LoD in the flasks is very low, at <2 TCID50 (results not shown). The limit of quantification of the TCID50 assay is 10 TCID50 (a result of 50% of the wells in the column containing a neat sample being infected). The limit of detection of the TCID50 assay is extrapolated to 3.73 TCID50 (based on a single well that was positive for infection in the column containing neat samples), therefore, the flasks are likely to be more sensitive in detecting a viable virus than the assay.NiV was diluted 1:10 down to a −8 dilution, and all of the dilutions were used to infect T12.5 flasks. The flasks were passaged three times. The flasks that were infected with a −6 dilution of NiV stock , and then they were tested for viral inactivation by infecting the cells with 100% of the sample which was followed by a subsequent passage. The starting titre of NiV was 2 × 10NiV was inactivated by 10% formalin  after 24 h incubation at room temperature. This method was reproducible; a total of 12 replicates were tested (three each time on four occasions). An example of the final cell monolayers is shown in v/v) and sodium hypochlorite at 10%, 1%, 0.5% and 0.1% (v/v) against NiV were tested for a contact time of 1 or 5 min. The samples were not mixed intentionally, but they did often coalesce to one large droplet of liquid. After the 1- or 5-min contact time, the samples were washed three times to remove the toxic components. The effects of ethanol at 80%, 40%, 30% and 20%  were able to reduce the titre of NiV to un-detectable levels by TCID50 assay (v/v) was also very effective against NiV; in two-thirds of the replicates that were tested, no viable virus was recovered after 1 min, and in the remaining third of the samples, the titre of the virus recovered was at the limit of detection of the assay. The two samples that resulted in a viral titre appeared in those flasks from the second passage; the remaining samples were negative throughout the passages. No NiV was detected by TCID50 assay or in the flasks when the contact time was increased to 5 min with 1% sodium hypochlorite  were still recovered from some replicates  which are known to have an impact on viral aerosol survival ,20, may 4 TCID50 NiV. The use of ethanol, at all other concentrations that were tested, resulted in some level of viable virus being detected in at least one of the replicates, even when the contact time was increased to five minutes. For sodium hypochlorite, 5 min contact time with 1% and 0.5% was effective. Under those conditions, where no viable virus was recovered, a 4 log10 reduction in the viral titre was observed, which is considered the standard measure for disinfectant testing [As of August 2022, there are no published data on NiV disinfection with sodium hypochlorite, and there is a single study that is focused on ethanol. . A 2019 review article reported that “NiV can be readily inactivated by soaps, detergents and commercially available disinfectants such as sodium hypochlorite” , but the testing , althougThe disinfection efficacy data that are produced in this study against NiV are comparable to the data that are published elsewhere for Ebola virus . In thosIn addition to sodium hypochlorite and ethanol, good disinfection efficacy has been achieved with a single concentration and a short contact time of several quaternary ammonium-based disinfectants against NiV (unpublished observations). Disinfection data provides useful fundamental knowledge for NiV disinfection that can be applied to laboratories that are performing research or in hospitals and clinical settings for people who are working with patients that are infected with NiV. As ever, with this type of work there are many more disinfectants, concentrations and contact times that could be investigated, and the work could also be extended to look at its efficacy in various matrices such as blood, where different parameters may be needed to achieve the same level of knockdown. The inactivation of NiV may be required for the exploitation of virus or virus-containing samples during research. Heat treatment at 56 °C or 60 °C has been shown to be effective at reducing the titre of NiV, as has UV irradiation for 30 min . The redIn conclusion, we report novel data on the survival of NiV in small particle aerosols, and on disinfection of NiV, which will help to inform risk assessments and control measures for those that are working with the virus in research or in outbreak situations. We have also added to data on the inactivation of NiV by neutral buffered formalin, which can be used to develop methods for exploiting samples from animals that are infected with NiV for histopathological analysis. This, in turn, will help in understanding disease pathogenesis and in the assessment of the efficacy of medical countermeasures in in vivo models.


A diagnosis of cancer is associated with significant physical, psychological and financial burden. Including costs of cancer is an important component of shared decision making. Doctors bear a responsibility towards educating patients about the financial aspects of care. Multiple organisations have advocated for price transparency and implementing Informed Financial Consent in the clinic. However, few studies have evaluated the perspectives of oncologists on the current state of this discussion.The aim of this study is to determine the views and perspectives of medical oncologists regarding communication of costs and financial burden in patients with cancer.We conducted a prospective cross-sectional online survey via REDCap. The survey was distributed to medical oncologists and advanced trainees currently registered with Medical Oncology Group of Australia (MOGA). Data was collected using the online survey comprising socio-demographic characteristics, discussion of costs and financial burden, and facilitators and barriers to these discussions.547 members of MOGA were invited to participate in the study, and 106 of 547 MOGA members (19%) completed the survey. Most oncologists (66%) felt that it was their responsibility to discuss costs of care, however a majority of oncologists (59.3%) reported discussing costs with less than half of their patients. Only 25% of oncologists discussed financial concerns with more than half of their patients, and most oncologists were unfamiliar with cancer-related financial burden. Most Oncologists with greater clinical experience and those working in private practice were more likely to discuss costs with a majority of their patients.Certain characteristics of medical oncologists and their practices were associated with reported prevalence of discussing costs of care and financial burden with their patients. In the context of rising costs of cancer care, interventions targeting modifiable factors such as raising oncologist awareness of costs of care and financial burden, screening for financial toxicity and availability of costs information in an easily accessible manner, may help increase the frequency of patient-doctor discussions about costs of care, contributing to informed decision-making and higher-quality cancer care. A diagnosis of cancer is associated with significant physical, psychological and financial burden. Costs of cancer care account for a substantial proportion of healthcare expenditure, and healthcare costs for cancer are higher than for other conditions . Newer aDoctors bear a responsibility for educating patients about the financial aspects of cancer treatment. Multiple organisations , includiFew studies have evaluated the perspectives and attitudes of doctors with regards to discussions of costs of care or financial burden. We aimed to describe the frequency and nature of both cost of care and financial burden discussions in a clinical setting, and to determine the characteristics of medical oncologists and their practices that are associated with discussions about costs. We also sought to identify strategies and potentially modifiable factors to facilitate future discussions about costs of care and financial burden.We distributed an online survey to medical oncologists and medical oncology trainees practising in Australia that were listed on a professional association database . MOGA is the national, professional organisation for medical oncologists in Australia. Participants were asked to recall discussions undertaken in the clinic over the last 12 months regarding the costs of cancer treatment, and the evaluation and management of financial burden. Clinical discussions included any conversations undertaken between clinicians and patients in outpatient clinics during a consultation. Socio-demographic characteristics of participants included: age, gender, qualifications, practice location, tumour streams, geographical region of practice, average number of patients seen per week, and practice type (public or private). The prevalence of discussions about costs of care was categorised as: with none of the patients, less than half of the patients, more than half of the patients, and all of the patients. These categories were dichotomised for analysis into, less than half of the patients, versus more than half or more of the patients. Discussions of financial burden were categorised in the same way.The oncologist survey was developed by three medical oncologists  and a health economist (RLM). Prior to wider distribution, the survey was piloted with two oncologists and two researchers to test clarity, content validity and face validity. The survey was delivered, distributed, and collected online using REDCap software.Descriptive statistics were reported to describe oncologist characteristics and the prevalence of discussions of costs of care, and financial burden. We quantified oncologist preferences for discussions of cost of care and financial burden using frequency distributions. To assess whether socio-demographic characteristics were associated with the frequency of discussions about costs of care (<50% vs >50%) and financial burden (<50% vs >50%), we built two multivariable logistic regression models. We reported unadjusted odds ratios (ORs), adjusted ORs, and 95% confidence intervals (CIs). First, we examined the univariate associations for each characteristic separately by examining the unadjusted odds ratios. We then built four multivariable regression models for each dependent variable in a stepwise approach. We included all characteristics of the oncologists, and then included only the characteristics that were significantly associated with the discussion of costs of care in the univariate analyses, and those that were relevant to the analysis including location of the oncology practice, average number of patients seen per week, and the type of practice. The final multivariable model included characteristics that were relevant to the discussion of out-of-pocket costs. Analyses were performed with SAS 9.4 and Graphpad PRISM 8 software.This study was reported according to the STROBE checklist for observational studies .547 members of MOGA were invited to participate in the study, and 106 of 547 MOGA members (19%) completed the survey. The majority of responders were qualified medical oncologists (83%), and approximately half were female (53%) . 78% praIn response to how often oncologists discussed costs of cancer care with patients in the past 12 months, 3.7% reported never discussing costs; 55.6% reported discussing costs with less than half their patients; 33.9% discussed costs with more than half their patients and 5.6% discussed costs with all their patients.Most oncologists (66%) felt that it was their responsibility to discuss costs of care, rather than that of other healthcare professionals in the clinic . Most , and genomic tests not subsidised by Medicare . OncologThe following clinical characteristics were associated with discussing costs of care with more than 50% of patients: oncologists with greater clinical experience, and those working in private practice. Oncologists with greater than 5 years of practice since attaining specialist qualifications were more likely to discuss costs of care compared with their junior counterparts OR 7.8  . OncologOncologists were asked to rate the influence of current and potential strategies that could facilitate discussions about costs of care in their clinics. Facilitators that were rated by oncologists as extremely helpful included: availability of cost information on a website (51% of oncologists), and handouts for patients with information about costs (28%) . StrategClinicians were also asked to rate the influence of potential barriers to discussions of costs of care during a clinical consultation. Oncologists reported lack of available information about costs (36%), and limited solutions for cost concerns (18%) as major barriers for discussing costs of care with patients . CliniciIn response to the question about how frequently oncologists discussed financial concerns with patients and/or their families, 4% reported never having discussed it, 71% that they had discussed it with less than half their patients, 22% with more than half their patients, and 3% with all their patients. Approximately half the oncologists (48%) reported that discussion of financial burden was generally initiated by patients, one-third (32%) reported the topic was usually initiated by doctors, and one-sixth (17%) that it was generally initiated by family members or carers, 3% that they were initiated by a member of the allied health team.Respondents reported limited knowledge of cancer-related financial burden, with most (93%) reporting little or no familiarity with a financial toxicity screening tool e.g. COST-FACIT , 13; litDespite this, approximately half of the respondents 56 of 106, (53%) had referred patients to financial assistance programs, financial resources, or counselling to assist with financial burden. A smaller proportion 30 of 106, (28%) had not made a referral themselves, but h relied on a different healthcare professional in their service to make appropriate referrals. One in seven respondents (14%) were not aware of any pertinent financial assistance programs or counselling, and 5 clinicians reported that they had no programs in their service to offer.Clinicians were asked to rate the helpfulness of current and potential strategies that could facilitate discussion of financial burden in the clinic. Most oncologists, 73 of 106, (69%) reported that the availability of a financial navigator or social worker in the clinic would be extremely helpful . Other sTools and resources reported to be most useful in assisting with communicating costs of care and financial burden were, a financial navigator in the clinic to discuss costs and resources with patients directly reported by 88 of 106 (83%), online healthcare cost information to assist clinicians in estimating expected costs of care reported by 72 of 106 (68%) and, online education modules on financial burden assessment and management reported by 42 of 106(40%). Resources that were less frequently found to be useful were an implementation pack for informed financial consent with financial burden screening tools and resources by 36 of 106 (33.9%), electronic medical record (EMR) flags or posters to prompt discussions of costs of care or financial burden with patients reported by 32 of 106 (30.2%), and interactive communication workshops on financial burden reported by 30 of 106(28.3%).This is the first survey of Australian medical oncologists regarding their practices and perspectives on discussions of costs of care and financial burden in patients with cancer. The study identified that oncologists thought it was their responsibility to discuss costs of care and financial burden with their patients. Oncologists who were qualified for more than five years, and those who worked in private practice, were more likely to report discussing the costs of cancer care with a majority of their patients than those who were early in their career (less than 5 years since graduation) or worked predominantly in public healthcare settings. We did not identify any oncologist characteristics associated with discussing financial burden. Approximately two-thirds of oncologists felt it was their responsibility to discuss of care, particularly out-of-pocket costs however, more than half of respondents reported discussing costs with less than half of their patients.In the past decade, there has been increased awareness of the rising costs of cancer care and financial burden for people affected by cancer. This has led numerous medical organisations in Australia and overseas to introduce standards for informed financial consent –5. In adThis study has limitations inherent to all surveys. The response rate was low but consistent with other studies that recruit through a professional association’s mailing list, and as with all surveys, there may have been differences between responders and non-responders. A majority of responders were from a single state , and although this is the most populous state in Australia, it is possible that there was greater awareness of cancer costs and financial burden due to the Cancer Council Financial Assistance programs .This study highlighted possible facilitators and barriers to discussing costs and financial burden from the oncologists’ perspective. We identified characteristics of oncologists and their practices that were associated with increased prevalence of cost of care discussions, including greater number of years since qualification and work in a private practice. A greater number of years in practice and higher patient volumes are known to be associated with aspects of cost-consciousness and awareness of patient out-of-pocket costs among clinicians . OncologThe findings of this study have significant implications for the future training and practice of medical oncologists regarding the implementation of cost transparency and assessment of financial burden. Despite the introduction of a standard for informed financial consent in medical oncology in Australia , cliniciIn addition, communication workshops and education modules outlining effective discussion of costs and financial burden can be implemented at different levels of doctor training from undergraduate degrees university to post-graduate qualifications. Clinicians feel a responsibility to discuss costs of care, however, they may face moral and ethical dilemmas regarding recommendation of unfunded or expensive treatments to patients with pre-existing financial burden . CliniciIn conclusion, we found characteristics of medical oncologists and their practices were associated with reported prevalence of discussing costs of care and financial burden with their patients. In the context of rising costs of cancer care, interventions targeting modifiable factors such as raising oncologist awareness of costs of care and financial burden, screening for financial toxicity and availability of costs information in an easily accessible manner, may help increase the frequency of patient-doctor discussions about costs of care, contributing to informed decision-making and higher-quality cancer care.S1 Data(CSV)Click here for additional data file.


The thoracic, lumbar, and hip angles and finger floor distance were measured to assess the change in ROM. Statistics indicated that the effect size in instantaneously change of flexion -and extension ROM by SKD was large . No significant effect was present in the sham condition. Flexion ROM decreased whereas the extension ROM increased, depending on SKD location- and displacement direction (p < 0.05). The ICC indicates a good intertester and intratester reliability . Lumbodorsal SKD affects the flexion- and extension spine, pelvis, and hip range of motion. The effects of SKD are direction- and location dependent as well as movement (flexion/extension) specific. Lumbodorsal SKD during flexion and extension may be useful to determine whether or not a patient would benefit from fascia tissue manipulations. Further research is required to obtain insight into the mechanisms via which the SKD affects ROM and muscle activation, in healthy, asymptomatic-LBP, and LBP subjects.Low back pain (LBP), associated with spine, pelvis, and hip mobility impairments can be caused by tight muscle contractions, to protect sensitized lumbar fasciae. Fascia tissue manipulations are used to treat lumbar fascia in LBP. The effect of fascia tissue manipulations through lumbodorsal skin displacement (SKD) on mobility is inconclusive likely depending on the location and displacement direction of the manipulation. This study aimed to assess whether lumbodorsal SKD affects the flexion -and extension range of motion (ROM), in healthy subjects. Furthermore, we aimed to test the effect of SKD at different locations and directions. Finally, to assess intertester and intratester reliability of SKD. Effects of SKD were tested in a motion capture, single-blinded, longitudinal, experimental study. Sixty-three subjects were randomly assigned to SKD- or sham group. SKD group was subjected to either mediolateral directed SKD during flexion or extension movement,  Low back pain is associated with spine, pelvis, and hip mobility impairments  hypothesIn physical therapy, “fascia tissue manipulation(s)” (FTM(s) are used in treating musculoskeletal pain, like low back pain. FTMs such as myofascial release techniques, myofascial trigger-point interventions, and elastic tape application methods have been applied and their effectiveness have been systematically reviewed . The effOptimization of FTMs requires a more detailed understanding of the underlying mechanisms of this type of treatment. It has been proposed that FTMs by displacement of the skin, the tension in the underlying fasciae will be modulated which alters the mechanical properties. In support of this rationale, mathematical geometric modeling has shown that forces exerted onto the skin can deform and displace the fasciae and as such change the mechanical properties of the underlying fasciae . As evid® Test. The test consists of 3 steps: 1) affirmation of the most painful movement from stance to either flexion or extension, as a reference test, 2) the same test with ongoing mediolateral directed SKD to the right at e.g., L3 or L5, and 3) the same reference test with ongoing SKD to the left. SKD leading to the largest mobility improvement and/or pain reduction can be utilized for FTMs at the tested location  . It has location .Several studies in which FTMs have been applied to healthy humans by elastic tape or myofascial release have shown that fasciae and muscles below the skin undergo deformations and are locally strained . Therefoversus a sham skin-displacement, in healthy subjects, and 2) if present, to test the effects of SKD at different locations and directions, as well as 3) to assess intertester and intratester reliability of applying the SKD.Although changes in joint mobility through SKD seem to be clinically effective, the basal effects of SKD on healthy subjects have not been tested objectively. Therefore, the aims of this study were: 1) to assess whether SKD at the lower back affects flexion- and extension range of motion of the spine, pelvis, and hip complex via advertisements that were placed on social media platforms (Facebook and Linkedin). Inclusion criteria were: Healthy subjects . We have chosen this age category because disability due to low back pain is highest or most severe at the age of 25–65-year  was performed. The following values were used for the expected effects of SKD: 1-β = 0.80, α = 0.50, effect size f2 = 0.15, 2*4, resulting in a minimum of 56 participants, n = 60) for possible dropouts, in line with the COSMIN  groups were created: an SKD- and a sham group. The stratified randomization method was used to secure homogeneity between the groups with regard to sex. The randomization was performed by a “blinded” observer, utilizing a computer-generated randomized table. Furthermore, each subject was randomly assigned to one of the four pre-selected orders of testing.th thoracic spinous process, and 4th thoracic spinous process. All markers were fixated to the skin by double-sided adhesive tape. All cluster-markers were additionally supported by an elastic band (Fabrifoam®) and Fixomull®strech tape . The three-dimensional positions of the markers were determined with an accuracy of up to 0.1 mm and resolution of 0.01 mm utilizing three Optotrak® cameras   to the pre-palpated anatomical landmarks, marked with a pencil by an experienced physiotherapist and four markers were attached to the custom-made station al Inc.)  at a samtal side .a priori experiment demonstrated no carry-over effects of the index tests when a 30-s pause was held between every single test. All above-described tests were repeated three times . The whole procedure was repeated twice with ongoing SKD applied by two separate testers (both experienced physiotherapists) to determine the SKD reliability through the agreement between two testers (1st vs. 2nd) and a third time by tester 1 to determine the within tester consistency (1st vs. 3rd), see Each subject started standing on a custom-made station. The custom-made station was designed in such a way that the knees could only maximally flex 10° through a leg-support. The degree of knee flexion was measured with a BASELINE®BUBBLE®INCLINOMETER. Depending on the randomization, at first, a maximal spinal flexion or extension movement (index test) was performed at own comfortable speed to retrieve the baseline reference value (repeated 3 times). In addition, either an ongoing lumbodorsal SKD or sham displacement was carried out, conform the four conditions (combination: location and direction) of the test procedure . The fouThe flexion- and extension range of motion (ROM) were assessed to estimate the mobility. The spine, pelvis, and hip complex were divided into 3 regions to analyze the thoracic, lumbar, and hip ROM changes  during flexion and extension. In addition, the distance between the wrist marker and 1st station marker was measured as the Finger Floor distance (FFD) . To calc−1). Before the data analyses started the raw data (C3D files) was displayed in Mokka©, a motion kinetic and kinematic analyzer, for evaluating the calibration process and marker acquisition (® data (NDF files) was used for data analysis utilizing a custom-made MATLAB script. Kinematic noise was filtered using a Butterworth Filter 4th order dual-pass with a cut-off frequency of 2.0 Hz. Ultimately, data was transformed into four-quadrant degrees (ROM changes were studied by determining the theta rotation (θ) inverse tangent  were determined by creating a tangent line from the cranial marker (C) to the medial marker (A), connecting medial marker (A) with the caudal marker (B) and creating a CAB vertex (∠CAB) . The CA versus sham on the thoracic, lumbar, and hip ROM (aim a), the interval data was transformed into absolute data (change or no change).To prepare the data for statistical analysis, the mean values were calculated for the three baseline tests and the three SKD tests. Furthermore, mean changes were calculated between TROM, LROM, HROM, and FFD concerning the mean baseline test per group and condition. To study the effect of SKD ® (version 27.0). An outlier labeling rule based on Interquartile Range with a 2.2 multiplier  . An a prversus sham, with 1 between-group variables (SKD group vs. sham group) and 4 within-group variables , at first using the absolute data (change or no change). Before the mixed-model ANOVA Mauchly’s test of sphericity was evaluated; an epsilon adjustment  was used due to a lack of sphericity. A Bonferroni post HOC test was used to evaluate the difference between SKD conditions per region. The magnitudes of the effect size for all conditions were calculated with the partial eta squared  95%.A mixed-model ANOVA was used, to study the effects of SKD on thoracic, lumbar, and hip ROM e effect . Interva3,k) was calculated and model 3,1 for intratester reliability  was calculated. An ICC below 0.5 was considered poor, an ICC between 0.5 and 0.75 as moderate, between 0.75 and 0.9 as good, and above 0.9 as excellent (The SKD reliability was determined by the intraclass correlation coefficient (ICC). Model 3,k for intertester reliability  were present Total seventy-five subjects were registered for the study. Twelve subjects were excluded for several reasons n = 1872; extension, n = 1,404). Thirty flexion and 46 extension data points were detected as extreme outliers and were excluded from analyses. The variables within each group had a normal distribution (p < 0.05). No significant carry-over effects were found regarding the order of testing (p < 0.05). The square root transformed data was back-transformed for interpretation.For one subject the HROM and LROM data points were excluded since the sacrum cluster was touched during the flexion measurements. In addition, for two subjects the flexion data were excluded due to exceeding the large but still limited range of motion in the experimental set-up. The processed data consisted of a total of 3276 records , not for LROM, TROM, and FFD (p > 0.05). No group and condition interaction effects were found for all regions (p < 0.05). For all regions, the mean flexion ROM was significantly greater in the SKD group (p < 0.05) p < 0.001). Detailed information regarding the SKD data is presented in Differences between SKD conditions per region are displayed in p > 0.05, ε > 0.75), hence, Huynh-Feldt correction was used for data interpretation. For all regions, the extension ROM was significantly greater in the SKD group (p < 0.05). SKD only affected LROM and TROM which was not different between SKD conditions. An interaction effect of group and condition was found for HROM . Within the SKD group, the affected HROM was dependent on the SKD condition applied . Post hoc analysis revealed that the HROM was differently affected between SKD conditions LL5 and LL3 (p < 0.05)  . No sign95% values (95% = 2.9), LROM (MDC95% = 1.5), and TROM (MDC95% = 0.9). In most SKD conditions a decrease in flexion ROM occurred for all regions, except for the thoracic which increased during SKD LL3, (p < 0.05)  .95%, extension HROM (MDC95% = 1.7), LROM (MDC95% = 1.4), and TROM (MDC95% = 1.6) increased in almost all SKD conditions. Contrary to the foregoing, a significant extension decrease was measured for the hip during SKD LL3 (p < 0.05) (Based on the MDC < 0.05) .p < 0.001) with an ICC3,k for all regions (0.81–0.93). The observed affected thoracic, lumbar, and hip ROM between the repeated SKD tests performed by a physiotherapist was consistent as shown by the ICC3,1 ranging from 0.70 to 0.84  and a decrease in extension ROM  due to SKD which was much greater than in the sham group. To the best of our knowledge, this is the first study showing that SKD affects the thoracic, lumbar, and hip ROM based on the SKD condition used. These findings support the hypothesis that skin and underlying fasciae are modulated by SKD. Consequently, this may have an impact on the structures that determine the thoracic, lumbar, and hip ROM. Whether this effect is due to altered tensions in underlying fasciae remains to be determined.This study shows that lumbodorsal mediolateral directed SKD affects thoracic, lumbar, and hip ROM. The effects of SKD are substantially direction -and location different as well as movement (flexion/extension) specific. In general, the mean flexion ROM decreased while mean extension ROM increased. However, based on the MDCn = 63), 5) the order of SKD conditions per group was randomized, and 6) the increase or decrease of ROM per region was based on surpassing the MDC95%.A strength of the study is the robust internal validity of the study because of several reasons: 1) the FDT consisted of a standardized procedure that was performed in a custom-made station in reducing the change of random errors , 2) the versus the sham group. Another limitation could have been that the subjects were unfamiliar with performing the index tests in a fixed position on an elevated station (20.5 cm), which could have increased fear. Fear is associated with increasing myoelectric activity of the lumbar erector muscles in healthy subjects  but also in the much lower normal stress (posterior-anterior pressure) used in the sham group. Previous studies have shown that the amount of pressure experienced by patients may influence the treatment effects  and alsosubjects  influencsubjects . This misubjects . The thosubjects . We haveases ROM . No carr® Test by evaluating the SKD effects on the spine, pelvis, and hip ROM  it is likely that amplitude, displacement, and force applied to the skin were fairly similar for the two experienced physiotherapists. It remains to be determined whether a difference in intensity between less and more experienced physiotherapists exists. The SKD is increasingly used by physiotherapists during physical examination to determine whether or not a patient would benefit from FTMs. However, this needs to be assessed. It is conceivable that for clinical purposes fascial diagnostic testing needs to be adapted and optimized. Moreover, whether the same effects of SKD occur in people with low back pain needs to be investigated.The initial SKD intensity aim was grade 4 . The quevia the lumbodorsal superficial fascia to the thoracolumbar fascia, myofascia, muscles, and thoracic, lumbar, and hip arthrofascia since they are linked via connective tissues  force during SKD is expected to be transmitted  tissues . These f tissues . This co tissues  which is tissues .In support of this rationale, most SKD conditions did not increase the flexion ROM and did not decrease the extension ROM. Subsequently, the affected flexion- and extension ROM differs between the SKD conditions. We hypothesize that changes occurs in: 1) fasciae position, 2) stiffness, and/or 3) agonist- and antagonistic muscle activity. Understanding the underlying mechanisms requires a different experimental design with other measurement instruments like ultrasonography and electromyography.This study shows that lumbodorsal SKD affects the spine, pelvis, and hip range of motion in healthy subjects. Flexion ROM decreased by SKD whereas extension ROM increased. The range of motion change depended on the SKD location and direction. The reliability of the SKD was remarkably good. These results suggest that the SKD may be a promising interventional test to obtain an indication, whether or not, a patient would benefit from FTMs. Further research is warranted to obtain insight into the mechanisms by which SKD affects the spine, pelvis, and hip range of motion, muscle activation, force transmission, in healthy, asymptomatic, and low back pain subjects.


The out-of-pocket costs to care for individuals living with dementia are high, and is exacerbated by the expense of managing behavioral and psychological symptoms of dementia (BPSD) . STELLA (Support via TEchnology: Living and Learning with Advancing AD) is a telehealth-based intervention that provides a personalized approach to teach care partners (CPs) strategies to manage BPSD. To understand the relationship between BPSDs and costs, weekly surveys were administered to CPs (n=12). Surveys asked about out-of-pocket costs incurred from: hospitalizations and emergency department (ED) utilization, primary care visits, use of paid in-home care, prescription drugs, and over-the-counter medications. The most frequent cost reported by CPs was prescription drug related (11 CPs), while costs associated with hospitalizations and ED were the least frequently reported (4 CPs). A longitudinal weekly survey-based approach to quantify CP costs is a novel approach examine an intervention outcome (cost) that matters to families and policymakers.


Flaviviridae) are a group of positive-sense RNA viruses that include well-documented agents of human disease. Despite their importance and ubiquity, the timescale of flavivirid evolution is uncertain. An ancient origin, spanning millions of years, is supported by their presence in both vertebrates and invertebrates and by the identification of a flavivirus-derived endogenous viral element in the peach blossom jellyfish genome , implying that the flaviviruses arose early in the evolution of the Metazoa. To date, however, no exogenous flavivirid sequences have been identified in these hosts. To help resolve the antiquity of the Flaviviridae, we mined publicly available transcriptome data across the Metazoa. From this, we expanded the diversity within the family through the identification of 32 novel viral sequences and extended the host range of the pestiviruses to include amphibians, reptiles, and ray-finned fish. Through co-phylogenetic analysis we found cross-species transmission to be the predominate macroevolutionary event across the non-vectored flavivirid genera , including a cross-species transmission event between bats and rodents, although long-term virus–host co-divergence was still a regular occurrence . Notably, we discovered flavivirus-like sequences in basal metazoan species, including the first associated with Cnidaria. This sequence formed a basal lineage to the genus Flavivirus and was closer to arthropod and crustacean flaviviruses than those in the tamanavirus group, which includes a variety of invertebrate and vertebrate viruses. Combined, these data attest to an ancient origin of the flaviviruses, likely close to the emergence of the metazoans 750–800 million years ago.The flavivirids (family  These viruses include well-documented agents of human and livestock disease, including dengue virus, hepatitis C virus, yellow fever virus, Zika virus, and Bovine viral diarrhea virus 1. Reflecting their regular occurrence as pathogens, our understanding of flavivirid biology is necessarily skewed towards a subset of metazoan hosts, particularly those known to experience overt disease or act as reservoirs for these viruses, impeding our ability to understand the evolutionary history of this family. Currently available data suggest that all established genera, with the exception of the genus Flavivirus, are vertebrate-infecting viruses and do not require an arthropod vector for transmission (The flavivirids (family smission .Flavivirus can itself be divided into four groups defined by phylogenetic position and host range: the (i) mosquito-borne flaviviruses, (ii) tick-borne flaviviruses, (iii) insect-specific flaviviruses, and (iv) vertebrate-specific flaviviruses, also known as the ‘no known vector’ flaviviruses    points tears ago . The phyFlaviviridae, we used the Serratus RNA-dependent RNA polymerase (RdRp) search (https://www.serratus.io/explorer/rdrp) to mine the Sequence Read Archive (SRA) database for novel flavivirid sequences. To supplement this analysis, total RNA-sequencing data of the tunicate Botrylloides leachii was generated and screened to identify additional flavivirid sequences.Transcriptome mining is a proven method of virus discovery that leverages previous investment in metagenomics . To unde2.2.1Flaviviridae with a threshold score of ≥50 . The de novo transcriptome assemblies available at the National Center for Biotechnology Information (NCBI) Transcriptome Shotgun Assembly (TSA) Database (https://www.ncbi.nlm.nih.gov/genbank/tsa/) (as of June 2021) were also screened using the translated Basic Local Alignment Search Tool algorithm (TBLASTN) under default scoring parameters and the BLOSUM45 matrix. Amino acid sequences from representatives of the four Flaviviridae genera along with the related jingmenviruses were used as queries for the palmID and TSA database searches . Adapter removal and quality trimming were conducted using Trimmomatic (v0.38) with parameters SLIDINGWINDOW:4:5, LEADING:5, TRAILING:5, and MINLEN:25 (de novo into contigs using MEGAHIT (v1.2.9) (Flaviviridae protein database using Diamond BLASTx (v2.0.9) with an E-value threshold of 1 × 10−5  . The ass1 × 10−5 . To iden2.4https://www.ncbi.nlm.nih.gov/tools/vecscreen/) and removed. Contig abundances were calculated using the RNA-Seq by Expectation Maximization software (v1.3.0) (Sequence reads were mapped onto virus-like contigs using Bbmap (v37.98), and areas of heterogeneous coverage were manually checked using Geneious (v11.0.9) . Where p(v1.3.0) . GetORF (v1.3.0) . To anno(v1.3.0) . Genome (v1.3.0) .2.5E-value cutoff of 1 × 10–20. In addition, the virus-like sequences were checked for host gene contamination using the contamination function implemented in CheckV (v0.8.1)  and a subset of the whole-genome shotgun contig database (as of October 2022) using the TBLASTN algorithm with an (v0.8.1) . All EVE2.6https://www.adobe.com)  .Flaviviridae genera (e.g. Harrimaniidae flavivirus). Virus–host assignments were made using a combination of host/virus abundance measurements and phylogenetic analyses. Where <80 per cent of host abundance was associated with the target species of the library, the possibility of alternative hosts was considered. In this case, the other organisms comprising this library were examined to determine if they might represent the source of the virus sequence. For instance, given the known host range of the flavivirids, it is more likely that these sequences are derived from metazoan species than from bacteria, fungi, or archaea. As such, metazoan species were given greater weighting when assigning putative virus–host assignments. Where host assignment proved difficult to assign with accuracy, the suffix ‘associated’ was added to the host name to signify this . Where the taxonomic position of a virus was ambiguous, the suffix ‘-like’ was used (e.g. African cichlid flavi-like virus).The virus sequences identified in this study were named using a combination of the host common name—if known—and the appropriate 2.7https://www.adobe.com)  employing the generalized affine gap cost algorithm . Poorly obe.com) .2.8Flaviviridae, we analysed the co-phylogenetic relationship between viruses and their hosts. Host cladograms were created using the phyloT software, a phylogenetic tree generator based on NCBI taxonomy (http://phylot.biobyte.de/). Virus–host associations were obtained from the NCBI virus database , flavivirus-like (7), pesti-like (4), and unclassified flavivirial-like sequences (1) , Fig. 1.3.1We identified seven putative flavi-like virus sequences, including cnidaria flavivirus (CnidFV) and cigar comb jelly flavi-like virus (CcjeFV) in libraries of the early diverging metazoan phyla Cnidaria and Ctenophora, harrimaniidae flavivirus (HarFV) in an acorn worm (Enteropneusta), photeros flavivirus (PhoFV) and sea-firefly flavivirus (SefiFV) in marine ostracods, Chowder Bay tunicate–associated flavivirus in tunicates (CbtuFV), and African cichlid flavivirus (AfciFV) in a cichlid fish . For allA range of genome structures was observed and found to be largely consistent with those found in this genus. For example, PhoFV and SefiFV, like the other viruses identified in marine crustaceans, are predicted to contain a programmed −1 ribosomal frameshift on a ‘slippery’ heptanucleotide sequence downstream of the NS1 region  Fig. 2,. HoweverFlavivirus along with the crustacean flaviviruses  within a larger diversity of marine crustacean flaviviruses. Two sequences, CnidFV and HarFV, fell basal to all classified members of the genus iviruses . Notablyiviruses . CcjeFV 3.2We identified four pesti-like virus sequences in amphibians, reptiles, and bony fish . Two ful3.3n = 6) add to the known diversity of bat, avian, primate, rodent, and treeshrew hepaciviruses , expanding on the two hepaciviruses previously identified in this group . The remiviruses . Of the iviruses . Partialiviruses . Of noteiviruses  and shar3.4D. oweni, a species of parasitic protist belonging to the phylum Apicomplexa. Two contigs were assembled from this library, 3689 and 4577 bp in length and predicted to contain the NS3 and NS5 domains, respectively  in ectively . DiowV sectively .3.5E. flexuosa (family Plexauridae) library in which CnidFV was assembled, a genus of unicellular microalgae, Symbiodinium (phylum Dinoflagellata), represented 64 per cent of all contigs  was assembled, although it is uncertain to which family of catfish this sample belonged. Likewise, the American bullfrog (Lithobates catesbeianus) transcriptome comprised 60 per cent contigs associated with fork-tongued frogs (Dicroglossidae) and 17 per cent associated with true frogs (Ranidae), including L. catesbeianus. No host-associated contigs were detected in the D. oweni library in which DiowV was assembled. Instead, 64 per cent of the library is composed of contigs associated with marine gastropod molluscs.Contigs belonging to catfish (order Siluriformes) comprised 95 per cent of the 3.6Flaviviridae, we estimated co-phylogenies to quantify the evolutionary relationship between the ‘pegi-hepaci’ and pestivirus clades and their hosts (Flavivirus were excluded because of the high frequency of vector-borne viruses). In accordance with earlier studies  among the  studies , this an studies . Two vir studies . GmebHV  studies . FrbuHV,Importantly, despite the widespread occurrence of cross-species transmission, virus–host co-divergence was also predicted to have occurred relatively frequently across the ‘pegi-hepaci’ and pestivirus clades, representing 22 and 23 per cent of all events, respectively. For these groups, duplication events were more uncommon, representing 10 and 6 per cent of total events . Extinct4.Flaviviridae and Metazoa and highlights the vast number of viruses that remain undiscovered.Through transcriptome mining, we identified 32 novel flavivirid sequences across the Metazoa, including the first flavivirus-like sequences in non-bilaterians, pestivirus-like sequences in amphibians, reptiles, and bony fish, as well as a range of vertebrate hepaciviruses. Hence, this work provides further evidence of the long-term associations between the Flavivirus than are the tamana/flavi-like viruses, suggesting that these groups, including the jingmenviruses, are evolutionarily distinct , we suggest that the origins of this group of viruses likely extend much further back in time than previous estimates and are closer to the emergence of the metazoans 750–800 million years ago . This codistinct . As suchronments .E. flexuosa in library SRR12876665, we have assigned the resulting virus sequence as cnidaria flavivirus (CnidFV). The high abundance of Symbiodinium in this library is unsurprising given that the octocoral-Symbiodinium mutualism is well known   . Co-infedis (Bd) . If Bd aWe identified 20 novel hepacivirus sequences, among which a clade of cichlid-associated hepacivirus sequences is notable. This clade was derived from a study of Lake Tanganyika, a freshwater lake shared by Tanzania, the Democratic Republic of the Congo, Burundi, and Zambia that is known for its high diversity of endemic cichlid species . ImportaBats and rodents harbour a large diversity of hepaciviruses and are thought to have played an important role in their global spread and broader evolutionary history . We idenIn very broad terms, we find that the hepaci-, pesti-, and pegiviruses cluster with the phylogeny of their hosts, with the relevant frequent cross-species virus transmission events only occurring within host classes . The exceptions were FrbuHV, Western African lungfish hepacivirus, and Wenling moray eel hepacivirus that fell basal to those viruses identified in cartilaginous fish  forms a sister group to the ‘pegi-hepaci’ lineage, although this may be artefactual due to recombination or extreme rate variation . If the Another notable observation from this study was the identification of a flavivirus in non-bilaterians, which raises additional questions on the ancestral mode of flavivirus transmission. Non-bilaterians lack the circulatory system of vertebrates, suggesting that an alternative mode of cell-to-cell virus transmission may exist in these animals .Flaviviridae and the Metazoa as a whole.In sum, through a broad-scale survey of publicly available and newly generated transcriptome data, we revealed a wide diversity of flavivirid sequences in undersampled metazoan species. In doing so, we provide additional information for an ancient origin of the flaviviruses, likely closer to the emergence of the metazoans some 750–800 million years ago, and hence for the long-term association between the veac124_SuppClick here for additional data file.


The coincidence of the monkeypox outbreak with the coronavirus disease 2019 (COVID-19) pandemic raises a global concern about a potential new pandemic and the possible consequences. As the World Health Organization declared the international monkeypox outbreak a global emergency, there is apprehension about the complications and mortality of monkeypox infection.The monkeypox virus is a zoonosis: a disease that is transmitted from animals to humans. It is an enveloped double-stranded DNA virus that belongs to the Orthopoxvirus genus. Human-to-human transmission occurs with close contact with respiratory secretions, sores on an infected person's skin, or contaminated items like clothing.Monkeypox is endemic in regions of Africa, however, because of smallpox eradication and a decrease in vaccination efforts, this led to an outbreak in the United States of America in 2003 and a new world outbreak in 2022.Most patients experience prodromal sickness with fever, malaise, and enlarged lymph nodes before developing a rash. In addition to skin lesions, individuals may also experience secondary skin and/or soft tissue infection, pneumonitis, ocular problems, and encephalitis. There is an increased risk of infection among men who have sex with men and human immunodeficiency virus infection (HIV) patients. The polymerase chain reaction is the gold standard for diagnosis. Management is usually supportive but some cases may require tecovirimat. This is a comprehensive review of monkeypox virus epidemiology, clinical features, and the most up-to-date, effective management and prevention. The monkeypox virus causes a smallpox-like illness in humans. It is an enveloped double-stranded DNA virus that belongs to the Orthopoxvirus genus . RabbitsMonkeypox is a zoonosis: a disease that is transmitted from animals to humans. When a person comes into contact with an infected animal, monkeypox can be transmitted to them. Avoiding unprotected contact with wild animals, especially those that are sick or dead, can lower the chances of contracting monkeypox from them (including their meat and blood) . Any iteClose contact with respiratory secretions, sores on an infected person's skin, or contaminated items like clothing and bedding can lead to human-to-human transmission. Although, extended face-to-face contact is often necessary for transmission by respiratory droplet particles. This puts close family members, household members, and other contacts of active patients at increased risk .A monkeypox epidemic was verified in the United Kingdom (UK) on May 6, 2022. It was brought about by a British citizen who visited Nigeria, where the disease is widespread. On May 4, this traveler returned to the UK, bringing the outbreak's index case with them .On Saturday 23/07/2022, the World Health Organization declared the international monkeypox outbreak a global emergency, the highest level of alert the WHO can issue . This deEpidemiologyThe first incidence of the monkeypox virus was the outbreak of a pox-like disease in 1959 in monkeys hosted at a research center in Copenhagen, Denmark . As the As of May 21, 2022, 92 cases have been verified globally, coming from 13 nations  where the monkeypox virus is not endemic .A total of 3413 laboratory-confirmed cases and one death have been reported to WHO from 50 countries and territories in five WHO regions since January 1 up to June 22, 2022. Two thousand thirty-three out of 3413, or 86%, of the laboratory-verified cases, came from the WHO European Region. The African Region , the Region of the Americas , the Eastern Mediterranean Region 15/3413, 1%), and the Western Pacific Region  are the other regions reporting cases. In the second quarter of 2022, Nigeria recorded one death [/3413, 1%Clinical signsMost patients experience prodromal sickness with fever, malaise, and enlarged lymph nodes following a 10-14 day incubation period before developing a rash -17. ClinTypically, the prodromal phase lasts one to three days before the classic maculopapular rash appears, The clinical progression is remarkably similar to that of typical smallpox lesions, and the mean diameter of the skin lesions is between 0.5 and 1 cm . LesionsIn addition to skin lesions, individuals may also experience extracutaneous symptoms such as secondary skin and/or soft tissue infection, pneumonitis, ocular problems, and encephalitis . The 10%DiagnosisMonkeypox can be diagnosed using the clinical signs, but to distinguish this illness from those caused by other potential etiologies, laboratory confirmation is required . GeneticUnfortunately, a lot of these techniques are quite general and cannot distinguish between infection with the monkeypox virus and infection with other poxviruses. For instance, histologically, the lesions caused by monkeypox are comparable to those caused by other viral exanthems  and feature ballooning keratinocyte degeneration, prominent spongiosis, dermal edema, and acute inflammation . OrthopoImmunoglobulin M (IgM) is thought to be more useful in diagnosing recent infections, even while IgG alone cannot offer a conclusive diagnosis to a patient who has been exposed to the orthopoxvirus throughout his life through vaccination .ManagementThe clinical course of monkeypox infection, fortunately, is often mild and self-limiting. Because of this, particular therapy is rarely warranted, and treatment is frequently supportive. Antipyretics for fever, analgesics for pain, and other forms of supportive therapy .The first antiviral medication to enter clinical use was methisazone, but it is no longer in use since it was quite hazardous when given systemically . A broadPreventionThe final consequence of the smallpox eradication campaign was the complete elimination of the disease worldwide. Monkeypox cannot be eradicated, regrettably, because a reservoir of animals is present . But inoPeople who are at risk should get vaccinated, according to certain nations. Smallpox vaccines may also be helpful for monkeypox. Only those who are at risk, for instance, someone who has had intimate contact with someone who has monkeypox, should be given the vaccine. At this time, widespread immunization is not advised . The EurIn any hospitalized patient with a broad vesicular rash of uncertain cause in whom monkeypox and smallpox are included in the differential diagnosis, standard, contact, droplet, and airborne precautions should be undertaken. Close contacts should also be kept an eye on . In the The introduction of a disease like monkeypox into a new, previously disease-free region of the world can provoke a global concern of potential pandemic. Social distancing and contact tracing are fundamental because human-to-human transmission most commonly occurs via respiratory droplets or direct contact with an infected individual.Given the sharp rise in positive cases, monkeypox's public health implications testing should be made more widely available, and awareness of the sickness caused by human monkeypox should also rise. The necessity for self-quarantine and the mechanisms of transmission must be explained to patients who are suspected or have been diagnosed. Increasing public awareness about the mode of transmission, clinical symptoms, and prevention methods will limit the progression of this disease. Recommendation for vaccination especially among high-risk individuals (men who have sex with men and HIV patients) will prevent the evolution of disease throughout the world. Monkeypox's early clinical manifestations may resemble those of other sexually transmitted infections, including syphilis, herpes, or lymphogranuloma venereum. In patients with epidemiologic risk factors for monkeypox, a thorough history should be conducted together with a physical examination. Appropriate specimens should also be gathered from lesions and tested for the monkeypox virus.


Though instances of arthropod-borne (arbo)virus co-infection have been documented clinically, the overall incidence of arbovirus co-infection and its drivers are not well understood. Now that dengue, Zika and chikungunya viruses are all in circulation across tropical and subtropical regions of the Americas, it is important to understand the environmental and biological conditions that make co-infections more likely to occur. To understand this, we developed a mathematical model of co-circulation of two arboviruses, with transmission parameters approximating dengue, Zika and/or chikungunya viruses, and co-infection possible in both humans and mosquitoes. We examined the influence of seasonal timing of arbovirus co-circulation on the extent of co-infection. By undertaking a sensitivity analysis of this model, we examined how biological factors interact with seasonality to determine arbovirus co-infection transmission and prevalence. We found that temporal synchrony of the co-infecting viruses and average temperature were the most influential drivers of co-infection incidence. Our model highlights the synergistic effect of co-transmission from mosquitoes, which leads to more than double the number of co-infections than would be expected in a scenario without co-transmission. Our results suggest that appreciable numbers of co-infections are unlikely to occur except in tropical climates when the viruses co-occur in time and space. Hence, the diseases' spatio-temporal distribution is largely determined by the same environmental and climatological drivers [The past decade has seen the Americas affected by epidemics of both Zika and chikungunya, adding to the burden of arthropod-borne (arbo)viral disease in a region where seasonal dengue epidemics were already a regular occurrence in most countries –3. All t drivers –7. This  drivers . The rat drivers .The phenomenon of arbovirus co-infection is still largely understudied with many unknowns . For insSeasonal climate drivers play an important role in arbovirus infection dynamics, as variations in temperature determine environmental suitability for mosquito vector survival and virus transmission –21. ArboData on the frequency of arbovirus co-infection remain sparse , and whe. 2. 2.1et al. [et al. [We used a deterministic SEIR-SEI model to explore the influence of temperature on arbovirus co-infection magnitude and timing. This model incorporates two arboviruses, referred to as virus A and virus B, with identical transmission and human recovery rate parameters. We relied upon several of the structural assumptions and parameter values reported by Vogels et al. , particuet al. ,24. Cons [et al. , we assu [et al. . VirusesWe additionally made several assumptions about the roles that exposed and infected individuals play in transmission. Co-infection in humans and mosquitoes can occur via co-transmission to susceptible individuals or sequential transmission involving individuals who are susceptible and then exposed to the first of two viruses. Following their infectious period, individuals recover, and if they have only been singly infected by one virus can then be singly infected with the other virus. Infection while an individual is exposed restarts the incubation period. In our baseline analysis, we assumed no cross-protection or enhancement from a prior infection but explored this in later analyses. Structural details of the model are illustrated in Aedes aegypti life history: mosquito biting rate, probability that an infected mosquito transmits to a human during feeding, probability that a mosquito becomes infected after feeding on an infected human, mosquito mortality rate and virus extrinsic incubation rate. Their values were chosen with reference to previous modelling work on fitted thermal responses for Aedes aegypti [We incorporated a seasonal component into the model by using temperature-dependent parameters, where biologically appropriate . We used aegypti . This ap. 2.1.1Model equations for humans are as follows, with parameter values and meanings shown in tables Model equations for mosquitoes are as follows, with parameter values and meanings shown in tables et al. [Tmax, Tmin and Tmean represent the maximum, minimum and mean temperature for a region across a calendar year. In equations (2.33) and (2.34), c, Tmax, Tmin and T represent the fitted rate constant, critical temperature maximum, critical temperature minimum and temperature at a given time, respectively. As in Mordecai et al. [Equations to address seasonal fluctuations in temperature and thermal traits across a 365-day period are as follows, modelled after the approach to seasonal forcing used by Huber et al. . In equai et al. , we assu. 2.1.2Aedes aegypti life traits and arbovirus transmission are shown in Fitted parameters describing m(T), remained biologically feasible regardless of climate, we followed the approach of Siraj et al. [γ, such thatγ is defined here as the product of the estimated ratio of mosquitoes to humans in Rio de Janeiro in 2012 (1.24) and the temperature-varying fitted mosquito mortality rate at temperatures typical of that city, such that TTropical = 24.3 (equation (2.35)) [m across various temperature environments ensured that the ratio of mosquitoes to humans remained biologically feasible, between 1.18 and 1.25 over the course of each simulation (equation (2.36)).Additional population-level parameters were governed by temperature. To ensure that the ratio of mosquitoes to humans, j et al.  and deve (2.35)) ,32,33. Uet al. [Aedes aegypti mosquitoes follow those used in previous dengue modelling studies [et al. [Temperature-independent parameter values and definitions are consistent with those in Vogels et al. , which d studies ,28,29 wh. 2.2. 2.2.1We focused on four model outputs: (i) cumulative incidence of infection with virus A, (ii) cumulative incidence of infection with virus B, (iii) cumulative incidence of co-infection, and (iv) proportion of all infections that were co-infections. All quantities were defined as cumulative values across the course of a year-long simulation, at which time all arbovirus outbreaks had run their course.. 2.2.2α), recovery time (r) and human incubation period (εH). We considered the individual impact of these parameters on model outputs in a series of one-at-a-time sensitivity analyses. For each parameter, we varied the value across a plausible range . These included an environment with temperatures typical of a tropical region  and an environment with temperatures typical of a more temperate region . Using monthly mean temperatures for the two cities, we calculated associated mean, minimum and maximum temperatures across a year to determine the average temperature and seasonality observed in the most recent full calendar year. We systematically considered each possible combination of virus arrival times within a simulation and compared simulation results between the two temperature settings. We performed additional simulations exploring the timing and magnitude of co-infections observed when the invading arboviruses are associated with asymmetric extrinsic incubation periods and variable incubation periods for co-infections. For these simulations, we assumed a CHIKV-like incubation period of 2 days for one virus and a ZIKV-like incubation period of 14 days for the other. We explored a range of assumptions around the incubation period of co-infected mosquitoes, using the 2 and 14 days as the upper and lower bounds, respectively.. 2.2.4To evaluate the interaction components of our model parameters, we conducted a global, variance-based sensitivity analysis, also known as a Sobol sensitivity analysis, using the SALib library in Python . This ap. 3. 3.1a,b). Simultaneous arrival of viruses A and B resulted in co-infection incidence ranging from 32 to 83 per 1000 individuals per year, with low incidence being associated with periods of significant negative temperature change, particularly in late summer (days 100–150) (a). Asynchronous virus arrival resulted in fewer co-infections than simultaneous arrival did, with larger gaps between virus arrival dates corresponding to lower incidence of co-infection (b).To better understand what combinations of epidemic timing and seasonality lead to a high incidence of co-infections, we explored a range of virus arrival times throughout the year under tropical and temperate temperature regimes. This scenario is reflective of a situation where two arboviruses are imported to a location within a year of each other, as happened with Zika and chikungunya viruses in some South American countries in the 2014–2016 period. When seasonal patterns resembled those in a tropical region , simultaneous (same-day) virus importation resulted in incidence of co-infection that was always greater than 19 per 1000 individuals, although seasonal differences were observed a,b. Simu100–150) a. Asynchnfection b.Figurec,d). Asynchronous arbovirus arrival resulted in seasonal trends in co-infection consistent with simultaneous arrival but produced negligible co-infections as the time between arrivals grew (d). Arrivals after the summer (around day 100) generated similarly negligible co-infection incidence, as temperatures fell below those conducive to virus transmission by Aedes aegypti mosquitoes (c).By contrast, when seasonal patterns resembled those of a more temperate region , both simultaneous virus arrival and seasonal high temperatures were required to observe non-negligible co-infection, with a maximum incidence of 0.0079 per 1000 individuals c,d. Asynals grew d. Arrivasquitoes c.These trends generally held true when the arboviruses were not assumed to be identical and dengue-like, and instead were modified to approximate the invasion of chikungunya and Zika viruses into a tropical environment. In the presence of asymmetry in the viruses’ extrinsic incubation periods, a phenomenon that has been documented clinically , we obseα) refers to the level of protection each infection provides against the other, where 0 indicates no protection, and 1 indicates complete protection. When values of α increased, the incidence of virus A decreased slightly, and the proportion of co-infections decreased dramatically . This decrease in the incidence of virus A when cross-protection was high was driven by individuals with virus B who experienced a reduced force of infection of virus A and was limited by the later arrival of virus B into the population. The proportion of co-infections declined steeply as high values of α inhibit both sequential and co-transmitted co-infections from occurring. However, high α favours co-transmitted co-infections over sequentially transmitted co-infections because co-transmission is limited by cross-protection to the extent that sequential transmission is.In addition to being temperature-driven, model outputs are influenced by several biologically important temperature-independent parameters, including immunological cross-protection, recovery time and the human incubation period. The coefficient of cross-protection , the average time in days it takes for a human to recover from either infection. Longer recovery times correspond to higher virus A incidence and a higher proportion of co-infections, as more time spent infectious allows for greater exposure to a second infection and increases the reproduction number of both viruses . Longer recovery times lead to a lower ratio of co-transmitted co-infections to sequentially transmitted co-infections for the same reason—time spent infectious, where another infection cannot be acquired immediately, favours sequential infection transmission (h).Model outputs are also noticeably influenced by changes in the value of recovery time  is also measured in days and has been approximated in studies of dengue and Zika virus to be 5–8 days [εH does not strongly influence the incidence of either single or co-infections . However, the seasonal and non-seasonal models produce noticeably different outputs depending on the day virus A is imported . When virus A was imported in fall or winter, the incidence of virus A and the proportion of infections that were co-infections were relatively low as compared with spring or summer importation days. While both the seasonal and non-seasonal models have parameters that approximate values for a tropical-like region, earlier importation dates within the year appear more suitable for arbovirus infection and co-infection, as rising temperatures following importation reach values ideal for transmission once incidence has grown.By contrast with the previous two parameters, model outputs do not appear particularly susceptible to changing incubation period values. Human incubation period  none, (2) 50% immune to virus A, (3) 50% immune to virus B, and (4) 50% immune to both viruses  . Scenaria,d). This resulted in a decrease in virus A incidence driven by the lower proportion of individuals not immune to virus B and, thereby, partially immune to virus A. By contrast, scenarios with immunity to virus A and immunity to both produced negligible incidence of virus A infections , since both limited the population susceptible to virus A.When high levels of cross-protection were present, immunity to virus B limited infection by either virus once virus B became prevalent a,d. Thisfections a,d, sincj). When there was no initial immunity, there was a larger group of individuals susceptible to virus A at the beginning, which provided more opportunities for sequentially transmitted co-infections. Since that population of individuals was much smaller when there was immunity to both viruses at the beginning, sequential transmission occurred less frequently. More generally, though, immunity of any type led to a much smaller proportion of infections that were co-infections (d–f). While this proportion was low even under the no-immunity scenario, immunity to even a single virus limited the occurrence of both sequential and co-transmitted co-infections.Pre-existing immunity had the most noticeable impact on the ratio of co-transmitted co-infections to sequentially transmitted co-infections, where we observed that co-transmitted co-infections were more heavily represented under the scenario with immunity to both viruses (black line) than they were under the no-immunity scenario (red dashed line) j. When tfections d–f. Whil. 3.3To gain a holistic view of the effect of the examined biological and environmental factors and their interactions on the epidemiology of arbovirus co-infection, we used a variance-based sensitivity analysis. This approach allowed us to explore the contribution of each parameter to the variance of each model output. Examining the first-order indices, or those describing the direct relationship between each parameter and each model output, revealed that interactions between parameters accounted for more than 50% of the total variance in all four outputs . HoweverWe further explored the interactions between parameters that contribute to variance in the incidence of co-infection and found these relationships to be consistent with the results from first- and total-order indices alone . InteracAdditionally, we assessed the role that interactions between the viruses themselves play in observed co-infections, particularly via the mechanism of co-transmission. As our model allows for simultaneous arbovirus co-transmission between mosquitoes and humans, a phenomenon documented frequently in laboratory studies , the fre. 4By incorporating seasonal temperature variation, differential importation times and virus co-transmission, the model implemented here explores the drivers of arbovirus co-infection and assesses conditions under which increased arbovirus co-infection may be likely. To observe substantial co-infection incidence, our model suggests a need for both the consistently favourable temperatures typical of the tropics as well as temporal synchrony between the viruses. In more temperate regions co-infections of the studied arboviruses are rare, only occurring during summer months and even then at very low levels. Repeated seasonal arbovirus outbreaks could result in some degree of cross-protective natural immunity in populations living in tropical environments ,38,39, wAedes-borne disease incidence and higher poverty levels [This study provides a novel exploration of temperature variation in mosquito life traits and co-infection dynamics within a single modelling framework. Previous modelling studies have characterized the relationship between temperature and arbovirus transmission and have emphasized that warm climates and highly variable moderate climates have high epidemic suitability ,40. Our y levels  indicateWe made several assumptions to support parsimonious and computationally tractable model scenarios. First, while temporal synchrony between arriving arboviruses was a crucial component of this study, it is perhaps more likely that an arbovirus would be introduced to an environment where another arbovirus is already endemic, as has been noted in studies of Zika virus and endemic dengue in the Americas ,41. We eR0), as well [Second, for the sake of simplicity, we assumed that the population was homogeneous and well-mixed. This could lead to an overestimation of transmission, as high-risk clusters are not always localized both in time and space . Incorpo as well ,44. WithThird, by primarily modelling the arboviruses as identical and dengue-like, we have not considered some features specific to ZIKV, CHIKV or other arboviruses. Experimental studies characterizing arbovirus extrinsic incubation periods have reported enhancement effects in sequential CHIKV–ZIKV co-infections via a shortened ZIKV incubation period , as wellγ(T), a parameter used to ensure that mosquito populations remained within a reasonable range. While this neglects many complexities of mosquito population dynamics, our use of simplifying assumptions more generally made it possible to isolate the effects of parameters of interest in a straightforward way. Sensitivity analysis of our model allowed us to further examine all parameter interactions and explore the full parameter space.Finally, model parsimony also influenced our choice of a single mosquito population scaling factor, In this study, we built upon existing modelling work  to explo


It combines elemental iodine (I2) and fulvic acid (C14H12O8), forming a clathrate compound. The antiviral properties of Iodine-V reduce viral load in the air to inhibit viral transmission indoors. This antiviral property was applied to form a disinfectant solution called SAFEAIR-X Aerosol. The current study evaluates the antiviral efficacy of Iodine-V in aerosol form in a prototype called SAFEAIR-X Aerosol. The experiment measured the antiviral efficacy of SAFEAIR-X following exposure to the Vaccinia virus (VACV) samples as a confirmed surrogate for SARS-CoV-2. The SAFEAIR-X showed 96% effectiveness, with 2 seconds of spraying duration and 60 seconds of contact time releasing less than 0.0001 ppm of iodine into the air, and a log reduction value of 1.50 at 60 seconds in 2 out of 3 tests was observed. Therefore, this study demonstrates SAFEAIR-X aerosol as a potential indoor surface and air disinfectant.Iodine-V ((C The novel coronavirus known as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first detected in December 2019 in Wuhan, China which manifests severe symptoms like difficulty in breathing and often results in the death of the affected person . The smaThe viral load in the air gradually reduces because the heavier and larger respiratory droplets fall to the ground due to gravity, and the very fine droplets mix with the airstream , 9. In avia respiratory droplets and condensed viruses that exist for some time on surfaces [SARS-CoV-2 is transmitted person-to-person surfaces , 14. Howsurfaces , 16. It surfaces , 18.The primary mechanisms for disseminating the SARS-CoV-2 virus are close physical contact and the shedding of viral particles from mucous membranes .An ideal strategy for preventing viral transmission is decreasing the respiratory droplets in the air, in addition to the mechanical or natural ventilation that also reduces the viral load of SARS-CoV-2 . Two air26H39N4O15)x * (I2)y) is an aqueous solution that is composed of water-soluble elemental iodine (I2) and fulvic acid (C14H12O8) to form a clathrate compound. It forms a stable complex inert to nitrogen and oxygen in the air [in vitro, and 99% efficacy was observed against SARS-CoV-2 in 3 dilutions [The current study applies Iodine-V as an aerosol called SAFEAIR-X, which is a prototype of air and surface disinfectant, to evaluate its efficacy against airborne viruses. Iodine-V (formula (C the air . In addi the air . EIDs ar the air  have shoilutions . The iod2) has broad-spectrum antimicrobial activity, and povidone-iodine (PVP-I) has been tested and applied for decades as an ideal antiseptic [2 forms a complex with the polymer PVP (synthetic carrier), which has no antimicrobial activity. But Iodine-V is composed of I2 and fulvic acid (C14H12O8), a synthetic carrier and novel antimicrobial molecule [2 in PVP-I is a potent microbicidal agent with 99.99% virucidal efficacy at a concentration of only 0.23%, regardless of the virus type. Mechanism of action involves iodine rapidly penetrating microorganisms and important oxidizing nucleotides, fatty acids, and proteins causing cell death [in vitro study against SARS-CoV-2 [Elemental iodine  with MERS-CoV, SARS-CoV-1, and EBOV using PVP-I, which showed VACV to be the most resilient virus [The present research tested SAFEAIR-X Aerosol against the Vaccinia virus (VACV) as a surrogate of SARS-CoV-2. The selection of surrogate viruses is crucial when creating a standardized test. High resistance to drying and disinfectants, uncomplicated viral multiplication in cell culture, availability in virus collections, and a low BSL are prerequisites for conventional surrogate viruses (BSL 1 and 2) . The VACnt virus , 28. WheThe objective was to develop an aerosol in a device for domestic, pharmaceutical, veterinary, and commercial indoors use.In this experiment, SAFEAIR-X was tested against VACV as a surrogate of SARS-CoV-2. When evaluating the virucidal efficacy of alcoholic formulations for disinfectants, VACV is found to be safe and an apt substitute –31. The The experimental control used was according to standard EN 17272. The research aimed to examine the processes involved in dispersing a chemical disinfectant in gaseous, steam, or aerosol form. The test compared the number of microorganisms that survived the applied air disinfection process and analyzed the reduction in test microbes detected on nonporous surfaces . The micThe experiment was conducted over 14 days, from August 12 to August 27, 2021, taking the Vaccinia virus and Vero cells as the experimental specimens. The experimental conditions are listed in 3 and were treated via airborne disinfection from a distance without any manual application. The experimental conditions include the height of the test carriers at 1.0 m, contact time (t) was 1 min ± 10 s, the applied test temperature was 20°C ± 1°C, and the incubated temperature was 37°C ± 1°C + 5% CO2. SAFEAIR-X in the room was emitted from a distance of 3.0 m and a height of 1.5 m, with a relative height of 0.5 m from the carriers. Cytotoxicity control, interference control, and neutralization control were used to validate the assays. The virus was neutralized by diluting in Eagle’s Minimal Essential Medium (MEM) with 5% v/v fetal bovine serum (FBS) at 4°C, followed by gel filtration. The discs were transferred to a tube containing 1 ml of MEM with glass beads and vortexed to remove any residual viral particles from the surfaces. This dilution was the first step of neutralization. Microspin gel filtration column was used to complete the process by taking a small portion of the above suspension.The Vaccinia virus was obtained and isolated in the laboratory in this experiment. The viral suspension was dried on a stainless steel disc and placed away at a defined distance . The ino3 room, which released less than 0.0001 ppm of iodine content; hence, SAFEAIR-X exposure to the individuals in the room was no more than 0.1 ppm, which was harmless.The virucidal activity of SAFEAIR-X was compared to a control. The inoculation and emission were given for 2 seconds, and contact with the Vaccinia virus VR-1549 Elstree strain was for 60 seconds, including the time of diffusion through the air of the confined room. Inhalation of less than 0.1 ppm concentration of iodine does not irritate the mucous membranes, but inhaling more than 0.1 ppm of iodine is lethal . SAFEAIRThe experiment was performed thrice, and the samples of viral suspension were labeled as R1, R2, and R3.50). TCID50 assay describes the virus dilution amount that can induce cytopathic effects (CPE) in 50% of wells containing experimental inoculated cell culture after a defined period of time. A parallel experiment was also conducted with the Vaccinia virus ATCC VR‐1549/Vero cells. TCID50 was calculated by using the Kärber method [50 6.00).The experiment was set up for a basic virucidal efficacy test with three test replicates and a contact time of 60 seconds. Inoculation of the virus was done on a stainless-steel disc and allowed to dry. The carriers were placed from a defined distance away from the emission point and facing away from the emission direction. The carriers were exposed to the disinfectant by air diffusion without manual application. Neutralization was done with serial dilutions, and titered virus was transferred to 96‐well tissue culture plates to estimate the viability of the virus through the median tissue culture infectious dose-50 , so the more infectious virus recovered, the more host cells are killed. For the assay to work, the virus (and only the virus) is killing the host cells should be ascertained. In this control, we add the liquid media to the test sample, wait for 5 minutes, recover the media, and add it to the host cells. The virus has been removed, so if the host cells die, toxicity must be from the test sample. A result like this can invalidate the whole test. It should also be noted that this test only assesses the cytotoxicity toward cultured cells growing in media under specific lab conditions and is designed to support the antiviral test’s conclusions.The viral endpoint titration was exposed to 3 different sub-lethal concentrations of neutralized disinfectant and measured its effect on the pathogenicity of the virus concerning the titer achieved for control cells. This was conducted to evaluate the susceptibility of host cells to infection when a sub-lethal disinfectant concentration is applied. A lower susceptibility of host cells to infection after being treated by the disinfectant could lead to an overestimation of the product’s efficiency as fewer cells were infected. A volume of the neutralized disinfectant and a volume of phosphate-buffered saline (PBS) was applied in parallel to the host cells. The virus was then inoculated into the host cells, and the pathogenicity difference was estimated.The highest disinfectant concentration was applied to the virus, and the mixture was instantly removed and neutralized. The neutralized viral titer was measured to evaluate the neutralization procedure.Two discs were inoculated in the laboratory under similar environmental conditions as an experiment with ‘Recovery Control—Start’ and ‘End’ controls and were allowed to dry. The experiment recovered one disc at exposure time (t = 0 minutes), ‘Recovery Control—Start’ before the beginning of the test, and the other disc, ‘Recovery Control—End,’ was left at room temperature for exposing the virus for control of degradation over time and then recovered. The viral recovery control was dried on a disc and treated with sterile water. This was done for 15 minutes and then recovered. This served a similar purpose as the standard 15-minute contact time start/end recovery control. These results indicated a stable condition for the virus under experimental conditions and were not significantly degraded.The virus was exposed to 0.7% W/V formaldehyde, and TCID50 assessed the viral recovery at 5 and 15 minutes to examine the retained reproducible disinfectant resistance when the test virus was added. In addition, the cytotoxicity of neutralized formaldehyde was measured to assess the sensitivity of the assay.The current study only used the Vaccinia virus VR‐1549 Elstree strain and excluded human participants. Hence, ethical approval and patient consent were not needed for this study.50/ml with standard deviations [The viral titers were validated using the Kärber method and expressed as TCIDviations , 36. Theviations , 38.The research aimed to identify the viral efficacy of SAFEAIR‐X with a 2-second emission time and a 60-second contact time. Hence, no other theoretical analysis was done, and no more test data were recorded.The effectiveness of SAFEAIR‐X as a disinfectant for indoor settings against VACV was observed. The SAFEAIR‐X Aerosol covered an enclosed area with fine dry droplets, and the device is stable in presence of water, dust, or other inorganic particles lingering in the air.The prototype SAFEAIR-X Aerosol samples showed considerable virucidal efficacy against VACV. The contact time was only 60 seconds in the experiment for observation, but, as per the protocol, the measures of viral control were taken at different time intervals. Tables 50 formula, a value of ‘0.00’ represents ‘3.16E+01’which indicates a lower limit calculation instead of the presence of the virus. The number of wells (96-well plate) positive for a cytopathic effect (CPE) was noted and converted to a proportion of total wells. The cytotoxicity level was calculated using the formula 10*10 (n+0.5) [(n+0.5) = 31.6. The log of this value is 1.50. This defines the lower limit of the calculation.According to the TCID (n+0.5) . The calThe cytotoxicity of the prototype solution had no effect on cell morphology, growth, or susceptibility to the virus in experimented mixture dilutions. SAFEAIR-X product did not show any cytotoxicity.10 for the virus.This procedure’s validation of neutralization is known as the disinfectant suppression test. Column chromatography through an Illustra Microspin S‐400 HR column was performed to neutralize the disinfectant to achieve the best validation results. There was effective neutralization of the virucidal activity of the disinfectant at a dilution concentration of 100.0% v/v, as the difference observed was not greater than 0.5 log10, which was less than 1.0 log10, suggesting no interference of cytopathic effects with the dilutions of the disinfectant to sub-acute levels of viral production. The disinfectant-treated and PBS-treated cells showed a difference of (-0.16 + -0.17) = -0.33 logIodine has broad-spectrum antimicrobial activity against bacterial, viral, fungal, and protozoan pathogens and has been used as an antiseptic to prevent infection and treat wounds for decades. Coronavirus transmission can be primarily influenced by humidity, but the temperature could still moderately regulate the spread of COVID-19 . Existinvia chemical disinfection. These are categorized as enveloped viruses, large unenveloped viruses, and small unenveloped viruses. SARS-CoV-2, monkeypox, and VACV are examples of enveloped viruses that are more sensitive to chemical disinfectants than their non-enveloped counterparts [in vivo and may be considered a nasal or oral antiseptic to reduce virus transmission [Based on past research, viruses are also classified according to the increasing difficulty of inactivating the viruses terparts . Iodine-terparts . Iodine-terparts . Iodine-smission .Though the origins of the broad-spectrum antiviral activities of PVP-I have not been fully studied, there is evidence showing that iodine inhibits virus attachment to the host’s cell surface by blocking its receptors. It can also inhibit the enzymatic activity of viruses such as neuraminidase which are necessary to release viruses from host cells that prevent the infection spread to other uninfected cells . Chemica50). This assay was validated by cytotoxicity, interference, and neutralization controls. 50/ml), which indicates that there were no cytotoxic effects of the SAFEAIR-X aerosol on VACV in the diluted samples; the starting viral titer remained as such. Also, the viral morphology and growth remained unchanged, demonstrating negative cytotoxic effects. 50/ml at the Recovery Control “End” and 1.00E + 05 TCID50/ml for virus recovery).SAFEAIR-X Aerosol prototype is analyzed for its virucidal efficacy on the VACV cultures grown in Vero cells, where VACV was taken as a surrogate for the SARS-CoV-2 along with a control. It was tested on surfaces in dried form. SAFEAIR‐X was tested under standardized, nearly real-world settings to observe the effectiveness of inactivating viruses at the lowest concentration of 0.0001 ppm. The experiments supported the aerosol’s performance in a lab setting and highlighted SAFEAIR‐X’s potential to be applied as a disinfectant. The aerosol was exposed to the confined room for only 60 seconds. Viral titers were calculated as per the Kärber method, and the virucidal efficacy was measured as the median tissue culture infectious dose (TCIDThe experiment gave reproducible results in three repeats which are demonstrated in 3 confined room. SAFEAIR-X can significantly reduce the viral load on surfaces in indoor settings within 60 seconds and theoretically in the air because halogen gases are highly efficacious against viruses in lower concentrations [The study aimed to confirm the use of Iodine-V, a component of air disinfectant for indoor settings . The protrations , 41. IodSAFEAIR-X is also safe; it releases only 0.0001 ppm of iodine into the room during the research, giving options for extending the spray time or increasing the aerosol concentration to enhance its efficacy further while still maintaining a safety limit under 0.1ppm.