Stretching and foam rolling are common warm-up exercises and can acutely increase the range of motion (ROM) of a joint. However, possible differences in the magnitude of change on ROM between these two interventions on the immediate and prolonged effects  are not yet well understood. Thus, the purpose of this review was to compare the immediate and prolonged effects of a single bout of foam rolling with a single bout of stretching on ROM in healthy participants.In total, 20 studies with overall 38 effect sizes were found to be eligible for a meta-analysis. For the main analysis, subgroup analysis, we applied a random-effect meta-analysis, mixed-effect model, respectively. The subgroup analyses included age groups, sex, and activity levels of the participants, as well as the tested muscles, the duration of the application, and the study design.P = 0.39) nor a difference 10 min , 15 min , and 20 min  post-intervention. Moreover, subgroup analyses revealed no other significant differences between the acute effects of stretching and foam rolling (P > 0.05).Meta-analyses revealed no significant differences between a single stretching and foam rolling exercise immediately after the interventions (ES = 0.079; If the goal is to increase the ROM acutely, both interventions can be considered as equally effective. Likely, similar mechanisms are responsible for the acute and prolonged ROM increases such as increased stretch tolerance or increased soft-tissue compliance. Stretching with its varying techniques   effects of foam rolling and stretching on ROM, the time course  of the changes in ROM following these modalities is highly relevant for sports practice . Whilst studies reported an increased ROM following stretching  between single sessions of foam rolling and stretching on ROM and to summarize the mechanisms underlying ROM increases in both foam rolling and stretching based on the existing literature. According to the existing evidence, it was hypothesized that foam rolling compared to static stretching applied over the same duration would produce comparable immediate and time course effects in the increase in ROM.Prolonged static stretching (>60 s per muscle group) in isolation (no aerobic or dynamic activities within the warm-up) has been reported to induce performance impairments  and the search period was until the 5th of November 2021. The keywords for the online search were the same for the various databases and were  AND (stretch*). The systematic and independent search was conducted by three researchers . All the hits were screened by their title and abstract. If the eligibility of a paper remained unclear, the full text was further screened. Following this independent search, the findings of the researchers were compared. Disagreements were resolved by jointly reassessing the studies against the eligibility criteria. Following the removal of 97 duplicates in total, 102 papers were screened, where finally 18 papers found to be eligible for the meta-analysis. Additionally, two further papers from researchers’ libraries  were found to be eligible. No further eligible papers were found following an additional search of the references (search through the reference list) and citations (search through Google Scholar) of the already included papers. Consequently, in total, 20 papers  or parallel group design (pre- to post-comparison). However, we excluded studies with a parallel group design where only post-intervention values were compared. Moreover, we excluded studies which investigated combined effects of foam rolling and stretching. We further excluded conference papers or theses.From all the included papers, the characteristics of the participants , the sample size number, the study design , the characteristics of the intervention , the muscles tested by the ROM test, and the pre- and post-intervention values plus standard deviation of the main variable ROM of both groups  were extracted. If the full paper did not provide all the data required for the meta-analysis, the corresponding authors were contacted via email and Research Gate.I2 statistics were calculated among the effect sizes, and thresholds of 25%, 50%, and 75% were defined as having a low, moderate, and high level of heterogeneity, respectively  software according to the suggestions of Borenstein et al. . ConsequTo assess the methodological quality of the included studies, the PEDro scale was used. Two independent researchers  assessed 11 methodological issues by assigning with either one or no point. Note that studies with a higher score represent a higher methodological quality. If any conflict between the ratings of the two researchers was found, the methodological issues were reassessed and discussed. Moreover, to assess a possible publication bias, the statistics of the Egger’s regression intercept test was used.In total, 20 studies investigated the immediate effects of both a single foam rolling exercise and a single stretching exercise on ROM, and overall, 38 effect sizes were included for this meta-analysis. Additionally, out of these 20 studies, the time course effects of 10, 15, and 20 min post-intervention were investigated by five, four, and three studies, respectively. In summary, 411 participants with a mean age of 23.4 (±4.9 years) participated in the included studies. Moreover, Table P = 0.38) but also for the time course effects 10 min post-intervention , 15 min post-intervention , and 20 min post-intervention  indicate that no reporting bias is likely.Egger’s regression intercept test for the immediate effects . Both assessors agreed with 95.9% of the overall 220 (20 studies × 11 criteria) criteria. However, the mismatches were discussed and the assessors finally agreed with the scores presented in Table P = 0.39; I2 = 60.18). Moreover, Fig. The average percentage increase in ROM in the included studies following stretching, foam rolling was 7.2 ± 8.7%, 7.2 ± 5.5%, respectively. The meta-analysis revealed no significant difference between the two modalities  − 0.101 to 0.259; Q = 0.35; P = 0.56), activity level of the participants  , tested muscle by the ROM test  , duration of the application  , sex  , stretching technique  , and the study design  .A summary of all the subgroup analyses is shown in Table P = 0.65; I2 = 0.00), − 0.011  − 0.266 to 0.244; P = 0.93; I2 = 0.00), − 0.161  − 0.451 to 0.128; P = 0.275; I2 = 0.00), respectively.The average percentage increase in ROM 10 min post-intervention in the included studies following stretching, foam rolling was 6.7 ± 3.6%, 7.6 ± 4.8%, respectively. Moreover, the average percentage increase in ROM 15 min post-intervention following stretching, foam rolling was 11.6 ± 7.0%, 10.5 ± 5.6%, respectively. Finally, the ROM 20 min post-intervention was 4.5 ± 3.7%, 5.9 ± 3.6%, for stretching, foam rolling, respectively. The meta-analyses on the time course revealed no significant difference between the modalities at 10 min, 15 min, 20 min post-intervention with an effect size of − 0.051  − 0.274 to 0.172; Moreover, Fig. P = 0.39) nor 10 min , 15 min , and 20 min  post-intervention. Moreover, subgroup analyses revealed no differences (P > 0.05) between the age groups of the participants , activity levels of the participants , tested muscle by the ROM test , duration of the application , sex , stretching technique , and the study design .The purpose of this review was to compare the immediate and time course effects of a single foam rolling and stretching exercise on ROM. The results revealed no significant difference between the two modalities immediately after the interventions  to 90 s (8 studies) of rolling with only two studies each intervening with 120 s or 180-s of rolling. It would be of interest to note whether shorter or longer durations provide similar increases in ROM. Furthermore, 5/20 studies tested ROM with the sit-and-reach test. The ROM measured in the sit-and-reach test can be attenuated by a restrictive lower back even if the rolling had favourable effects on the hamstrings extensibility. Hence, the rolling-induced effects on hamstrings extensibility with this type of hip flexion testing could have been underestimated. Only 3/20 studies monitored upper body ROM  and thus the extent of possible rolling-induced ROM differences may be influenced by anatomical location and should be further considered. As is typical of exercise or sport science research, the predominant mean age of the participants was between 20 and 27 years with only two studies each examining participants with an average age between 31–37 and 15–16 years respectively. Additional studies are suggested to examine youth at different stages of peak height velocity (pubescence) as well middle-aged and elderly populations. Furthermore, the vast majority of foam rolling-related studies report on the effects of rolling on ROM and performance but very few evaluate mechanisms , activity levels of the participants , tested muscle by the ROM test , duration of the application , sex , stretching technique , and the study design . Hence, if the goal is to increase the ROM acutely, both interventions can be considered equally effective. However, foam rolling rather than isolated static stretching without aerobic or dynamic activities within the warm-up should be implemented as a warm-up when ROM and performance  are equally important determinants for the subsequent training or competition. Likely, similar mechanisms are responsible for the acute and prolonged ROM increases such as increased stretch tolerance or soft-tissue compliance.


Orthopoxvirus genus of the Poxviridae family is known as the monkeypox virus. The Congo Basin  clade and the West African clade are the two clades of the monkeypox virus. The virus was first identified in monkeys in a Danish laboratory in 1958, hence it was named monkeypox [The biggest-ever outbreak of monkeypox disease in non-endemic countries started in May 2022 . The Woronkeypox . The fironkeypox . In certSo far, cases were found in the USA, Canada, Australia, Israel, and several European countries including the UK, Sweden, Spain, Portugal, Netherlands, Italy, Germany, Belgium, and France. Most cases of monkeypox occur in central and western Africa . Human iOn May 22, the Bangladeshi government announced a health alert amid the outbreak of the monkeypox virus and noted that no one has yet been infected with the monkeypox virus in Bangladesh. However, it is almost certain that the virus will spread throughout the country. A number of preventative actions must be taken to stop the COVID-19 pandemic from further decimating Bangladesh's already-struggling healthcare system . The COVIn order to address this issue in limited resources countries like Bangladesh, doctors must be sufficiently knowledgeable about the disease's presenting signs and symptoms to ensure that suspected patients are promptly quarantined rather than simply receiving symptomatic treatments. In order to stop the spread of the contagious virus, hospitals should also have well-equipped isolation units ready to place patients under quarantine right away. Additionally, effective surveillance systems must be created in order to track cases and respond appropriately. Public education sessions on the atypical pox rash and its progressive stages, which include atypical macules, papules, vesicles, pustules, and scabs, are among the preventive actions that can help in dealing with any suspected case before it spreads.Monkeypox has no known cure, but according to health professionals worldwide, the smallpox vaccine is 85% effective in preventing illness. However, epidemiological studies, transmission dynamics, and the ecology of the disease are still not fully understood and need further focus. The animal reservoir is yet unclear, and case management and effective outbreak response strategies are poorly documented. Up-to-date complete, consistent and longer-term research is sorely needed to inform and guide evidence-based response and management of monkeypox outbreaks.Not applicable.None.I, Yasmin Jahan have participated in the study concept or design, literature search, produced a table, and wrote the final manuscript of the paper.The author declares that there is no conflict of interest.1.Name of the registry: Not applicable2.Unique Identifying number or registration ID: Not applicable3.Hyperlink to your specific registration (must be publicly accessible and will be checked): Not applicableYasmin Jahan.Not commissioned, externally peer-reviewed.Not applicable.


The aim of this in vitro study was to investigate viruses’ stabilities on manual toothbrushes using feline coronavirus (FeCoV) as representative of coronaviruses and an Avian influenza A virus H1N1 for influenza viruses.Two viruses, FeCoV  and H1N1 , were used in this study. Manual toothbrushes were disassembled into bristles, bristle fixation, and back of the toothbrush head, contaminated with the viruses and air-dried for 24 h. In a second experiment, whole toothbrush heads were contaminated, rinsed with water (5 ml for 15 s) and then air-dried.6.41, back of head: 106.81 and bristles: 106.63 TCID50/ml. Following air-drying of 12 (fixation) and 24 h, titers of ≤ 102.5, 103.75, and 102.72 TCID50/ml were found in the respective groups, with a detection limit of 102.5 TCID50/ml. For H1N1, immediately after contamination, the following average titers could be recovered: fixation: 105.53, back of head: 105.97 and bristles: 105.75 TCID50/ml. Following air-drying of 8 (fixation) and 24 h, titers were ≤ 102.5, 103.63, and 103.53 TCID50/ml in the respective group, again with 102.5 TCID50/ml being the detection limit. In case of water rinse, no infectious virus could be recovered after 12 h.For FeCoV, immediately after contamination, the following average titers were recovered: fixation: 10Viral load of both viruses is reduced by air-drying, especially following water rinsing.Clinical relevanceThe toothbrush itself plays an insignificant role in the self-transmission of coronavirus and influenza virus. During the past 2 years, a viral pandemic with a novel human coronavirus  lead to a global public health crisis. This circumstance caused a high impact on daily life, including dentistry and oral hygiene issues . The virGenerally, the toothbrush has been examined as a potential source of microorganisms in different studies, including both bacterial and fungal species –8. WhileAccordingly, this in vitro study had two aims: (I) it was examined whether two different viruses of importance and known to be transmitted via respiratory aerosols, i.e., coronavirus and influenza virus, would be detectable in a clinically relevant load on different parts of the manual toothbrush, depending on the time of air-drying. In this study, the feline coronavirus and the avian influenza A virus H1N1 were used. (II) It was also tested, if rinsing with water could reduce the viral load on the manual toothbrush. For this, a common high-quality standard protocol for testing viral load was applied. To differentiate the findings, different areas of the toothbrush were examined. It was hypothesized that viral load would be detectable at all parts of the toothbrush, while both air-drying and water rinsing lead to elimination of the titer of both viruses.7.5 TCID50/ml. AIV H1N1 was propagated in chicken embryo fibroblast to a titer of 106.5 TCID50/ml. The cultivation of the viruses was conducted at 37 °C and 5% CO2. The experiments were performed separately with each of the two viruses to assess the respective characteristics of the viruses.In this study, FeCoV (strain Munich) was applied as a representative virus for SARS-CoV-2. Furthermore, an avian Influenza A Virus H1N1 (RE 230/90) virus was used as an analogue for a human influenza virus. FeCoV was propagated in Crandell Rees Feline Kidney (CRFK) cells to obtain a titer of 10Dr.BEST Original, CLASSIC; GSK Consumer Healthcare, D-80258 München, CH-6343 Rich) were bought from public shops and used as test material. They have standardized flat bristles and were selected in the hardness grade medium. For examination of the contamination, (I) toothbrushes were assembled inside the laminar flow cabinet class II under sterile conditions to the three parts: bristle fixation, the back of the toothbrush, and bristles, which have been investigated separately. (II) For the second experiment, the entire toothbrush head was used.Manual toothbrushes . For each time period, the proceeding was assessed with 8-fold repeats. The experimental flow is displayed in Fig. 10 steps. After 2 days, the cell plates were examined under an optical microscope to investigate the cells on a cytopathic effect as a verification for the existence of remaining virus. The cell observation was proceeded for about 7 days to determine the 50% tissue culture infectious doses (TCID50) according to the Spearman–Kaerber method.To recover the remaining virus on the toothbrush areas, 4950 μl of phosphate buffered saline (PBS) was added to each cup of the 6-well plate after each drying phase. The areas were washed 10 times with the PBS to ensure that the virus was completely suspended. Subsequently, 200 μl of each area were transferred to a 96-well-PBS dilution plate, and titrated in logThe experimental flow is shown in Fig. After the drying phase, 5 ml of PBS was added into the cup of the 6-well plate to rinse the toothbrush head and bring the remaining virus on the toothbrush in solution.200 μl of each cup  were transfused to a 96-well-PBS dilutional plate. The subsequent procedure was conducted as depicted above. 100 μl of each serial dilution was transfused to the appropriate cell culture system in each case.2. As described in the procedure above, the plates were examined under the microscope for 7 days to investigate the cells on a cytopathic effect as a positive verification for virus. By means of the results, the titer could be determined according to the Spearman–Kaerber method.For the FeCoV, Crandell Rees Feline Kidney (CRFK) cells were used; for IAV H1N1, chicken embryo fibroblasts (HEF) were utilized for cell culture. Thereafter, the plates were incubated at 37 °C and 5% of COAll experiments were performed with 8-fold repeats. For statistical analysis, the software GraphPad-PRISM was used.7.5 TCID50/ml. Immediately after contamination, a titer loss of approximately 1 log10 level was detected in all three contaminated parts of the toothbrush. This titer-reducing tendency continued throughout the drying period. After only 12 h, no infectious residual virus could be detected on the bristle fixation. Little residual infectious virus was still detected on the back of the brush as well as on the bristles after 24 h of drying with titers of 103.75 TCID50/ml and 102.72 TCID50/ml, respectively. In some repetitions, the smallest detectable titer of ≤ 102.5 TCID50/ml was determined for the bristles after 24 h of drying.The results of the first experiment displayed that the verified remaining viral burden on the toothbrush diminishes with increasing drying time and is dependent on the respective toothbrush part Fig. . The FeC6.5 TCID50/ml. Right after the contamination of the toothbrush areas, the titers of the remaining virus on all tested toothbrush areas declined by approximately 0.5–1 log10 levels. After only 8 h of drying, residual virus titers on the bristle fixation were reduced to just above the detection limit of ≤ 102.5 TCID50/ml. In some repetitions, even a value below the detection limit was determined. The experimentally verified viral load on the back of the toothbrush and the bristles also diminished with increasing drying phase, but lasted longer on these areas. After 24 h of drying, the remaining titers on the back of the toothbrush and the bristles were recorded with approximately a 3 log10 loss.It was evident in the experiment that the determined viral load decreases with an increasing drying phase. Moreover, the virus does not retain as long on the bristle fixation compared to the back of the toothbrush and the bristles Fig. . The IAV7.16 TCID50/ml. The remaining virus in the rinsing water led to a mean titer of 106.13 TCID50/ml. After a drying time of 12 h, a titer drop below the detection limit was determined on the toothbrush head.The rinsing of the toothbrush, followed by a drying period of 12 h, showed a reduction of the viral load Fig. . The ave6.23 TCID50/ml. A mean value of 106.23 TCID50/ml was determined in the rinsing water. After a drying time of 12 h, a titer drop below the detection limit was determined on the toothbrush head.As in the case of FeCoV contamination, it was also evident in the case of contamination with IAV H1N1 that rinsing the toothbrush and subsequent air-drying for 12 h resulted in a reduction of the viral load Fig. . The virThe current worldwide pandemic situation shows the danger of the speed of spread of viral pathogens, which should not be underestimated. In the current event of a respiratory virus such as SARS-CoV-2, potential spread cycles must also be recognized on an everyday scale. In this context, dental hygiene plays a role, in addition to the already generally established measures such as personal hand hygiene. Aside of this way of infection, the transmission via contaminated surfaces has been thoroughly discussed . In orde10 (FeCoV) and 2–4 log10 (H1N1) could be detected on all tested toothbrush parts. The residual titer was just above the detection limit at this point and changed little over the remainder of the experiment. Thus, both viruses show a low stability, which is further reduced by rinsing of the contaminated toothbrush parts, where no active virus could be recovered.For this purpose, controlled contamination experiments of different toothbrush areas  were performed to analyze virus tenacity. It was found that the titers of both viruses were rapidly and steadily reduced over the 24 h of the experiment. This reducing effect was particularly rapid on the contaminated toothbrush fixation. Furthermore, already within 12 h, an effective titer reduction of 2.5–5 logMeanwhile, different studies report a certain stability of the infectivity of coronaviruses, especially SARS-CoV-2 on surfaces . InteresCoronaviridae; those viruses have an enveloped spherical structure with a diameter between 60 and 160 nm, while Influenza virus belongs to the family Orthomyxoviridae, having an enveloped pleomorphic structure with a diameter ranging from 100 to 120 nm [FeCoV was selected as an alternative test virus for SARS-CoV-2 in this study. Both viruses belong to the family of In addition to a coronavirus, the current experiment was also performed with another respiratory virus with high clinical relevance: Avian Influenza A virus H1N1. AIV H1N1, which caused a pandemic in 2009 , is a tyIn general, a higher viral load of both viruses on bristles and bristle fixation would have been expected, based on the higher surface size. Nevertheless, the back of toothbrush head was found to show the comparably highest load. This might be explained by its smooth, non-porous surface, allowing a certain stability of the droplets and a slower evaporation, resulting in less collapse of the envelope of the virus. This, however, remains speculative and cannot be finally confirmed by the current data. The toothpaste, brushing technique, the toothbrush type , and interactions with salivatory components may limit the generalizability of the findings.The study only included traditional toothbrush. More in-depth research on dental hygiene routines that deviate from the lab standard applied in this study is needed, for example, with electric toothbrushes or toothbrushes made of different materials  or more frequent brushing cycles.The toothbrush appears to play an insignificant role in the (self-) transmission of coronavirus or influenza virus. Nevertheless, an appropriate use of oral health aids, i.e., using one toothbrush each person, avoid contact between the aids used by different individuals, etc., would be recommendable, regardless of the findings.


Anti-vascular endothelial growth factor is a medicine administered intravitreally by an injection to maintain visual acuity in patients with a variety of retinal diseases. The demand for this treatment has grown considerably in the westernized world the last two decades and will continue to increase due to an aging population. Because of the high volume, injections seize enormous resources and represent high costs for both hospitals and society. Task-shifting of injections from physicians to nurses may be a means to reduce such costs, however the magnitude of possible savings has been poorly investigated. To this end we investigated changes in the hospital costs per injection, six-year cost projections of physician- versus nurse-administered injections for a Norwegian tertiary hospital and we compared the societal costs per patient per year.n = 318) were randomized to either physician- or nurse administered injections, and data were prospectively collected. Hospital costs per injection were calculated as the sum of training costs, personnel time and running expenses. The number of injections for the years 2014 – 21 from a Norwegian tertiary hospital was combined with age group specific injection prevalence and population projections to calculate cost projections for 2022 – 27. Societal costs per patient were calculated as the sum of hospital costs, transport costs for patients, caregivers’ use of time, costs of ophthalmology consultations and community-based homecare.Patients (p = 0.398).The hospital costs per injection were 5.5 € higher for physicians compared to nurses (281.6 € versus 276.1 €). Cost projections estimated an annual hospital saving of task-shifting of 48 921 € for 2022 – 27. Societal costs per patient did not differ significantly between the two groups . Injections with anti-vascular endothelial growth factor (anti-VEGF) have become the most common intraocular procedure worldwide due to its considerable effect on several retinal diseases such as age-related macular degeneration (AMD), retinal vein occlusion (RVO) and diabetic macular oedema (DMO) . The chrphysician = 9.1, SDnurse = 8.1). The patient baseline characteristics are shown in Table 318 patients were recruited as part of a randomized controlled trial examining safety of task-shifting injections from physicians to nurses. Further description of the recruitment and randomization of patients has earlier been published . The stu2◦¹7 = 9.3295 NOK2◦¹7. All costs were adjusted to represent costs in 2017 [Detailed cost calculations were performed, which included the application of a range of unit costs Table . The basWe estimated hospital costs per injection administered by physicians and nurses, respectively. Hospital costs included costs of training, time cost for personnel, clinical support given by a consultant , secretary and other support personnel, equipment including medicine, and running expenses of the premises. While the three first cost categories  differ between physicians and nurses, the last three were fixed costs and equal for both personnel groups.Before the StO-study, all treatment were administered by physicians in training (residents) who spent six months of their residency administering injections. Before nurses were introduced to injection administration, a 10-day training program was designed . Wage cost per day was then divided by the average number of injections per day, giving the personnel costs per injection Table .Table 4HDuring the study period, certified physicians and nurses had the opportunity to seek advice from a consultant. The number of questions asked during the StO-study period was multiplied by the number of minutes used by the consultant and the consultant’s wage cost per minute, giving a total cost. This total cost was divided by the number of injections administered during the StO-study period, and this gave the clinical support costs per injection for physicians and nurses, respectively Table .At the injection clinic, a secretary worked 50%, and in addition, one nurse prepared patients and one nurse assisted injections. Wage costs for support personnel per year were divided by the number of injections administered per year, to find the support personnel costs per injection Table .Information on which anti-VEGF medicine  was administered to each patient was obtained from the EPR, and the unit prices were obtained from the hospital pharmacy. The medicine costs for all patients were summed and divided by the number of injections to find the average medicine cost per injection. The equipment costs included the aggregated volume of disposable equipment at purchase price and the cost of cleaning reusable equipment, divided by the number of injections Table .The cost of the injection clinic premises was based on the area used and included expenses covering cleaning, electricity, and rental costs. Information about injection clinic area was obtained from the hospital's real estate department. Table . The monTo examine future cost consequences of task-shifting, we developed a simple model for projecting the number of injections per year for St. Olavs Hospital for the period 2022–27. For this purpose, the number of injections for 2014–21 was first obtained from the St. Olavs Hospital’s computer system . For this period, we calculated the annual increase of injections per 1000 inhabitants in the St. Olavs population. We assumed this increase to represent an increase in the prevalence of retinal disease (patients per 1000 inhabitants) since injection per patient was presumed to be constant. The mean increase per year was 4.1%. Next, we used the number of injections administered in the StO-study to calculate age-group-specific injection prevalence for the year 2016, which is used as the reference year because this was the mid-year of the StO-study. We used five-year age groups from 30–89 years and 10-year from 90–99.Population numbers and projections from Statistics Norway  were obtTo calculate potential hospital cost savings, the number of injections in 2014–27 was multiplied by the difference in the hospital cost per injection between physicians and nurses, respectively.Societal costs per patient included hospital costs per injection (earlier described), other healthcare costs, transport costs and costs for caregiver’s time. Hospital costs per injection and transport costs per injection were multiplied by the number of injections for each patient to estimate the cost per patient.Other healthcare costs were eye—and homecare costs. Eyecare costs were defined as costs for a consultation either at St. Olavs Hospital or at the patient’s local eye clinic Table . The numHomecare costs was specified as costs for community-based homecare per year. The patient’s homecare use was registered at baseline and the end of the StO-study. The weekly costs were calculated by multiplying mean hours per week with the wage cost per hour of homecare personnel Table . AggregaTransport costs were calculated as the sum of travel costs (to eyecare and injections) for the patient and costs of the caregiver’s time of travelling plus waiting. The travel costs for the patient and the caregiver were calculated by measuring the distance and travel time from home to hospital and from home to eyecare with a web-based map . InformaDescriptive statistics were used to present cost data. To compare the cost per patient between physicians and nurses, Mann–Whitney U tests were used. When calculating eyecare, homecare and transport costs, there were ≤ 0.01% missing values, and calculations were made for the complete cases. For other calculations, there were no missing values. P-values < 0.05 were considered statistically significant. Data preparation and statistical analyses were performed using IBM SPSS software 28.0 , and Microsoft Excel  was used for the cost projections.Written informed consent was obtained from all patients. The study was approved by the Regional Committee of Ethics in Medical Research (2014/1719) and adhered to the Declaration of Helsinki. The study protocol was registered at ClinicalTrials.gov (NCT02359149).per person were lower for a physician compared to a nurse. However, over 10 years, a physician will be expected to administer 2875 injections, and a nurse 9583, resulting in higher physician training costs per injection.In Table The total hospital costs per injection were 5.5 € higher for physicians compared to nurses due to higher wage and training costs per injection Table .The observed number of injections for 2014–21 for St. Olavs Hospital and predicted annual injections for 2022–27 are shown in Fig. An assumed annual increase in injection prevalence of four per cent implies an estimated 10 602 injections in 2027 compared to the observed 6817 in 2021. Applying a more modest growth factor of two per cent implies an estimated 9 374 injections in 2027. By combining annual injection numbers and the difference in hospital cost per injection in favour of nurses, we estimated annual costs saved for St. Olavs Hospital by task-shifting to nurses. The annual mean saving for the period 2017–21 was 33 705 € which in sum for the period was 168 527 €. The estimated total and potential cost savings for the period 2022–27 were 293 527 € (four per cent growth) and 258 017 € (two per cent growth), with an annual mean of 48 921 € and 43 003 €, respectively  were 1920 € (SD 748) per patient. Other healthcare costs per patient were 2169 € (SD 5531), and the transport costs were 1093 € (SD 1073). The societal costs per patient were 5182 € (SD 5777) for the StO-study year, which means that hospital costs per patient accounted for 37% of the societal costs.In this study, the hospital cost per injection was estimated to be 5.5 € lower for nurses than physicians. Applying the estimated cost difference per injection and the projected number of injections, a potential hospital cost saving was found to be between 258 017 € and 293 527 € for the next six years if task-shifting the injections to nurses. We found no statistically significant difference in societal costs per patient between the two groups.The hospital costs per injection were lower for nurses than physicians due to nurses’ lower wages. In St. Olavs Hospital, the wage of a certified nurse is 77% of the wage of a certified physician. The cost savings could be more substantial in countries where the wage gap is more prominent. For instance, in Singapore Teo et al. found a 61% reduction in cost per injection after task-shifting to nurses . The cosThe projected annual mean saving from task-shifting for St. Olavs Hospital from 2022–27 was between 43 003 and 48 921 € (two and four per cent growth). This amounts to half of the wage costs for a full-time nurse position per year. Hence, one can argue that the cost savings of task-shifting the injections to nurses are minor. However, more personnel trained equals more flexibility for the management and is also associated with higher treatment rates . The phySeveral factors influence the future need for injections and thus the calculation of six-year cost projections. Increased awareness of retinal diseases in the population and increased use of advanced diagnostic imaging tools could increase the demand to some extent. Furthermore, novel drugs licensed for other retinal diseases , 24 likeThe difference in societal costs per patient between the physician and nurse group was not statistically significant. Patients randomized to nurses travelled more often with a caregiver. However, the use of a caregiver decreased in the nurse group and increased in the physician group. There are more women in the nurse group, and there are reports that women more often utilize healthcare services and travel more with caregivers , 29. ThiThe mean travel cost per patient per year of all study patients was 1093 €, nurse and physician groups combined, a sum that is equivalent to 1/5 of the total societal cost per patient. The large number of patients receiving injection results in high travel costs for hospitals and society. For instance, in 2016 St. Olavs Hospital treated around 900 patients annually, with the estimated travel costs from the present study amounting to over 1 million euros. A study from Portugal found that the injection rates were negatively associated with travelling distances from home to hospital, meaning that patients that live farther off from the hospital are less likely to receive injections . A NorweThe cost calculations had some limitations. We used a mean medical cost in the calculations since the actual medical costs of the three different medicines, are classified information. A further examination of medicine costs would have been of interest since the cost of various anti-VEGF-agents differ considerably and therefore choice of anti-VEGF-agent will likely affect hospital costs as such. However, when we compared injections by nurses versus physicians, we assumed equal distribution of anti-VEGF agents in both groups in accordance with randomization. It follows that a different choice will not affect the difference between nurses and physicians, only the level of hospital costs. We calculated time costs and clinical support costs for injections for the two groups, but available data did not enable the calculation of such costs for individual patients. Hence variation within groups remained unknown. A third limitation was the uncertainty when projecting the future number of injections. Pharmaceuticals are developing new long-acting anti-VEGF medicines , and altThe strength of our study was the comprehensive calculation of hospital costs, covering most relevant cost categories. Moreover, the detailed calculation of injection costs like training, time costs and clinical support where data collected in the StO-study.To the best of our knowledge, the present study is the first to compare costs of physician- versus nurse-administered injections in a Nordic setting. Task-shifting the treatment to nurses provided a modest cost saving per injection for the hospital in addition to increased flexibility when reallocating physician resources from injections to other tasks. The societal cost per patient was not significantly different for physician- and nurse-administered injections. However, transport costs are high because of the long travel distances for patients in central Norway. Reducing transport costs by minimizing the number of clinical visits and organizing consultations and injections on the same day, may be a solution to reduce costs for society in the future.


Discovered as a contaminant of adenovirus stocks in the 1960s, adeno-associated virus (AAV) is a mono-stranded DNA virus that depends on helper factors to replicate. Even though AAV is endemic in the human population (35–80%), it is remarkable that many issues concerning the natural infection by this virus remain unanswered. In this study, we reflect on the main basic aspects of AAV biology and provide an overview of the studies exploring the impact of AAV infection on human health, focusing on three major research areas including, (i) cervical and (ii) liver cancer, and (iii) reproductive system disorders. Conflicting results have been obtained into the association of AAV infection with the occurrence of adverse reproductive outcomes, such as placental complications, spontaneous abortion, and fertility disorders, or with a protective role in HPV-related cervical carcinogenesis. Noteworthy, recent reports have identified AAV insertional mutagenesis as a novel risk factor for the development of hepatocellular carcinoma. This latest finding raises concern regarding the widespread usage of AAV vectors in liver-targeted gene therapy. Adeno-associated virus (AAV) was discovered as a contaminant in a simian adenovirus type 15 preparation in 1965 . Since tParvoviridae family, AAV belongs to the Dependoparvovirus genus. There are at least 13 naturally occurring serotypes, each with a different tissue tropism . Nevertheless, considering the extensive usage of rAAV vectors in liver-targeted gene therapy and its potential for insertion into the human genome, patients treated with rAAV vectors should be followed longitudinally to monitor long-term consequences and determine the risk of HCC development.


Apodemus agrarius.Hemorrhagic fever with renal syndrome (HFRS) caused by hantaviruses is a frequently reported acute hemorrhagic fever in South Korea. These viruses are transmitted by various rodent species such as A. agrarius, 49 Crocidura lasiura, and 24 Myodes regulus. Nested RT-PCR was performed to examine the rate of hantavirus infection in wild rodents, and 1.88% (11/585) of all rodents, 1.17% (6/512) of A. agrarius, 6.12% (3/49) of C. lasiura, and 8.33% (2/24) of M. regulus tested positive. The nucleotide sequence analysis of the eleven PCR-positive products revealed that six PCR products showed over 85% sequence similarity with the Jeju virus, four showed over 99.7% similarity with the Hantaan virus, and one showed over 95.3% homology with the Imjin virus. Moreover, IgG antibodies against the Hantaan virus were detected in 6.15% (36/585) of all rodents, 6.8% (35/512) of A. agrarius, and 4.17% (1/24) of M. regulus. IgG antibodies were not detected in C. lasiura.To investigate hantavirus infection and seroprevalence in rodents, wild rodents were captured from two districts in the suburbs of Gwangju Metropolitan City from January 2016 to December 2018. Nested reverse-transcription polymerase chain reaction (RT-PCR) targeting the hantavirus-specific L segment and indirect immunofluorescence antibody (IFA) assay using Hantaan virus antigen slides were performed. A total of 585 wild rodents were captured—512 A. agrarius, C. lasiura, and M. regulus captured in the suburbs of Gwangju Metropolitan City, South Korea, and it was demonstrated that they were various strains of hantaviruses such as the Hantaan, Jeju, and Imjin viruses.Hantaviruses were detected in all three wild rodent species of  Hantaviridae, genus Hantavirus) are rodent viruses that can cause two fatal human diseases—hemorrhagic fever with renal syndrome and hantavirus pulmonary syndrome. These viruses are transmitted by various rodent species such as Apodemus agrarius. In the present study, wild rodents captured in the suburbs of Gwangju Metropolitan City were analyzed by nested reverse-transcription polymerase chain reaction targeting the hantavirus-specific L segment and indirect immunofluorescence antibody assay using Hantaan virus antigen slides. A total of 585 wild rodents were captured and 1.88% (11/585) of all rodents, 1.17% (6/512) of A. agrarius, 6.12% (3/49) of Crocidura lasiura, and 8.33% (2/24) of Myodes regulus were PCR-positive. Of the eleven PCR-positive wild rodents, six PCR products showed over 85% sequence similarity with the Jeju virus, four showed over 99.7% similarity with the Hantaan virus, and one showed over 95.3% homology with the Imjin virus. Moreover, IgG antibodies against the Hantaan virus were detected in 6.15% (36/585) of all rodents, 6.8% (35/512) of A. agrarius, and 4.17% (1/24) of M. regulus. IgG antibodies were not detected in C. lasiura. When we examined the detection rate of hantavirus genes in different seasons, hantaviruses were most commonly detected in fall (seven cases [5.69%]) and winter (four cases [2.76%]). No hantaviruses were detected in spring or summer. However, the seasonal prevalence of IgG antibodies was higher in spring and summer  than in fall and winter . This study was performed on a monthly basis throughout a three-year period, and thus, it provides reliable data that may provide insight into preventive measures against HFRS. Our results suggest that various types of hantaviruses, including Hantaan, Jeju, and Imjin viruses, are distributed throughout Gwangju Metropolitan City, South Korea. Data regarding the rate of Hantaan virus infection in rodents in Gwangju and knowledge of the hantavirus seroprevalence, species, and genotypes circulating in these domestic rodent species will provide useful information for developing vaccines and diagnostic testing using sequence data. These will help increase preparedness for the emergence of new species.Hantaviruses (family  Hantaviridae, genus Hantavirus) are rodent viruses that can cause two fatal human diseases—hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS) ) and winter (four cases [2.76%]). No hantaviruses were detected in spring or summer.In this study, we performed nested RT-PCR targeting the hantavirus L segment and found three hantaviruses, namely, the Jeju, Hantaan, and Imjin viruses, in wild rodents. In nested RT-PCR performed to examine hantavirus genotypes, 11 of 585 samples (1.88%) were PCR-positive, and hantavirus genes were detected in all the three species—When the IgG cut-off value (≥1:16) was used in the IFA, 36 of 585 serum samples (6.15%) tested positive for IgG antibodies against hantaviruses. Two of 36 IFA IgG-positive rodents were PCR-positive for hantaviruses. In particular, the seasonal prevalence of IgG antibodies was higher in spring and summer  than in fall and winter . The antibody positivity rate identified in this study was slightly lower than or similar to the rates reported by Song et al. and Nam et al.  ) and M. regulus (17.2% [5/29]) ) than in spring (23.1% [87/376]). When examining the IgG antibodies by rodent species, (21.4% 3/) and M. In 2011, Ryou et al. performed PCR targeting the S segment of the Hantaan virus in wild rodents captured from five provinces in South Korea and reported a PCR-positivity rate of 3.3% and varying positivity rates of Hantaan virus antibodies from 4–29% in different regions. In addition, they reported high antibody prevalence between March, May, October, and December and that the seasonal pattern of Hantaan virus positivity in rodents affected the seasonal prevalence of HFRS in humans .C. lasiura, two in A. agrarius, and one in M. regulus. Of the hantaviruses that showed homology with Hantaan virus, three were detected in A. agrarius and one in M. regulus. One hantavirus that showed homology with the Imjin virus was detected in A. agrarius.Of the 11 hantaviruses detected in rodents in this study, six showed over 85% sequence similarity with the Jeju virus, four showed over 99.7% sequence similarity with Hantaan virus, and one showed 95.3% sequence similarity with the Imjin virus. Of the hantaviruses that were homologous to the Jeju virus, three were detected in C. lasiura (Ussuri white-toothed shrew) that had been captured in the demilitarized zone of South Korea. Seven cases of Imjin virus were confirmed by RT-PCR out of a total of 115 C. lasiura individuals captured. The prevalence of Imjin virus was highest in the fall season based on the IFA assay and RT-PCR test results [C. lasiura individuals captured in the Republic of Korea between 2004–2010 [C. lasiura, so the major animal host of Imjin virus was reported as C. lasiura.In 2009, Song et al. reported Imjin virus, a novel hantavirus, in the lungs of  results . In addi004–2010 . No viruCrocidura shantungensis) that were captured on Jeju Island, South Korea. Eight cases of Jeju virus were confirmed by RT-PCR among the 51 C. shantungensis captured on Jeju Island. But the virus was not found in the 28 C. shantungensis individuals that had been captured on mainland Korea. Moreover, there is a divergent ancestral lineage between the Imjin and Jeju viruses despite the close phylogenetic relationship between the two reservoir hosts [In 2012, Arai et al. detected a novel hantavirus, Jeju virus, in Asian lesser white-toothed shrews (ir hosts .Apodemus agrarius), Seoul virus in brown rat (Rattus norvegicus), Soochong virus in Korean field mice (Apodemus peninsulae), and Muju virus in royal voles (Myodes regulus) [A. agrarius, C. lasiura, and M. regulus may transmit hantaviruses to humans.Hantaviruses detected in rodents captured in South Korea include Hantaan virus in striped field mice (regulus) . The resThe high PCR positivity rates of hantavirus in wild rodents captured in fall and winter somewhat closely match the seasonal detection rates reported by Park, who analyzed 6,132 HFRS cases recorded by the Korea Centers for Disease Control from 2002 to 2016 at a national scale . He repoC. lasiura, A. agrarius, and M. regulus—other than the previously reported C. shantungensis host suggests high infection prevalence and frequency of spillover events. This study is the first to present such findings. The same goes for the detection of Imjin virus in A. agrarius. Unlike previous studies that were conducted during a specific period of the year, this study was performed on a monthly basis throughout a three-year period, and thus, it provides reliable data that may provide insight into preventive measures against HFRS. The rate of Hantaan virus infection in rodents in Gwangju and the information about various hantavirus strains presented in this study will be useful for the development of vaccine candidates and preventive measures for HFRS.The results of this study are meaningful in that at least three hantavirus strains, namely Hantaan, Jeju, and Imjin viruses, were detected using nested RT-PCR targeting the hantavirus L segment and that the prevalence rate for Hantaan virus-specific IgG antibodies was measured in rodents captured within a limited area, namely, Gwangju Metropolitan City, South Korea. Above all, the detection of Jeju virus in three species—In conclusion, potentially pathogenic hantaviruses were detected in all three rodent species that transmit HFRS in Gwangju Metropolitan City. Various types of hantaviruses, including Hantaan, Jeju, and Imjin viruses, are distributed throughout Gwangju. Knowledge of the hantavirus seroprevalence/ species/genotypes circulating in these domestic rodent species will provide an useful information for the rapid diagnosis of human infections as well as the potential for emergence of new species.


Drawing from a review of the literature and international statistical sources, this paper identifies the global prevalence of young onset dementia (YOD), its complex aetiology and its social and economic costs. YOD is often defined as dementia affecting those aged 65 or under, although definitions vary. Prevalence estimates also vary but recent evidence suggests a rate in the region of 119.0 per 100,000 population for those aged 30-64, and this is expected to increase markedly in the coming decades. Research identifies that the causes of YOD vary greatly among younger people. Individuals experiencing YOD face a complex range of psychological, familial, and socioeconomic challenges, while often being active in the labour market at the time of diagnosis. Costs of YOD cannot be easily disentangled from those for dementia generally but include: the direct costs of the ‘resources’ drawn on by the person with dementia; indirect costs, usually considered in terms of the value of lost productivity by the person with dementia or their carer, and less tangible costs, such as in terms of psychological wellbeing, social exclusion and time costs. The paper concludes by considering the potential significance of YOD for the future labour market in the context of ongoing efforts to prolong working lives.


Surveys were conducted in 2016 and 2017 across the main cassava‐growing regions of Burkina Faso to assess the status of cassava mosaic disease (CMD) and to determine the virus strains causing the disease, using field observation and phylogenetic analysis. CMD incidence varied between regions and across years but was lowest in Hauts‐Bassins  and highest in Centre‐Sud  and in Boucle du Mouhoun . The lowest CMD severity was found in Est region (2.0) for both years and the highest in Sud‐Ouest region  and Centre‐Sud region . The CMD infection was primarily associated with contaminated cuttings in all regions except in Hauts‐Bassins, where whitefly‐borne infection was higher than cuttings‐borne infection in 2016. PCR screening of 687 samples coupled with sequence analysis revealed the presence of African cassava mosaic‐like (ACMV‐like) viruses and East African cassava mosaic‐like (EACMV‐like) viruses as single infections at 79.5% and 1.1%, respectively. Co‐infections of ACMV‐like and EACMV‐like viruses were detected in 19.4% of the tested samples. In addition, 86.7% of the samples positive for EACMV‐like virus were found to be positive for East African cassava mosaic Cameroon virus (EACMCMV). Phylogenetic analysis revealed the segregation of cassava mosaic geminiviruses (CMGs) from Burkina Faso into three clades specific to ACMV, African cassava mosaic Burkina Faso virus (ACMBFV), and EACMCMV, confirming the presence of these viruses. The results of this study show that EACMCMV occurrence may be more prevalent in Burkina Faso than previously thought. Cassava mosaic disease is widespread in Africa. In Burkina Faso, we assessed the main epidemiological parameters and characterized the major viruses associated with the disease. CMD is a major constraint to cassava production, which causes tuber yield losses estimated at $2.7 billion annually  coordinates were recorded for each field.2.2Taq DNA polymerase (GeneON), and 150 ng DNA template of each sample. The DNA amplification was carried out in a SimpliAmp thermal cycler (Life Technologies Holdings Pte Ltd). The PCR temperature profile was set at 94°C for 4 or 5 min for initial denaturation, followed by 35 cycles of amplification at 94°C for 45 or 60 s, 55°C for 45 or 60 s, and 72°C for 55 or 60 s (depending on primers). The final elongation step was performed at 72°C for 7 or 10 min. PCR‐amplified products were subjected to 1% agarose gel electrophoresis, stained with ethidium bromide. The electrophoresis was performed at 100 V and the gel was visualized using a Compact Digimage System, UVDI series (MS major science). PCR products of 40 ACMV‐like positive samples (randomly selected from the regions) were directly sequenced in both forward and reverse orientations using the Sanger method at Inqaba Biotec company (South Africa) to determine their identity. PCR products of 15 EACMCMV positive samples were also subjected to sequencing in both forward and reverse orientations to confirm their identity.Total DNA was extracted from cassava leaves using the CTAB protocol as previously described .Data analysis was performed using the R software v. 3.6.1 (R Development Core Team). The normality of the variables was determined using the Shapiro–Wilk test. When the variable was not distributed according to the normal distribution, the generalized linear model was used. The difference in the number of whiteflies per plant between regions and the difference in the severity score of CMD between regions in the same year were assessed using the generalized linear model and Tukey's pairwise mean comparison test. The difference in the number of whiteflies per plant and the difference in the severity score of CMD between 2016 and 2017 were assessed using Wilcoxon test with continuity correction. A test of pairwise comparison of proportions was used based on a G‐test with correction of BY .The amplicon sequences were trimmed and assembled de novo using Geneious v. 8.1.7 (Biomatters Ltd) software. Consensus sequence obtained from forward and reverse sequences for each sample was subjected to BLASTn in NCBI for preliminary species assignment and subsequently for pairwise sequence comparison . Indeed, compared to 2016, no cassava fields were found to have CMD symptoms in the provinces of Nahouri (Centre‐Sud region) and Bougouriba (Sud‐Ouest region) in 2017. Typical CMD symptoms observed across farmers’ fields included distinctive leaf mosaic symptoms often associated with leaf distortion and reduction  of surveyed localities, with the lowest proportion  of infected fields in the region of Hauts‐Bassins. Cassava fields in 65.9% (29/44) of localities showed CMD symptoms in 2017, with the lowest proportion  in Centre‐Est region . In 2016, the overall CMD incidence across the surveyed fields in Burkina Faso was 11.3% (216/1920) and ranged from 6.0% (36/600) in Hauts‐Bassins region to 18.5% (50/270) in Centre‐Sud region. In 2016, the difference between the lowest incidence and the highest incidence was highly significant (p ≤ 0.001). For the 2017 survey, the overall CMD incidence was 18.9% (329/1720). The lowest incidence in 2017 was observed in the Hauts‐Bassins region with 5.4% (21/390), whereas the Boucle du Mouhoun region was observed to have the highest incidence 51.7%  were found between CMD symptom severity scores in 2016 and 2017 in three regions . In most of the regions, no significant difference was found between the proportion of plants with different CMD symptom severity scores in 2016. In 2017, the proportion of plants with CMD symptom severity score 2.0 was higher than the other symptom severity scores in most of the regions, whereas in the region of Centre‐Sud the proportion of plants with CMD symptom severity score 3 was the highest  to 3.3 (Sud‐Ouest region) in 2016. In 2017, the mean CMD symptom severity score was 2.5 with the lowest severity in Est region (2.0) and the highest severity in the Centre‐Sud region (2.8) Figure . Signifit Figure .3.3Determination of whitefly counts and distribution was conducted at the time of the CMD incidence and severity survey to ensure the parameters that might affect the epidemiology of CMD in the field were the same, and the plants were of similar age. The methods have been harmonized across 10 West and Central African countries, Sierra Leone, Ivory Coast, Burkina Faso, Ghana, Nigeria, Benin, Togo, Cameroon, Gabon, and Democratic Republic of Congo, to ensure the data is comparable. The adult whitefly counts were very low in 2016 and a similar trend was observed in 2017, with a mean of 0.1 and 0.7 per plant in 2016 and 2017, respectively. The highest mean whitefly count was observed in the region of Boucle du Mouhoun (1.08) in 2016. In 2017, the highest mean whitefly count (2.7) was observed in the regions of Boucle du Mouhoun and Cascades. In most of the regions, the mean number of whiteflies per plant was higher in 2017 than in 2016 Figure .When the number of plants with symptoms infected through cuttings or by whitefly transmission were compared, a preponderance of cutting‐borne infections was detected in 2016  and 2017 . The exception was Hauts‐Bassins region, where greater whitefly‐borne infections were recorded in 2016 Figure . It is l3.4A total of 687 cassava leaf samples were collected from 452 plants with symptoms and 235 plants without symptoms in 2016 and 2017 for PCR analysis. Among the samples having observable symptoms, 4.0% (18/452) tested negative for ACMV‐like virus and EACMV‐like virus. On the other hand, 2.1% (5/235) of symptomless samples tested positive for ACMV‐like virus. Approximately 63.9% (439/687) of collected samples tested positive for CMGs. Among the positive samples, the single ACMV‐like virus infection was by far the most frequent, accounting for 79.5% (349/439) of all infection, followed by mixed infections of ACMV‐like virus and EACMV‐like virus with 19.4% (85/439), and single infection of EACMV‐like virus with 1.1% (5/439). The single infection of ACMV‐like virus was predominant in all surveyed regions, with the highest proportion (100%) in the Centre‐Est and Est regions. The mixed infection occurred in the remaining six regions, with the highest proportions in Centre‐Sud  and Sud‐Ouest  regions. The single infection of EACMV‐like virus was found in the regions of Sud‐Ouest , Cascades  and Centre‐Sud  but no significant difference was found between these proportions  showed that the sequences of the 40 samples that tested positive for the ACMV‐like virus were most closely related to ACMV and ACMBFV. Indeed, they shared the highest nucleotide identity (98%–99%) with ACMV isolates from Ghana , Ivory Coast (AF259894), Burkina Faso (FM877473), and Nigeria (MH251339), and with ACMBFV isolates from Burkina Faso . The sequences of the 15 samples that tested positive for EACMCMV were most closely related to the EACMCMV and shared the highest nucleotide identities (97%–98%) with isolates from Ghana (MG250164), Ivory Coast (AF259896), Nigeria , and Madagascar (KJ887944). The ML phylogenetic tree inferred from alignment of coat protein (CP) gene sequences from Burkina Faso (25 ACMV‐like virus and six EACMCMV) and other CMGs confirmed that the sequences from Burkina Faso are phylogenetically associated with ACMV, ACMBFV, or EACMCMV , which shows that the viruses can be latent in the plants without manifesting symptoms. Therefore, the use of symptomless cassava landraces as an option to manage CMD could inadvertently result in increased cutting‐borne transmission because they may harbour CMGs. We propose that the use of certified virus‐free cuttings for the establishment of new cassava fields will be crucial for fighting the transmission of CMD. In the absence of certified virus‐free cuttings, the training of farmers on how to select healthy cuttings for the new planting season and on use of in‐field diagnostic applications will be crucial to bring down the incidence or transmission of these viruses of high economic importance.The authors declare that they have no conflict of interest.


The immobilization process was confirmed by atomic force microscopy (AFM). The prepared aptamer/MXene immobilized on AuMGE can detect the Zika virus through capacitance change according to the target concentration. The capacitance signal from the biosensor increased linearly according to increment of envelope proteins in the human serum. The limit of detection was determined to 38.14 pM, and target proteins could be detected from 100 pM to 10 μM. Thus, the developed electrical aptabiosensor can be a useful tool for Zika virus detection.Zika virus is a highly infectious virus that is part of the flavivirus group. Precise diagnosis of the Zika virus is significant issue for controlling a global pandemic after the COVID-19 era. For the first time, we describe a zika virus aptamer-based electrical biosensor for detecting Zika virus in human serum. The electrical biosensor composed of a Zika virus aptamer/MXene nanoparticle heterolayer on Au micro-gap electrode (AuMGE)/print circuit board (PCB) system. The Zika virus aptamer was designed to bind the envelope protein of the Zika virus by systematic evolution of ligands by exponential enrichment (SELEX) technique. The binding affinity of the aptamer was determined by fluorescence. For improving the sensor signal sensitivity, TiThe online version contains supplementary material available at 10.1186/s40580-022-00332-8. Flavivirus which also include viruses such as yellow fever, dengue fever, and West Nile. Zika virus was discovered in 1947 and was first recognized as a human disease when a viral infection was confirmed in humans in Nigeria in 1953 [Zika is a mosquito-borne virus of the genus  in 1953 . In the  in 1953 . TherefoMethods of diagnosing Zika virus infection include detection of viral nucleic acid by reverse-transcription polymerase chain reaction (RT-PCR) , detectiSeveral studies have focused on aptamers in biosensor fabrication as a useful bioprobe , 12. WheThe Zika virus aptamer was synthesized using the SELEX technique and applied to a capacitance-based biosensor composed of DNA aptamer/MXene on Au micro-gap electrode (AuMGE) to detect Zika virus in human serum samples. The chemically produced aptamer is a single-stranded nucleic acid that can specifically bind to the Zika virus envelope proteins. MXene was used to increase the detection sensitivity. MXene is a plate-shaped nanomaterial composed of transition metal carbides and nitrides, and exhibits easy processability, high charge storage capacity, high mechanical strength, and excellent electron/ion conductivity . In addiSince the AuMGE provides a small active area, it is possible to measure with a small amount 10 μl) of sample, and the test can be repeated up to 28 times with one electrode. Furthermore, MXene-based biosensors are utilized as effective transducers for immobilizing biological receptors on surfaces and have proven to be highly sensitive and selective sensing platforms –20. Figu0 μl of sFor performing SELEX, the screening of Zika envelope aptamer was carried out by procurement of XELEX DNA Core kit from EURx (Poland) based on previous research . The DNAThe development of Zika envelope-specific aptamers is performed based on the SELEX procedure. In this study, the existing SELEX procedure, which is performed based on the binding of a His tag-existing target protein with a cobalt bead, was modified. In the sample preparation step, the reaction between the target protein and Sulfo-NHS-LC-biotin binds biotin to the protein surface based on the S–S bond. Proteins modified with biotin The sub-ssDNA library and DNA stabilization buffer used in the experiment were purchased and used by the XELEX DNA Core kit .In this study, the SELEX process consists of four steps: protein sample preparation, binding of ssDNA to the target protein, capture and isolation of ssDNA, and amplification through PCR. The concentration of target protein, reaction temperature and time, and PCR conditions were set based on the optimization performed in previous studies . Additio2, which are components of the kit. The DNA library in the kit consists of the sequence 5′-TGA CAC CGT ACC TGC TCT-N40-AAG CAC GCC AGG GAC TAT-3′; the forward primer is 5′-TGA CAC CGT ACC TGC TCT-3′, and the reverse primer is 5′-ATA GTC CCT GGC GTG CTT-3′. The protein-bound streptavidin magnet beads and 200 μl of DNA library diluted with SELEX buffer were reacted for 1 h, and the reaction product was washed 5 times using the buffer. Then the beads bound with DNA were collected using a magnet. The Zika envelope protein was denatured by heat-treating the sample in an 80 °C thermostat for 10 min. Finally, beads and denatured proteins were removed by centrifugation, and template DNA from the supernatant was collected.In the sample pretreatment step, 50 μl of Zika envelope protein diluted to a concentration of 0.25 mg/ml and 10 μl of 2 mM Sulfo-NHS-LC-Biotin  were stirred at room temperature at high speed for 1 h to biotinylate the Zika envelope surface. Unreacted biotin was removed by centrifugation for 20 min at 4000 rpm using a 3000 NMWL Amicon centrifuge filter . Next, the protein on the top of the filter was recovered, 100 μl of Streptavidin mag net beads  were added, and the mixture was stirred for 1 h. Subsequently, the unreacted Streptavidin magnet beads were washed out 3 times using SELEX buffer containing NaCl, KCl, Tris, and MgClPCR master mix containing DNA polymerase, Zika aptamer, and thiol-modified Zika aptamer were purchased from Bioneer (South Korea). Amplification of template DNA through PCR was performed as follows: denaturation at 94 °C for 3 min; annealing at 55 °C for 30 s; and amplification at 72 °C for 1 min. The PCR cycle was repeated 10 times.From the second SELEX round, it is performed using the ssDNA pool amplified by the PCR process in the previous round instead of the sub-ssDNA library. To confirm the reactivity of the amplified ssDNA pool and the target Zika envelope protein, 8% TBE-PAGE was performed in rounds 4 and 10 using the ssDNA pool. The binding of the ssDNA pool to the zika envelope protein in round 10 significantly changed the DNA band compared to the control . In addition, the dG values for the secondary structures are shown in Table Secondary structures and dGs of the four candidate sequences selected in session 2.2 were predicted prior to measuring binding affinity. The secondary structure of the predicted sequence is shown in Additional file + ions and 5 mM Mg2+ ions and the room temperature conditions of 25 °C through the mFold application (www.unafold.org), is shown in Additional file Before measuring binding affinity, the secondary structure and dG of the four candidate sequences selected in session 2.2 were predicted. the secondary structure of the predicted sequence, assuming the salt conditions of 140 mM NaBinding affinity measurements are performed with a bead-based fluorescence binding assay. Fluorescein phosphoramidite (FAM), a fluorescent substance, was added to the 5' ends of the Zika-7 sequence and the Zika-25 sequence, and diluted to 3 nM, 30 nM, 300 nM, 1 μM, and 1.5 μM. Then, each concentration of aptamer was incubated with a magnet bead to which Zika envelope protein was fixed after the sample pretreatment step of Session 2.2 for 1 h. Afterwards, the magnet bead was washed 5 times with a washing buffer to remove unbound aptamers, denatured proteins by heat treatment at 80 °C, and centrifuged to recover aptamers. The collected fluorescent aptamers were measured for absorbance at 480 nm and emission at 528 nm using a Bio Tek Synergy LX multimode reader (USA). The fluorescence intensity was plotted according to the aptamer concentration, and Kd was calculated through a calibration curve based on the adsorption isotherm equation.2 layer, and silane was activated by heating in an oven at 70 °C for 1 h. Thiol-modified (SH–) groups were generated by reacting 10 μl of 6-mercaptohexanoic acid  and activated AuMGE for 1 h. Then, 10 μl of 1 mg/ml MXene was added and maintained at room temperature for 12 h for the reaction to occur. Finally, 10 μl of 5% mercaptopropyltriethoxysilane  was applied for 11 min to create a SH group-modified layer.MXene, synthesized according to a previous protocol , was dil2 gas.SH group-modified Zika virus envelope protein aptamer was diluted to 1 μM with DIW, and 10 μl was reacted with the SH layer of AuMGE for 3 h to deposit the aptamer and bind it through self-assembly. All reactions were carried out at room temperature, and the residue was removed with DIW and N2 bare electrodes, SiO2/MXene electrodes, and SiO2/MXene/Aptamer/Zika envelope immobilized electrodes in non-contact mode using atomic force microscopy . The cantilever used for the investigation was PPP-NCHR  and a silicon tip with a resonant frequency of 330 kHz and a spring constant of 42 N/m was selected. AFM measurement conditions were established based on a previous study [Characterization of biomolecules immobilized on Au electrode surfaces was investigated for SiOus study .Capacitance was measured using an LCR meter  controlled by LabView program. The probe of the LCR meter was connected to a socket on the PCB  where AuMGE was mounted. Capacitance was measured at 10 mV for 10 s at 1 MHz with reference to the previous study .+ ions and 5 mM Mg2+ ions. Based on the predicted dG values, two aptamers were selected. and binding was determined using 8% native-TBE PAGE binding test. Figure After sequencing, 50 aptamers were obtained by SELEX. The secondary structure of the Zika envelope aptamer was predicted by mFold at 25 °C with a buffer composition of 140 mM Nad of the aptamer was calculated based on the fluorescence intensity and the protein concentration according to the aptamer concentration using the following isothermal adsorption Eq. , root-mean-squared (RMS) roughness (Rq), and vertical distance (Vd) of the each sample Fig. a–e. The The AuMGE fabricated in this study consists of a large-area counter electrode and 28 electrodes with a gap of 5 μm. Each electrode is bound to a socket connected to the PCB, and for this purpose, the length of the wire connecting the electrode and the pad has a certain difference. As shown in Additional file ε is the dielectric constant between the electrodes, ε0 is the permittivity of free space, A is the area of the electrodes, and d is the distance between the electrodes. Figure The manufactured bare electrode is immobilized on the electrode surface in the order of MXene, Aptamer, and Protein, and the measured capacitance changes depending on the material to be fixed. The change in capacitance is defined using Eq.  consistiSensor performance evaluation was performed by diluting the Zika envelope protein concentration from 100 pM to 10 μM in six steps. As shown in Fig. The LOD was calculated using the Zika envelope protein sample diluted with 10% human serum. In the same manner as the experimental conditions performed previously, measurements were carried out in six steps, 10 times each, in the concentration range from 100 pM to 10 μM, and the measured results are shown in Fig. This study presented the first Zika virus aptamer that successfully binds the envelope protein of the Zika virus. Moreover, the prepared aptamer was introduced to the electrical biosensor as the bioprobe for Zika virus detection with MXene and AuMGE. In this study, the Zika virus aptamers with high reactivity with Zika envelope protein were selected through a screening process known as SELEX. MXene is a nanoparticle with a high electrical conductivity that is fixed to the electrode gap to expand the reaction area of the electrode and improve sensitivity. The proposed biosensor, based on a combination of the aptamer, MXene, and AuMGE can detect a Zika virus envelope protein at a concentration of 38.14 pM within 10 s in 10% human serum conditions. Moreover, the blind test results showed the prepared aptasensor can detect Zika virus well in undefined samples. Compared with previous studies Table , the autAdditional file 1: Fig. S1 Schematic diagram of SELEX Process. Fig. S2 8% TBE PAGE result of 10 round ssDNA pool. Fig. S3 Structures of candidate aptamers prepared in Table. 1 a Zika -07 aptamer; b Zika-09 aptamer; c Zika-17 aptamer; d Zika-25 aptamer. Fig. S4a 8% TBE PAGE result of Zika 25 Aptamer; b 2D structure of Zika 25 aptamer base; c Binding affinity of Zika aptamer.


Orthopoxvirus genus of the Poxviridae family with a similar clinical appearance to smallpox. The symptoms of monkeypox include fever, headache, muscle aches, and lymphadenopathy. The transmission of monkeypox occurs from infected animals to humans or through direct contact , respiratory droplets, and clothing such as towels. The incidence of monkeypox is rising drastically over the world. This short communication discusses the causes of the rising monkeypox cases and emphasizes strategies to prevent the spread of the virus.Monkeypox is one of the many zoonotic viruses that belong to the  Orthopoxvirus genus of the Poxviridae family, which causes a disease similar to smallpox in humans. Human monkeypox is a brick-shaped, double-stranded DNA virus that attaches to glycosaminoglycans to enter host cells. As an enveloped virus, it may enter host cells by mimicking apoptosis , fever (82 percent), sweats (82 percent), chills (82 percent), a persistent cough 73 percent), lymphadenopathy 55 percent), and sore throat (55 percent) 5 percent. Accordi3 percentth November 2022 suggest that there are over 80,000 cases in 103 non-endemic countries, with approximately 29,080 confirmed cases in the United States alone and a total of 53 deaths globally  as of 17Before 1980, when the World Health Assembly declared smallpox extinct, most nations halted routine smallpox immunization . BecauseOrthopoxviruses infection (The smallpox vaccine is protective against both smallpox and monkeypox. Cross-protective immunity against West African monkeypox can be maintained for decades in the population who have received smallpox vaccination in the distant past . Anothernfection , but vacnfection .Amid the COVID-19 pandemic, countries with a middle income cannot endure another epidemic due to their inadequate healthcare infrastructures and the massive pressure placed on their already overburdened healthcare systems. Insufficient numbers of healthcare experts and poor hospital facilities are inadequate to address the populace's needs. Moreover, as the monkeypox virus is rapidly spreading across countries and creating a synchronized detrimental impact, healthcare officials in countries where the virus is endemic must suggest large-scale smallpox vaccination efforts. Multiple observational studies have demonstrated that cross-immunity with smallpox vaccination is around 85 percent effective in preventing monkeypox . As withThe original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.HS: conceptualization, writing–original draft, final approval, and agreeing to the accuracy of the work. MM and NS: writing–original draft, final approval, and agreeing to the accuracy of the work. All authors contributed to the article and approved the submitted version.


Avian influenza A(H5N8) virus has caused major epizootics in Europe since 2016. We conducted virologic analysis of aerosol and dust collected on poultry farms in France during 2020–2021. Our results suggest dust contributes to viral dispersal, even early in an outbreak, and could be a valuable surveillance tool. Orthomyxoviridae family. Wild aquatic birds are the virus reservoir and generate occasional worldwide panzootic outbreaks during seasonal migrations  in France highly affected by HPAI H5N8 virus outbreaks. On the basis of daily official outbreak reports, we identified HPAI-infected poultry houses and poultry houses in close vicinity or with epidemiologic links to infected houses. The study included a total of 48 duck houses, 12 chicken houses, 2 quail houses, and 1 goose house. We selected farms identified as being near an HPAI outbreak to reflect a range of sanitary statuses and infection stages . We specifically included houses without clinical signs among animals to evaluate virus dispersal and dust testing for HPAI surveillance in the early stages of infection.https://www.bertin-instruments.com) and the NIOSH BC 251 developed by the National Institute for Occupational Safety and Health   . Furtherv/niosh)  Table 1.>1 µm  in houses with or without poultry showing clinical signs, we used a latent class modeling approach, necessary when no standard has been established . Model oFinally, to assess the infectiousness of environmental samples, we processed 25 surface dust or aerosol samples taken from 5 animal houses and used these for virus isolation in embryonated eggs . Among 2We used field conditions to evaluate whether dust from poultry farms contained HPAI viruses and to compare surface dust and aerosol testing for HPAI virus against official swab-based methods. We used wipe tests to collect surface dust and 2 bioaerosol devices to collect aerosol samples during the 2020–21 HPAI H5N8 virus epizootic outbreak in France. Standard molecular analysis detected high viral RNA loads in the early phase of flock infection, before clinical signs appeared. In addition, size fractioning of aerosol samples revealed that high RNA viral loads and infectious viral particles were associated with the largest particles (>1 µm), which are easy to collect and use for molecular analysis. However, the field conditions we used cannot be reproduced in experimental animal trials because of ethical and biosecurity requirements, which result in dramatically lower dust loads than those found in the field.Recent research on influenza transmission routes revealed that nonrespiratory airborne particles are more likely to cause infection than are droplets or fomites  clade 2.3.4.4b virus in dust samples from poultry farms, France, 2021.