Financial protection mechanisms are in place to overcome the costs of a few diseases in India. Our objective was to estimate the costs incurred due to Oral Potentially Malignant Disorders (OPMD) and to determine predictors of such costs.We found that the median (Interquartile range IQR) total costs of OPMD was Indian Rupees (INR) 500 (350–750), direct medical costs was INR 0 (0–50), direct non-medical costs was INR 150 (40–200) and indirect costs was INR 350 (250–500). The travel cost to attend the health facilities [INR 100 (40–150)] and the patient’s loss of wages [INR 200 (150–400)] mainly accounted for the direct non-medical and indirect costs respectively. The median expenditure on smokeless and smoking forms of tobacco was INR 6000 (5400–7200) and INR 2400 (1800–3600) respectively. On multiple linear regression analysis, rural residence, belonging to below poverty line family, being a sole earner in the family, number of months since diagnosis and first visit at a private provider were found to be the significant predictors of total costs of OPMD. Financial protection mechanisms are needed for covering the direct non-medical and indirect costs. Early management of OPMD might mitigate the costs of OPMD.The online version contains supplementary material available at 10.1186/s13104-021-05814-2. Oral cancer is almost always preceded by clinically visible lesions that are noncancerous, to begin with, and which have therefore been termed oral potentially malignant disorders (OPMD) , 2. OralExisting literature on costs of OPMD, none from India, focused on the costs borne by the healthcare facilities in treating these patients , 9. The We conducted a cross-sectional study in the outpatient department (OPD) of the Otorhinolaryngology Department of Sir Takhtsinhji Hospital of Bhavnagar district of Gujarat from July–September 2019. The population of Bhavnagar district and Bhavnagar city as per census 2011 is around 2.8 million and 0.6 million respectively  . Patientα2S2/W2, Table . The medAge in years, male gender, rural residence, being a sole earner in the family, belonging to below poverty line family, number of months since diagnosis, number of years since consuming smokeless tobacco, and having the first visit at a private provider were the variables found to have a p-value < 0.2 on unadjusted uni-variable linear regression analysis . Age in years, male gender, and the number of years since consuming smokeless tobacco were not significantly predicting costs of OPMD.On performing multiple linear regression, rural residence, belonging to below poverty line family, being a sole earner in the family, number of months since diagnosis, and first visit at a private provider were found to be significantly predicting total costs of OPMD Table . Rural rPatients with OPMD incurred lower direct medical and total healthcare costs when compared with their monthly family incomes in this study setting. As might be the case with patients managed at a government hospital, patients with OPMD in the present study incurred lower direct medical costs due to free diagnostic and treatment services. Over 90% of patients first visited our facility (government-run) for the management of OPMD. The low percentage of patients visiting private providers indicates their healthcare-seeking behaviour towards the public health system. Almost half of the patients resided in rural areas where specialist services of the private sector are scarce. Also, since three-fourth of the patients were diagnosed on the first visit when the cost survey took place, the costs for pre-diagnostic visits elsewhere were not incurred. None of the patients required hospitalization and were managed on an OPD basis, cancelling out costs of hospitalization.The loss of wages of patients with OPMD and the travel cost for attending health facilities were the prominent costs reported in this study. Early identification and appropriate case management of OPMD would be pertinent in reducing the costs. The current study found substantially high expenditure on tobacco consumption  among OPMD patients. Studies in India have documented similar expenditures on tobacco consumption in other populations as well –17. TobaThe present study found that rural residence, duration since diagnosis of OPMD, and having the first visit at a private practitioner increased the costs of OPMD. Patients staying in rural areas have to travel to the tertiary care centres, which are usually found in towns or cities, for the care of their OPMD complaints. Also, due to the long travel, there is a possibility of delayed care-seeking and a longer duration of diagnosis since the onset of symptoms. Also, the poor accessibility to specialists in rural areas makes patients seek care at local traditional healers or quacks or private practitioners, thereby adding to the costs incurred for OPMD. These findings were supported by research on the determinants of catastrophic expenditures in patients with cancer –20.We conclude that the costs of OPMD in our setting are low, however, the need for financial protection mechanisms such as universal cash transfers should be explored to overcome the direct non-medical and indirect costs. This is the first study estimating the healthcare costs of patients with OPMD in a sparsely-populated district of western India with a high caseload. Further studies are required in densely-populated urban metropolitan settings of India for estimating costs, and exploring the effects of early management, tobacco cessation, and financial protection on cost mitigation among patients with OPMD.There are a few inherent limitations of studies estimating costs. Since the costs are estimated based on a survey, recall bias is inherent. However, we interviewed more than 90% of patients within a month of diagnosis to avoid recall bias. We could not use any scale for assessing the severity of OPMD among the patients, which might have differentially affected the costs incurred. Also, costs incurred till the resolution of symptoms or cure or progression could not be included as we did not follow up with the patients. Our study did not employ the ‘cost of illness’ perspective, that is, we did not estimate the financial burden incurred by the hospital for the care of the patients. Since patients taking treatment at a private facility were not included, the study findings can be generalized only to patients being treated at government health facilities in similar semi-urban settings of India.Additional file 1. Table showing the unadjusted linear uni-variable regression analysis for variables predicting total costs of OPMD at Bhavnagar during July-September 2019.


Aspergillus and Pseudomonas compete in nature, and are the commonest bacterial and fungal pathogens in some clinical settings, such as the cystic fibrosis lung. Virus infections of fungi occur naturally. Effects on fungal physiology need delineation. A common reference Aspergillus fumigatus strain, long studied in two (of many) laboratories, was found infected with the AfuPmV-1 virus. One isolate was cured of virus, producing a virus-free strain. Virus from the infected strain was purified and used to re-infect three subcultures of the virus-free fungus, producing six fungal strains, otherwise isogenic. They were studied in intermicrobial competition with Pseudomonasaeruginosa. Pseudomonas culture filtrates inhibited forming or preformed Aspergillus biofilm from infected strains to a greater extent, also seen when Pseudomonas volatiles were assayed on Aspergillus. Purified iron-chelating Pseudomonas molecules, known inhibitors of Aspergillus biofilm, reproduced these differences. Iron, a stimulus of Aspergillus, enhanced the virus-free fungus, compared to infected. All infected fungal strains behaved similarly in assays. We show an important consequence of virus infection, a weakening in intermicrobial competition. Viral infection may affect the outcome of bacterial–fungal competition in nature and patients. We suggest that this occurs via alteration in fungal stress responses, the mechanism best delineated here is a result of virus-induced altered Aspergillus iron metabolism. Aspergillus wild-type strains have been known since 1970 [Aspergillus physiology or virulence.Viruses naturally infecting Polymycoviridae was initially reported in 2015 [https://talk.ictvonline.org/; accessed on 5 April 2020). Polymycoviridae currently accommodate one genus Polymycovirus and 10 species, including Aspergillus fumigatus polymycovirus 1. Members of the Polymycoviridae family and related viruses have usually four [The virus family lly four  and up tlly four  dsRNA gelly four , althougPseudomonas–Aspergillus interactions  was cured from AfuPmV-1, using the protein synthesis inhibitor cycloheximide [Aspergillus by protoplast transfection [Isolates: The USA and UK AF293 strains were designated by the California Institute for Medical Research (CIMR) # 10-53 and 18-95, respectively. P. aeruginosa strains [PAO1 and PA14 are extensively studied widely distributed laboratory reference  strains . The usePseudomonas aeruginosa supernatants and molecules on Aspergillus biofilms, as well as sources for materials, were as previously described [In brief, methods used for generating and testing escribed ,16,17,20H)-quinolone), XTT -2H-tetrazolium-5-carboxanilide inner salt), menadione, iron (FeCl3), and RPMI 1640 medium were purchased from Sigma-Aldrich . Trypticase soy agar (TSA) was purchased  and prepared per manufacturer instructions. RPMI 1640 agar was prepared: Briefly, 7.5 g Bacto Agar  in 100 mL distilled water was autoclaved and mixed with 400 mL pre-warmed RPMI-1640 medium.Materials: Pyoverdin, PQS  was incubated in RPMI 1640 medium with or without addition of FeCl3 at 37 °C and 100 rpm for 24 h. Bacterial growth was measured at 600 nm, using a spectrophotometer . Bacterial cultures were centrifuged at 200× g for 30 min, at room temperature, and filtered for sterility (0.22 micron).Methods: eviously . BrieflyPseudomonas effects on Aspergillus forming or preformed biofilms forming biofilm assay: A. fumigatus conidia (2.5 × 104/mL) was seeded in test substances in RPMI 1640 medium, in 96-well plates, at 37 °C, overnight (16 h). Preformed biofilm assay: A. fumigatus conidia (2.5 × 104/mL) was seeded in RPMI 1640 medium, in 96-well plates, at 37 °C, for 24 h. The plates were washed with phosphate-buffered saline, and test substances were added. Plates were incubated at 37 °C, overnight (16 h).Plate assays for the determination of All assays were evaluated by XTT metabolic assay ,21. BrieAspergillus growth, by Pseudomonas supernatants or antifungal drugs, were as described [Assays for testing inhibition of planktonic escribed ; Minimumescribed .Pseudomonas volatiles on Aspergillus growth was assessed by cutting away a 3 mm–wide strip, with a scalpel, out of the agar of an 8 cm diameter TSA agar plate, along a diameter, to separate the plate into 2 semicircular noncontiguous agar halves. The halves were inoculated with 10 microliters of a 107/mL suspension, in RPMI1640 of Pseudomonas, or of Aspergillus conidia, and this co-culture incubated for 72 h, at 37 °C, and then the Aspergillus colony area (πr2) was measured. Each experiment involved 3 replicates.As in the manner of previous studies , the efft-test if two groups were compared, and one-way analysis of variance (ANOVA) combined with Tukey’s post-test for multiple comparisons. All data are expressed as the mean and standard deviation. Data reported as percentages of the control value were compared with Student’s t-test after arcsin transformation of the proportions; these data are presented as percentages. Assays used 4–8 replicate wells for each group studied, for statistical purposes.Statistical analysis: Results were analyzed with Student’s A. fumigatus physiology in the context of the assays to be performed: To assist in interpretation of possible P. aeruginosa effects on A. fumigatus in the assays selected to test such effects, we needed to assess A. fumigatus function in the absence of P. aeruginosa, which is the A. fumigatus control we would use for assessing P. aeruginosa effects. Seven experiments assessing A. fumigatus biofilm metabolism were performed in the milieu to be used for later testing P. aeruginosa effects: RPMI 1640, in the plates, temperature, etc., described. There were 5 to 17 replicates for each of the three A. fumigatus strains  under study, in each of the seven experiments . The mean ± SD for the XTT assays, A490, for the seven experiments was 0.359 ± 0.12, 0.387 ± 0.13, and 0.455 ± 0.14, for the three isolates, respectively. None of these is significantly different from the others, p > 0.05.Effect of virus infection on P. aeruginosa effects (below) involved effects of volatiles on A. fumigatus growth on TSA agar. As will be reiterated, there were, again, not any differences found in the A. fumigatus controls (absence of P. aeruginosa) under the same culture conditions, as described for those assays.Aside from metabolism (assessed by XTT), the other assay of Pseudomonas planktonic supernatant on preformed Aspergillus biofilm: In the first study with preformed Aspergillus biofilm , and P. aeruginosa PAO1 planktonic supernatants, we found the virus-free preformed biofilm resistant to Pseudomonas supernatant  supernatant on biofilm formation  by A. fumigatus was studied (Pseudomonas supernatant. The virus-free Aspergillus is significantly inhibited less.Effect of  studied . All isop < 0.001). At a 1:500 dilution, all three strains were inhibited ≤50% of controls, but the virus-free was inhibited significantly less than either two infected strains . At a 1:1000 dilution, none of the three Aspergillus strains was inhibited.A second experiment of this type was performed with PA14 supernatant. PA14 supernatants proved to be much more inhibitory in these experiments with biofilm formation than PAO1 supernatants, consistent with the reported greater virulence of PA14 compared to PAO1 , owing tAspergillus strain pyoverdine is a Pseudomonas siderophores that is inhibitory to Aspergillus [Aspergillus biofilm (XTT assay) (Aspergillus was significantly less inhibited by pyoverdine. A similar difference was noted with 12.5 micromolar pyoverdine (not shown); the two infected Aspergillus strains were inhibited more at this higher concentration, and p < 0.001 compared to the virus-free.The effect of pyoverdine on ergillus . We compT assay) . The virAspergillus biofilm: PQS is an important Pseudomonas exoproduct that is involved in quorum sensing and the production of virulence factors, and it also has iron-binding properties. It inhibits Aspergillus [Aspergillus biofilm metabolism (Effect of PQS on preformed ergillus  under irtabolism . At everPseudomonas volatiles on Aspergillus strains with the assay method and media described (see Methods): In studies with many Pseudomonas laboratory strains and clinical isolates, and several Aspergillus isolates, we find Pseudomonas volatiles to be inhibitory to Aspergillus [Effect of ergillus . The entAspergillus growth on agar in the presence of Pseudomonas growth on the same plate, as described. In the absence of Pseudomonas, the virus-free and the two virus-infected strains grew equally well on TSA agar. We found that the virus-free Aspergillus was inhibited significantly less by Pseudomonas than the infected (We here tested infected . The expAspergillus strains: Iron stimulates Aspergillus biofilm growth [3 on virus-free vs. virus-infected Aspergillus preformed biofilm metabolism (XTT assay), over a range of iron concentrations  for the virus-free and virus-infected Aspergillus strains were, for amphotericin B, 1 and >8, respectively; for caspofungin, 12.5 and >50; and for voriconazole, ≤0.5 and >8.Drug susceptibility: We compared the susceptibility of virus-free vs. virus-infected Aspergillus, including members of the established families Chrysoviridae, Narnaviridae, Partitiviridae, Polymycoviridae, and Totiviridae [Metaviridae and Pseudoviridae. The viruses tend to be latent, persist, and are difficult to eliminate [A. foetidus mycovirus complex [Candida albicans [Fungal viruses have been described in 22 viral taxa, with five in the genus iviridae . It has iviridae ,28. Mostiviridae ,29,30. Tiviridae . Most ariviridae ,30,31. Eiviridae  and traniviridae  for the iviridae . Most myliminate ,30. Multliminate , for exa complex ,36,37. W complex ,29,38. Talbicans ,28.Saccharomyces cerevisiae, mycoviruses encode protein toxins that kill other fungi, in some by inhibiting glucan synthesis [A. flavus [Cryphonectria parasitica, the fungus causing chestnut blight [All known mycovirus genomes encode for replication enzymes, such as RNA-dependent RNA polymerase. Conversely, not all encode capsid polypeptides or are enclosed in traditional, spherical, or filamentous, protein capsids. This is the case for polymycoviruses and related viruses ,4, that ynthesis ,39 or DNynthesis . Some fuynthesis , such as. flavus . Fungal . flavus , and virt blight . There ht blight .Aspergillus, ~10–50% of isolates in a species have been described as infected [A. fumigatus clinical isolates are virus-infected [Aspergillus viruses can produce infections with only naked dsRNA [In infected . Seven tinfected ,42. The ed dsRNA .Aspergillus that some viruses may increase virulence [Aspergillus viral taxa have no effect on antifungal drug susceptibility [Whereas the preponderance of evidence associates non-latent virus infection with fungal hypovirulence, as is the case for a variant of AfuPmV-1 that has one additional dsRNA element and was tested in a murine model , there iirulence ,44, or hirulence . Other stibility , as was Aspergillus virus alters A. fumigatus phenotype. There is evidence for an active competition between Aspergillus and Pseudomonas, a competition that may occur in soil and water. As these two microbes are the most common fungal and bacterial pathogens in many patient groups, the outcome of the competition could be very important for the long-term welfare in the patients  may not apply to all other growth conditions; other growth conditions would need to be studied individually. Indeed, as yet unpublished studies, with a related virus, in an entomopathogenic fungus, indicate differences between that infected and uninfected fungus, with respect to growth, are medium-dependent [The differences demonstrated in susceptibility to ependent .Pseudomonas products are linked to a virus-induced alteration in Aspergillus iron metabolism; iron metabolism is key to Aspergillus physiology, as recently reviewed [Aspergillus strains were discovered, which would explain the strain differences in response to iron-denying Pseudomonas products . It is also probable that virus infection weakens Aspergillus’ ability to respond to any stresses, of which iron denial is one. The best evidence for that hypothesis is the differential susceptibility to Pseudomonas volatiles, since small lipophilic organic molecules appear to be the mechanism of such inhibition [The differential effect of virus on iron-stimulated fungal growth, particularly when coupled with the findings of effects of iron-chelating molecules, such as pyoverdine and PQS, may be a clue that at least some of the resistance differences to reviewed . Subsequhibition .Aspergillus, to another microbe, Pseudomonas, coupled with studies indicating fungal virulence may be depressed by viral infection, and that some viruses may lyse fungi [Our novel observations about altered susceptibility of a fungus, se fungi ,28,29,38se fungi ,31, anal


VTH, ΔION, Δgm, and ΔSS. The simulation analysis also shows a simpler recovery mechanism in this case.The recent corona outbreak has necessitated the development of a label-free, highly sensitive, fast, accurate, and cost-effective biosensor for the detection of SARS-CoV-2 virus. This study records the label-free electrical detection of the SARS-CoV-2 virus using the gate-all-around junctionless field effect transistor (GAA-JLFET) that detects the virus because of the electrical properties (dielectric constant and charge) of spike protein, envelope protein, and virus DNA, for a highly sensitive and real-time bio-sensor. GAA-JLFETs are suitable for this application because of their highest gate controllability, potential vertical stacking, current industry trend compatibility, inherent ease of fabrication, and higher sensitivity. The SARS-CoV-2 virus is first immobilized in the etched nano-cavity embedded beneath the gate electrode, which is then used to detect it. The SARS-CoV-2 virus detection has been calibrated based on the change in system electrical properties after virus immobilization. For effective virus detection, the work takes into account both the dielectric property of S protein and the charge of DNA at the same time. The sensitivity has been calculated using Δ The present work investigates the label-free electrical detection of the SARS-CoV-2 virus using GAA-JLFET that detects the spike protein, envelope protein, and virus DNA, for a highly sensitive and real-time bio-sensor. Spike protein (S-protein) protrusions and deposits on the envelope protein (E-protein) and membrane protein (M-protein). A nucleocapsid protein (N-protein) surrounds the RNA, the variation in genome sequence is termed as mutation. It can be the result of genetic copying errors during replication of organisms because of exposure to certain chemicals, ionizing radiations, etc. Mutations cause new variations in a species, and cumulative mutations can even lead to the creation of newer species. These mutations could result in virus strains that are more harmful because they are more contagious. The genome of SARS-CoV-2 is made up of more than 30 000 units of ribonucleic acid (RNA). Nucleotides are the building blocks of DNA. Coronaviruses have the largest genome of all the RNA virus families. SARS-CoV-2 virus detection is primarily achieved by two methods. The first is a real-time reverse transcription-Polymerase Chain Reaction (rRT-PCR),7–10 and the second is an IgG antibody test using Enzyme Linked Immunosorbent Assays (ELISA). ELISA is a typical procedure for virus detection in which the virus's antibody is found in the patient's blood serum instead of the virus itself.11 The second method is based on the rRT-PCR method, in which the kit detects the E-protein gene, RNA (N-protein), open frame reading b1 (OFRb1), and OFRb2 genes in the internal nasal swab.8 Viruses mutate while transmitting from one medium to another. Various SARS CoV-2 virus mutant have been identified and labeled by WHO as B.1.1.7 (Alpha), B.1.351 (Beta), B.1.617.2 (Delta), P.1 (Gamma), B.1.526 (Iota), B.1.427 (Epsilon), B.1.429 (Epsilon), B.1.617 . These mutants lead to various repercussions such as high transmission rate, increased morbidity, high number of deaths, mutants are able to bypass the diagnostic tests and are not detected, less susceptible to neutralising antibodies, whether therapeutic or laboratory experiments, mutants are able to bypass natural antibody and causes reinfections, mutants are able to infect even after vaccination, it also increases certain risk such as multi-organ inflammatory syndrome or long COVID, higher affinity to distinct groups, such as children or individuals with compromised immunity.12This COVID-19 pandemic has wreaked havoc on society and caused global hysteria. Different zoonotic outbreaks such as Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) in 2002–2003  and low microwave power (200 W).18–21This work reports the label-free electrical detection of the COVID-virus based on S-protein (dielectric constant) and C-DNA (charge density) by deploying the gate-all-around junctionless field effect transistor (GAA-JLFET). Here highly sensitive and real-time bio-sensors are designed. GAA-JLFETs are the most suitable for this application because of their highest gate controllability, potential vertical stacking, current industry trend compatibility, inherent ease of fabrication, and higher sensitivity. The SARS-CoV-2 virus is first immobilized in an engraved nano-cavity embedded beneath the gate electrode, and then it is used to detect it. The identification of the SARS-CoV-2 virus is now calibrated based on the change in system electrical properties following virus immobilization. The study takes into account both the dielectric property of S protein and the charge on the DNA molecule simultaneously for reliable virus detection. The GAA-JLFET structure can be realized using the process suggested in.The following is the organization of the work: the device structural parameters, materials, and doping levels are discussed in Section 2. The effect of cavity thickness variation on bio-sensor sensitivity, as well as sensitivity variation with dielectric modulation and charge density modulation is investigated in Section 3. Section 4 brings the research to a conclusion.2x–y plane, z–y plane, z–x plane are shown in CNC shows the capacitance of the nano-cavity, where Cox stands for the capacitance of the oxide layer, and where CDep shows the depletion capacitance of the device. The capacitance model of the device illustrates that the variation in the electrical property of the virus immobilized in the cavity will vary the capacitance of the nano-cavity and this will lead to the change in the device characteristics sensing the virus. The gate length is kept in accordance with the state-of-art technology i.e. 45 nm with work-function of gate electrode considered as 5.93 eV . All the device structural parameters are shown in 18 cm−3 of n-type. The cavity region is 15 nm thick under three gates for the immobilization of bio-molecules. Bio-molecule adsorption test is performed during fabrication of sensor by keeping bio-molecule in the cavity till equilibrium between the adsorbed bio-molecule and the bio-molecule in the cavity is reached. A uniform temperature of 25 °C is kept using a thermostat shaker bath. The concentration of bio-molecule present in the solution before and after adsorption is used to calculate the amount of bio-molecule adsorbed. Further, a 5 nm thick SiO2 layer is used for the enhanced binding of the bio-molecules. The study is done by deploying technology computer aided design (TCAD) tool Silvaco ATLAS.22 The drift-diffusion transport model (cvt) is used to model the carrier transport. Concentration dependent and field dependent mobility models are used along with SRH and auger recombination for the precise modeling of carrier recombination. Band gap narrowing model (BGN) is also used. Gummel and newton trap numerical solvers have been used to improve the convergence. Here, the basic bio-molecule detection mechanism is based on the dielectric and charge based modulation effect due to the SARS-CoV-2 virus immobilization in the nano-cavity embedded below the gate electrode.23–27Gate-all-around Junctionless FET (GAA-JLFET) based SARS-CoV-2 virus sensor has been designed because of its single uniform doping, thus easy fabrication and cost-effectiveness. The 3D structure of the GAA-JLFET sensor, 2D cross-sectional view along the 313–15,28–31 This property can be utilized for the electrical detection of the virus. Further, the presence of charge density in the DNA of the bio-molecule also provides scope for virus detection based on charge density. In this work, we have studied the detection of viruses based on dielectric modulation as well as charge modulation.32–36The dielectric constant of protein protrusions of SARS-CoV-2 virus varies between 2 to 4.3.1x–z cross-section for the dielectric constant of bio-molecule in the cavity is taken as 1 (air), 2 and 4 for VGS = 1 V and VDS = 1 V. x–z cross-section for the dielectric constant of bio-molecule in cavity is taken as 1 (air), 2 and 4 for VGS = 2 V and VDS = 1 V and x–z cross-section for the dielectric constant of bio-molecule in the cavity is taken as 1 (air), 2, and 4 for VGS = 3 V and VDS = 1 V. It is observed from the contour plot in VGS = 1 V leading to a decrease in drain current and increase in threshold voltage with an increase in the dielectric constant. The decrease in surface potential with an increase in dielectric constant is minimized for VGS = 2 V thus the decrease in drain current with an increase in dielectric constant reduces. Further, the surface potential increases with an increase in dielectric constant for VGS = 3 V leading to increase in drain current with increase in dielectric constant. This drain current behaviour with respect to gate voltage/transfer characteristics are shown in VTH, ION, ION/IOFF, gm and SSavg of GAA-JLFET sensor with the dielectric constant variation of bio-molecule are listed in VTH, ΔION, and Δgm are shown in VTH, ΔION and Δgm represent the change in VTH, ION and gm with respect to air in the cavity, respectively and is given asIn our analysis, we have taken the dielectric constant of SARS-CoV-2 virus as 2 and 4. avg of the sensor also varies with the dielectric constant variation of the bio-molecules. The SSavg of the sensor improves with an increase in the dielectric constant of bio-molecule. Here, SSavg is defined asThe SSavg plot is shown in VTH is in the range of 0.13 V and 0.24 V, ΔION is in the range of 10−6 A and Δgm is in the range of 10−6 A/V, also the variation of SSavg with variation in dielectric constant ensures good sensitivity with dielectric modulation and thus precise detection of bio-molecule/virus.The SS3.212 cm−2 to 1 × 1012 cm−2 with a fixed dielectric constant of bio-molecule, the dielectric constant is 1.The presence of charge density in the DNA of the virus makes it traceable with charge density modulation. The interface trap charge density considered for the study is − 1 × 10x–z cross-section with bio-molecule charge density taken as 1 × 1011 cm−2, 5 × 1011 cm−2, and 1 × 1012 cm−2, respectively at VGS = 3 V and VDS = 1 V. The flat band voltage decreases with an increase in positive charge  and thus it leads to an increase in the effective potential .37 The potential contour shows an increase in potential with increase in the positive charge density of bio-molecules. The increase in surface potential with an increase in the positive charge density leads to an increase in the drain current and thus a decrease in threshold voltage as shown in VGS for peak transconductance with an increase in positive charge density. The increase in the positive charge density leads to a decrease in VGS value for peak transconductance. The VTH, ION, ION/IOFF, gm, and SSavg of GAA-JLFET sensor with the positive charge density variation of bio-molecule is listed in VTH and ΔION is shown in VTH and ΔION denote the change in VTH and ION with respect to the neutral bio-molecule in cavity, respectively and is given asavg of the sensor also varies with the positive charge density variation of bio-molecule. The SSavg of the sensor degrades with an increase in the positive charge density of bio-molecule. The SSavg plot is shown in VTH is in the range of 1–2 V and ΔION is in the range of 10−5 A, also the variation of SSavg with variation in positive charge density ensures good sensitivity with positive charge density and thus precise detection of virus.The SSx–z cross-section with bio-molecule charge density is considered as −1 × 1011 cm−2, −5 × 1011 cm−2, and −1 × 1012 cm−2 respectively at VGS = 3 V and VDS = 1 V. The flat band voltage increases with an increase in negative charge and thus it leads to decrease in effective potential.37 The potential contour shows a decrease in potential with an increase in negative charge density of the bio-molecules. The decrease in surface potential with an increase in the negative charge density leads to a decrease in the drain current and thus an increase in threshold voltage as shown in VGS for peak transconductance with an increase in negative charge density. The increase in negative charge density leads to an increase in VGS value for peak transconductance. The VTH, ION, ION/IOFF, gm, and SSavg of GAA-JLFET sensor with the negative charge density variation of bio-molecule are listed in VTH and ΔION are shown in VTH and ΔION represent the change in VTH and ION with respect to neutral bio-molecule in the cavity, respectively and as given by avg of the sensor also varies with the negative charge density variation of bio-molecule. The SSavg of the sensor improves with an increase in negative charge density of bio-molecule. The SSavg plot is shown in VTH is in the range of 1–2 V and ΔION in the range of 10−5 A, also the variation of SSavg with variation in negative charge density ensures good sensitivity with negative charge density and thus precise detection of virus.4VTH, ΔION, Δgm and SS. This higher variation in these electrical parameters helps in realizing highly sensitive biosensors. GAA-JLFET is deployed here owing to its easy fabrication and higher sensitivity due to higher gate controllability with the increased number of gates. The basic mechanism for SARS-CoV-2 virus detection is the immobilization of the virus in the etched nano-cavity embedded beneath the gate electrode. Further, the change in the device's electrical properties post virus immobilization is calibrated for the SARS-CoV-2 virus detection. This study shows very high sensitivity towards DNA charge density. The TCAD investigation pledges for the potentials of this structure for the array-based screening and in vivo bio-species diagnostics.This work reports the label-free electrical detection of SARS-CoV-2 virus using the gate-all-around junctionless field effect transistor (GAA-JLFET). The study takes into account simultaneously both the dielectric property of S protein and the charge of DNA for detection of the SARS-CoV-2 virus. The sensitivity has been analyzed in terms of ΔThere are no conflicts to declare.RA-012-D1RA08587E-s001


P ≤ 0·05) reduced in the presence of magnesium carbonate, calcium carbonate and the chalk blend; the infectivity was reduced by ≥2·29 log10 50% tissue culture infective dose (TCID50) immediately upon on contact and by ≥2·46 log10 TCID50 within 1 h of contact. These findings suggest that the infectivity of coronaviruses is reduced by climbing chalk, limiting the risk of potential fomite transmission.There does not appear to be any studies in the published literature on the stability of SARS‐CoV‐2 in climbing chalk powder , which has been hypothesized to pose a potential risk of fomite transmission of coronavirus disease 2019 (COVID‐19) within climbing gyms. The aim of this study was to determine the infectivity of a model human coronavirus HCoV‐OC43 in the presence of climbing chalk powder on a dry plastic surface. The stability of HCoV‐OC43 on a plastic surface dusted with climbing chalk powders  was determined by titration on BHK‐21 fibroblast cells. No chalk and no virus controls were included. HCoV‐OC43 was stable on the plastic surface for 48 h. The stability of HCoV‐OC43 was significantly ( The emergence of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) and subsequent development of the coronavirus disease 2019 (COVID‐19) pandemic has led to the widespread implementation and promotion of public health interventions, such as social distancing, closure of nonessential businesses, enhanced disinfection and handwashing to limit transmission of COVID‐19  and has a similar overall structure; both possess a single‐stranded positive‐sense RNA genome and are enclosed in a lipid envelope with spike proteins protruding from the surface , stainless steel (5·6 vs 4·2 h) and plastic (6·8 vs 7·6 h).In this study, human coronavirus (HCoV‐OC43), a common causative agent of upper respiratory tract infections in humans  in the presence of climbing chalk powder on a dry plastic surface.10 50% tissue culture infectious dose (TCID50) sample−1 infectious virus being detected on the surface after 1 h and ≤2·68 ± 0·35 after 24 h (Table P ≤ 0·05) decrease in infectious viral titre was observed until 48 h incubation, where no infectious virus was detected , and after 48–72 h, only 0·5–1·0 log10 ml−1 was detected. No infectious virus was recovered from the no virus controls.HCoV‐OC43 remained relatively stable on the plastic surface without chalk, with 3·74 loge et al.  recovereP ≤ 0·05) reduced by magnesium carbonate, calcium carbonate and the blended chalk  number of viral RNA was present in the magnesium carbonate supernatant compared to the pellet, where 80·3 ± 2·4% of the total RNA was recovered in the supernatant compared to the pellet. There was no significant difference (P > 0·05) in the recovery of HCoV‐OC43 between calcium carbonate and blended chalk supernatants and pellets, with 50·5 ± 16·4% and 39·4 ± 9·7% of the total viral RNA being recovered in the respective supernatants compared to the pellets. Although a proportion of the viral RNA was lost within the chalk pellet, removal of excess chalk could not be avoided due to cytotoxicity of the chalk against the mammalian cell lines. These results do not change the observations of an overall loss in infectivity of HCoV‐OC43 on exposure to the chalk; theoretically if virus from the pellet was also recovered, the maximum increase in infectivity in the blended chalk condition at 0 min would be to 2·9 log10 TCID50 per sample (from ≤2·5 log10 TCID50 per sample) assuming 39·4% recovery, which still results in approximately a 2 log10 reduction. It is important to note that detection of nucleic acids does not distinguish between infective and inactive virus and therefore it was not possible to ascertain the TCID50 sample−1 of HCoV‐OC43 within the pellet. Moreover the behaviour of the virus within the pellet would also be in line with those in the supernatant and therefore the majority are likely be noninfectious.In this study, infectious virus was recovered from the chalk by resuspension in PBS and centrifuging to remove excess chalk. This method was employed to minimize the cytotoxicity of the chalk and allow syringe filtration of the viral supernatant to avoid bacterial contamination. In order to validate the recovery method, the distribution of viral RNA between the supernatant and the chalk pellet after 0 min contact were quantified by quantitative reverse transcription PCR (RT‐qPCR). A significantly greater (10 TCID50 per sample) up to 30 min contact with magnesium carbonate and the chalk blend. No infectious virus was detected after 1 h in the presence of chalk, indicating that ≤2·33 log10 TCID50 infectious virus was present. The infectious viral titre did not change significantly (P > 0·05) over the course of 1 h upon contact with all three chalks. One limitation of this study is the relatively low sensitivity of the infectivity assay, where the limit of detection was 2·33 log10 TCID50 per sample. This can be attributed to cytotoxicity of the chalk supernatants upon BHK‐21 cells, thus requiring a greater level of dilution to quantify infectious virus.The number of infectious virus remained close to or below the detection limit (2·33 log−1 suspension) was previously reported to inhibit the growth of Staphylococcus epidermidis, which was in part attributed to its alkalinity  is in line with that of similar studies measuring the environmental stability of coronaviruses  culture medium supplemented with 10% fetal bovine serum , 100 IU ml−1 penicillin and 100 μg ml−1 streptomycin . BHK‐21 fibroblast cells  were cultured in Dulbecco's Modified Eagle Medium  supplemented with 10% FBS, 100 IU ml−1 penicillin and 100 μg ml−1 streptomycin at 37°C with 5% CO2.HCT‐8 epithelial cells (ECACC 90032006) were cultured at 37°C with 5% CO−1 penicillin and 100 μg ml−1 streptomycin for 7 days at 33°C. HCoV‐OC43 was harvested by aspirating the RPMI‐1640 culture media and centrifuging  to remove cell debris. The virus stocks were stored at −80°C before use. All experiments were performed within a Biosafety Level 2 laboratory.Human coronavirus HCoV‐OC43 (ATCC VR‐1558) was cultured in HCT‐8 epithelial cells in RPMI‐1640 with 5% FBS, 100 IU ml2 PVC PUR plastic swatches.A layer of either magnesium carbonate chalk , calcium carbonate chalk  or a chalk blend  was deposited onto 25 cmμl of the model human coronavirus HCoV‐OC43 (4·79 log10 TCID50 per sample) was transferred via 10 μl droplets to the surface of the chalk and incubated at room temperature for 0, 1, 5, 10, 30 and 60 min. No virus  and no chalk (200 μl HCoV‐OC43) controls were included.Aliquots of 200 g for 10 min to remove excess chalk. The supernatant was syringe filtered , serially diluted in DMEM supplemented with 5% FBS, 100 IU ml−1 penicillin and 100 μg ml−1 streptomycin and plated onto BHK‐21 fibroblast cells seeded in a 96‐well format. The plates were incubated at 33°C with 5% CO2 for four d before the cells were scored for cytopathic effect. The number of infectious viral particles present on each surface sample (log10 TCID50 per sample) was then determined according to the Karber method .Infectious virus was recovered in 3 ml phosphate buffered saline  using cotton swabs, vortexed for 30 s and centrifuged at 2500μl HCoV‐OC43, immediately recovered in three ml PBS as described above and the supernatants harvested. The chalk pellet was resuspended in a further three ml PBS and three cycles of vortexing for 30 s and centrifuging at 2500g for 10 min were performed prior to recovering the supernatant. Resulting supernatants were syringe filtered prior to RNA extraction using the Meridian Bioscience  ISOLATE II RNA Mini Kit according to the manufacturers’ instructions. A no chalk control was included.Chalk powders were inoculated with 200 Total RNA samples were DNase‐treated using the Promega  RQ1 RNase‐free DNase kit prior to reverse transcription using random primers and the GoScript™ reverse transcription system  according to the manufacturers’ instructions. No reverse transcriptase controls were included, where nuclease free water was included in the reaction mix in place of reverse transcriptase.et al. (10 TCID50 per ml).RT‐qPCR was performed manually using the Meridian Bioscience SensiFAST™ Probe Lo‐ROX Kit with HCoV‐OC43‐specific primers  and probe ([6FAM]TGACATTGTCGATCGGGACCCAAGTA[TAM]) as described by Owczarek et al. . No tempn = 3). RT‐qPCR was conducted with technical triplicates of at least two independent experiments (n = 2–3). Distribution of the data was ascertained using the Shapiro–Wilk test (Shapiro and Wilk P ≤ 0·05) in recovered infectious virus (log10 TCID50 per sample) for each condition were determined by the independent samples Kruskal–Wallis test . Significant differences (P ≤ 0·05) in viral nucleic acid recovered from chalk supernatants and pellets was determined using the related‐samples Wilcoxon signed rank test (Wilcoxon Viral infectivity experiments were conducted in triplicate on separate occasions (Wilcoxon .This investigation was funded by The Warehouse Climbing Centre Gloucester, Lakeland Climbing Centre and Association of British Climbing Walls.No conflict of interest declared.


Sir,The SARS-CoV-2 virus which has spread globally and now claimed the lives of 4.5 million people, may be mutating to become more virulent and thus should be a growing concern for both policy makers and individuals alike. There is compelling evidence which suggests that the newer variants of the virus may be spreading more via fine aerosol particles as compared to their predecessors .The mechanism of transmission of the virus has been a topic of intense debate among physicians, as previously it was understood that the initial variants of the virus were generally isolated to heavier droplets which would sink and settle on a surface fairly rapidly and thus be spread directly through contact, other routes of transmission via the faeco-oral route have also been established. Aerosol transmission of the virus has remained a somewhat of a grey area; however new-found research and studies conducted on the newer variants of the SARS-CoV-2 virus indicate that the aerosol route of transmission is being utilized [Aerosols can be produced from various bodily excreta and secretions which contain the SARS-CoV-2 virus. These secretions and excreta which contain the virus can be aerosolized into infectious particles in a multitude of mechanisms. Various secretions chiefly those such as respiratory secretions are aerosolized via menial everyday tasks such as sneezing, talking, exhaling and coughing. Various surgical and interventional procedures such as non-invasive ventilation, tracheal intubation and bronchoscopies also provide the necessary mechanical energy to form aerosolized particles and thus increase the risk to medical personnel performing such procedures. SuperaddStudies conducted at the University of Maryland have concluded that individuals who were infected with the alpha variant of the virus carried up to eighteen times the amount of viral RNA in the aerosols they produced as compared to the older and less virulent variants. The load of viral RNA in these aerosols being high enough to culture the virus from the air samples taken in the laboratory. This evidently relating to a more virulent strain, depicting how the virus has mutated to increase its repertoire of transmission .Supplementary evidence from studies undertaken in Singapore which compared fine (≤5μm) and coarse (>5μm) respiratory aerosols produced when breathing, talking, and singing concluded that the fine aerosols contained a greater amount of RNA viral copies than the coarse aerosols. The importance of this finding is rooted in pathophysiology; as particles less than 6μm in size are able to overcome the respiratory systems natural defences and thus can reach the deeper segments and lobes of the lungs. An alarming factor is that the buoyancy of these fine aerosols means that the virus can remain airborne for a prolonged period after being aerosolized and thus adds a new dimension to the risk of infection in poorly ventilated spaces. Due to the buoyancy of these fine aerosolized particles, the capability of them to remain airborne for extended periods increases the likelihood of inoculation into a desired host . These fine aerosolised particles therefore drastically increase the risk to those individuals not wearing masks .Greenhalgh T et. al, has published 10 scientific reasons in the support of the airborne transmission of COVID-19. 1) Superspreading events account for a substantial number of COVID-19 infections. On analysis of human behaviour in such events it has been concluded that long-range transmission of the virus has taken place in settings where the production of aerosolized particles is maximal, thus supporting the notion of airborne transmission. 2) The long range transmission of the virus to people in separate rooms in quarantine hotels has been recorded. 3) Asymptomatic and pre-symptomatic transmission account for roughly one third of the cases globally where individuals spread the virus through speaking, singing etc. 4) Transmission of the virus is higher indoors than outdoors. 5) Nosocomial infections have been recorded where strict protocol and use of PPE for droplet, but not aerosolized infection has been adhered to. 6) Viable SARS-CoV-2 has been isolated in air samples from bedrooms occupied by COVID-19 patients, where no invasive or particulate generating procedure has been performed.7) The SARS-CoV-2 virus has been detected in building ducts and hospital filters which can only be reached via aerosolized particulate matter. 8) Studies involving infected animals, with interconnecting ducts systems to cages with healthy animals have shown the transmission of the virus which can thus only be logically explained via aerosols. 9) No study has provided strong evidence to consistently refute aerosol transmission. 10) Limited evidence is available to support other predominant routes of infection. The assumption that transmission through close physical contact and proximity is flawed. Large respiratory droplets or fomites have historically (for decades) been used to deny the airborne transmission of tuberculosis and measles which has subsequently been proven otherwise [As of May 18, 2021 the WHO officially updated and recognized aerosol transmission as one of the routes used by the virus to infects its host, this was instituted after over 200 experts wrote an open letter to the WHO insisting that the evidence of airborne transmission of the virus particularly through aerosols had to be recognized .The newly found evidence and acceptance of fine aerosol transmission in the newer variants of the SARS-CoV-2 virus should be a growing concern for both policy makers and individuals alike. By virtue of the fact that the virus spreads through such means, increases the risk of transmission and contamination highly as the virus can be transmitted via long range and through common ventilation and duct systems. In light of this it is now pertinent for legislation to support the use of personal protection equipment to better safeguard the health of the public.Indrajit Banerjee, Sir Seewoosagur Ramgoolam Medical College, Belle Rive, MauritiusJared Robinson, Sir Seewoosagur Ramgoolam Medical College, Belle Rive, MauritiusDated the 20 September 2021


The genome contains the conserved nonanucleotide sequence, TAATATTAC and other sequence features typical of the family Geminiviridae, and encodes two and four open reading frames in the virion-sense and the complementary-sense strands, respectively. Genome-wide pairwise identity analysis revealed that the novel virus shares less than 65.6% identity with previously characterized geminiviruses. Phylogenetic and recombination analysis indicated that this virus was placed in a unique taxon within the family Geminiviridae and potentially arose from recombination. An infectious clone of this virus was further constructed and its infectivity was tested in different species of plants. Successful infection and characteristic symptoms were observed in Glycine max, Nicotiana benthamiana, N. tabacum, N. glutinosa, and N. tabacum cv. Samsun plants. Taken together, this virus represents a member of an unclassified genus of the family Geminiviridae, for which the name soybean yellow leaf curl virus is proposed.Soybean is a major legume crop that plays an important role in food production, industrial production, and animal husbandry. Here, we characterize a novel soybean-infecting monopartite geminivirus identified in China. Analysis of the contigs  Geminiviridae is a family of circular single-stranded DNA (ssDNA) viruses that constitute the largest family of known plant-infecting viruses and have caused many devastating diseases in a wide range of crops, vegetables, and ornamental plants worldwide  (ToLCJV) (accession number AB162141.1), and the other region of about 1600 nt has no significant similarity with known sequences. Pairwise identity of the 2782 bp sequence with the complete genome sequence of ToLCJV revealed that they share 65.6% genome-wide pairwise identity, supporting the likelihood that the circular DNA isolated from the symptomatic soybean leaves represents a previously uncharacterized geminivirus.To identify possible viruses in the collected soybean samples, total RNA was extracted from symptomatic leaves and subjected to deep sequencing using the Illumina Hiseq2000 platform. A total of 20,380,853 clean reads were obtained after filtering the 20,914,978 raw reads by trimming low-quality reads, 5′- and 3′-adapter sequences, and poly A/T/C/G reads. To determine whether the identified soybean-infecting geminivirus is associated with a putative DNA-B component, an alphasatellite, or a betasatellite, the DNA sample obtained from the same diseased soybean plant was used as a template and PCR was performed using degenerate primer pairs, CR01/CR02 , UNA101/https://www.ncbi.nlm.nih.gov/orffinder/, accessed on 27 November 2019), six recognizable ORFs-encoding proteins composed by >96 amino acids were identified, considering both strands of the SbYLCV genome. The arrangement of the six ORFs within the SbYLCV genome resembles those described for monopartite begomoviruses, topocuviruses, turncurtoviruses, maldoviruses, and opunviruses, with two ORFs (V1 and V2) on the virion strand and four  on the complementary strand  with that of ageratum yellow vein virus. Taking into account the nucleotide sequence divergence of the full-length genome and the amino acid sequence of the predicted proteins encoded by SbYLCV, SbYLCV identified in this study is distinct from currently classified geminiviruses and represents a novel species.BLASTP searches using the amino acid sequences of the six individual ORFs of SbYLCV revealed that most of the predicted SbYLCV proteins are highly divergent from those of known geminiviruses. The predicted V1 protein of SbYLCV shares a 32.37% of amino acid sequence identity with the coat protein (CP) of sweet potato symptomless virus 1. SMART analysis showed that the V1 protein was identified as a geminivirus CP. Although the predicted V2 protein had the highest amino acid sequence identity (41.89%) with the V2 protein of bYLCV V2 . On the bYLCV V2  [23,24,2To understand the evolutionary relationship of SbYLCV to other known geminiviruses, phylogenetic trees were individually constructed using the nucleotide sequences of the full-length genomes and the amino acid sequences of CP and Rep encoded by selected geminiviruses. The full-length genome-based phylogeny showed that SbYLCV does not group with geminiviruses belonging to any of the established 14 genera A. PairwiEragrostis curvula streak virus (ECSV)  B. Compars (ECSV) .When the phylogenetic tree was generated using SbYLCV Rep amino acid sequences together with the Rep of other representative geminiviruses, it was inferred that the SbYLCV Rep sequence groups together with the Rep of begomoviruses , an obsep value, 2.66 × 10−24), GENECONV , Bootscan , MaxChi , Chimera , and SiSscan . The breakpoints for this predicted event ranged from nucleotides 1674 to 2760. The sequence of stachytarpheta leaf curl virus (accession number AJ810157.1) in the analyzed dataset was implicated as the putative minor parent, and those of Dolichos yellow mosaic virus (accession number AM157413) and an unknown virus, resemble the putative major parent , five of the 52 soybean plants inoculated with pBin-SbYLCV-2A displayed leaf curling and yellowing symptoms, which were visually different from that inoculated with the empty vector pBinPLUS  agroinfiltrated with pBin-SbYLCV-2A, but not in those agroinfiltrated with the empty vector pBinPLUS  and pBinPLUS . Southera plants B, indicaSolanaceae crops, the infectivity of SbYLCV was further assessed on four other plant species of the family Solanaceae. Agroinoculated plants were closely monitored for symptom development. At 21 dpi, a typical leaf curling symptom was visible in the systemic leaves of N. tabacum, N. tabacum cv. Samsun, and N. glutinosa plants agroinfiltrated with pBin-SbYLCV-2A  with ToLCJV. These observations suggest that this novel circular DNA virus represents a highly divergent species of a putative new genus of the Geminiviridae family, for which the name soybean yellow leaf curl virus (SbYLCV) is proposed.The arrangement of the six predicted ORFs encoded by the novel circular DNA virus is similar to that of monopartite begomoviruses, topocuviruses, turncurtoviruses, maldoviruses, and opunviruses. It also shares several common features with known geminiviruses, including the stem-loop structure and the conserved nanonucleotide, TAATATTAC, within the intergenic region and several critical motifs in the Rep ORF. However, pairwise sequence identity comparison and phylogenetic analyses of the full-length genome showed that this novel DNA virus is distinct from the established 14 genera of the Sequence analysis of the predicted SbYLCV proteins revealed that the Rep and C4 ORFs are closely related to those of begomoviruses, whereas the predicted V1, V2, C2, and C3 proteins share less than 52% amino acid sequence identity with those of other reported geminiviruses. This indicates that SbYLCV is a recombination-prone virus, which was further supported by six recombination detection methods implemented in the RDP software. Recombination is an important force driving geminivirus evolution, which can provide a strong selective advantage in virus species at the species and genus level. Previously, numerous begomoviruses have been reported to arise from recombination and the resulting recombinants often have an increased fitness or pathogenesis. The acquisition of a more virulent C4 arisen by recombination results in a more virulent begomovirus that does not require a betasatellite for symptom induction . It woulBetasatellites are small circle ssDNA molecules that are frequently found in association with many begomoviruses and occasionally with mastreviruses . In linePrevious studies have shown that some functions of positional homologs such as Rep are conserved across geminivirus species and genera, whereas other geminivirus-encoded proteins may display a diversity of functions in different species . For exaGeminiviridae revealed that SbYLCV forms a distinct clade, but was most closely related to the CP of mastreviruses and ECSV. Mastreviruses are known to be transmitted by leafhoppers. Although the insect vector responsible for SbYLCV transmission in the field is scanty at this stage, the availability of the infectious clone developed here and their successful infection on soybean and other experimental hosts allow us to address this issue in the near future.Insect vectors play significant roles in geminivirus spread and evolution in nature, and CP is the only geminiviral protein that is involved in geminivirus transmission. The specificity between CPs with their insect vectors reflects an evolutionary relationship between geminiviruses and their transmission vector. A phylogenetic analysis of the predicted amino acid sequence of the CP of SbYLCV with the CP of geminiviruses representing all the 14 genera of the family In summary, we have detected and characterized a novel soybean-infecting monopartite geminivirus in China. This novel geminivirus can replicate and establish a successful infection in inoculated soybean plants and five experimental hosts. These findings provide useful tools for the understanding of plant and geminivirus interactions and the selection of resistant soybean cultivars, which will be beneficial to the development of management strategies for disease control.


Given the importance of the spine in carrying out daily movements, adolescent idiopathic scoliosis (AIS) can significantly limit the range of motion (ROM). Severe forms of AIS are treated surgically, most commonly with posterior spinal fusion and instrumentation, which may also reduce spine ROM. This review is the first to describe the literature on total spine ROM in patients with AIS before and after corrective surgery. A systematic literature search was performed using PubMed and Google Scholar to identify articles reporting global spine ROM in AIS patients. Following the guidelines of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA), 486 articles were initially identified. Two independent reviewers (YM and JH) assessed eligibility for inclusion.A total of 11 articles fit the inclusion criteria. AIS in untreated patients seems to limit axial and coronal plane ROM based on the degree of curve severity, with more severe curves having less ROM. More research comparing total spine ROM in untreated AIS patients to that of healthy controls is needed. In those undergoing spinal fusions, the lowest instrumented vertebra and surgical approach appear to minimize further reductions in ROM; however, the findings are mixed. Vertebral body tethering (VBT) shows promising preliminary results in treating AIS while preserving motion; however, long-term outcomes have yet to be assessed for this novel procedure. The results of this systematic review suggest that further research is required before treatment strategies can be modified for surgically treating patients with AIS to take into account the effects of treatment on changes in spine mobility. Scoliosis is a complex three-dimensional spinal deformity with both vertebral rotation and lateral curvature of at least 10° . Genome-The spine is the central axis for truncal rotation about the pelvis, which permits lateral bending and anteroposterior flexion-extension. Daily movements, sport, and physical activities involve complex functioning of the healthy spine in three dimensions. For example, forward flexion and axial rotation for picking up an object, and axial rotation, lateral bending, and forward flexion for throwing a ball fast. The pelvis and joints distal to the spine along the kinetic chain operate within expected excursions and application of joint forces to execute the whole movement. Spinal deformities like AIS restrict or change spine motion and pelvis orientation, thereby impacting distal joint excursions and load bearing to achieve the motion .Given the importance of spine motion on the ability to execute everyday activities, there is a need to study how spinal deformities such as AIS affect spine motion, and more importantly, to see whether corrective surgery restores or further worsens spinal mobility. This is the first review article to describe the literature on total spine range of motion (ROM) in patients with AIS before and after corrective surgery.MethodsA systematic literature search was performed in August 2021 using PubMed and Google Scholar. Customized queries including the following keywords with AND/OR operators were entered into the search engines: AIS, adolescent idiopathic scoliosis, motion, and ROM. The search was limited to full-text original (retrospective or prospective) articles published in English after the year 2000. Papers that evaluated total spine ROM in patients with AIS before and/or after corrective surgery were included. Papers that focused on motion in specific spinal segments instead of total spine ROM were excluded  or focused on motion only in the cervical or lumbar spine. Reference lists were scanned for any additional articles that fit the criteria. A total of 11 articles published between 2002 and 2021 were ultimately included in this review.One paper described total spine ROM before corrective surgery relative to the degree of spine curvature. The other 10 papers studied the change in spine motion in the front, lateral, and/or axial planes following fusion and instrumentation  on spine ROM. Engsberg et al.  evaluateLee et al.  specificUehara et al.  conducteUdoekwere et al.  confirmeOhashi et al.  built upImpact of Length of FusionOne study directly investigated the effect of length of fusion on spine motion. Danielsson et al.  conducteThree studies are the first to investigate the effects of the surgical approaches on functional outcomes. Helenius et al.  comparedSimilarly, Engsberg et al.  carried Recently, Pehlivanoglu et al.  conducteThis review of the current literature on spine ROM in patients with AIS before and after corrective surgery brings to light the many gaps that still need to be filled. We know that curve severity prior to treatment may predict losses in axial and coronal plane ROM and that patients may have left-right lateral bending asymmetry based on the direction of their curve. However, we have yet to take these changes in mobility into account when surgically treating these patients and have thus far placed the greatest emphasis on maximizing curve correction - a metric that has yet to be proven to improve functional outcomes.The literature on postoperative changes in global spine mobility is growing. One study found no correlation between Cobb angle and mobility in any plane and there are mixed findings on the impact of a more distal LIV. Unfortunately, only four studies have compared different surgical approaches based on functional outcomes for patients with AIS. Given the importance of restoring mobility and advancements in the surgical treatment of severe AIS, there is an even greater need to study how new technology and instrumentation may affect mobility.A better understanding of spine mobility in untreated AIS and after operative correction with modern surgical options would help guide treatment decisions, optimal level selection, and pre-surgical discussion to improve the quality of life of patients with AIS.


Gastroretentive drug delivery system (GDDS) are novel systems that have been recently developed for treating stomach diseases. The key function of all GDDS systems is to control the retention time in the stomach. However, research into the bulk density or entanglement of polymers, especially regarding their effects on drug float and release times, is scarce.In this research, we prepared the floating core-shell beads carrying tetracycline. The ratio of chitosan and xanthan gum in the shell layer was changed to modify polymer compactness. Tetracycline was encapsulated in the alginate core.Using scanning electron microscopy (SEM) techniques, we observed that the shell formulation did not change the bead morphology. The cross-sectional images showed that the beads were highly porous. The interaction between anionic xanthan gum and cationic chitosan made the shell layer dense, resisting to the mass transfer in the shell layer. Due to the high mass transfer resistance to water penetration, the longer float and delivery time were caused by the dense surface of the beads. The cell culture demonstrated that floating core-shell beads were biocompatible. Importantly, the beads with tetracycline showed a significant prolonged anti-bacterial effect.Research results proved that the floating and releasing progress of core-shell beads can be well controlled by adjusting the shell layer formulation that could promote the function of gastroretentive drugs. Oral administration is the most common drug delivery method as it is multifunctional and convenient . The sizHelicobacter Pylori in vitro, but are not very effective for the eradication of infection caused by Helicobacter Pylori in vivo. The short residence time is the key problem [Helicobacter Pylori in stomach [The floating drug delivery system (FDDS) is also called a hydrodynamically balanced system (HBS). In FDDS, the drugs carriers float in gastric juice to ensure that drugs do not leave stomach shortly. It efficiently increases drug bioavailability by prolonging the release period. The variation of drug concentrations in blood is also decreased . FDDS is problem . Better  stomach , 6, whicCurrently, most FDDS would not float for 2–8 h; however, the drug float and release periods need to be prolonged. To obtain improved drug float and release times, previous research studies have widely investigated control of the drug carrier materials. Kawashima, Sato, Thanoo –10 et alIn this research, we prepared the core-shell floating particles for GDDS. We used two polymers, chitosan and xanthan gum, as the shell layer, which are cationic and anionic, respectively. Chitosan and xanthan gum were widely used in previous researches for the drug delivery, so their biocompatibility and biodegradability have been well approved. Manca et al. controlled the ratios of chitosan and xanthan gum to adjust the surface potential of liposome with coating layer, which influenced the rheological properties of microparticles in aerosol performance . In the Adjusting the chitosan/xanthan gum (C/X) ratio enabled us to adjust the polymeric entanglement, and allowed us to control the shell layer properties and structures in floating beads. We studied how the shell layer affects the float, release and biocompatibility properties. Moreover, the efficient drug carriers with better duration in GDDS would be developed by controlling the shell layer properties.Xanthomonas campestris), NaHCO3 (sodium bicarbonate powder), chitosan (low molecular weight), tetracycline (> 98.0%) were purchased from Sigma-Aldrich . CaCl2  and HCl and acetic acid were purchased from J.T. Baker, Japan. Dulbecco’s modified eagle medium (DMEM), fetal bovine serum (FBS), penicillin, trypsin was purchased from Gibco Life Technologies (Thermo Fisher Scientific - TW). Kaighn’s modification of Ham’s F-12 medium was purchased from Manassas, VA, USA. MTT -2,5-diphenyltetrazolium bromide) kit was purchased from Carlsbad, CA, USA. Distilled deionized (DI) water was used throughout the experiment.Alginic acid sodium salt (medium viscosity), xanthan gum  solution for shell. First, 50 mg tetracycline and 200 mg alginate were mixed in 10 ml DI water, and stirred for 2 h to obtain the tetracycline-alginate solution. 200 mg NaHCO3 was then added into alginate solution, and stirred at 500 rpm for 1 h.On the other hand, the C/X solution was prepared by mixing chitosan solution and xanthan gum solution. The chitosan solution was fabricated by dissolving chitosan powders in 1 v/v% acetic acid aqueous solution, and xanthan gum solution was prepared by dissolving xanthan gum in DI water. After that, chitosan and xanthan gum solutions were mixed with different C/X ratios of 4:1, 4:2, 4:3, 4:4 and 1:4, where CaCl2 was also added as the concentration of 1 w/v%.After finishing tetracycline-alginate solution for core and C/X solution for shell respectively, we extruded the tetracycline-alginate solution through a 26-gauge needle into the continuously stirred C/X solution. The C/X shell layer was then formed outside the alginate core, and the solution with floating beads was stabilized after continuously stirred for 15 min. After the beads were collected and washed with DI water, they were vacuum dried for 48 h and then stored in 4 °C until used. The exact formulations of core and shell layer were described in Table SEM  images were taken to analyze the morphology of floating beads. The beads were dried at 4 °C. After that, the bead samples were spread and fixed on the metal plate with double-sided carbon tape, and a gold layer was then coated on the sample surface under vacuum using an auto-sputter coater for 1 min under argon atmosphere. The beads morphologies were then observed with SEM. With SEM images, the bead diameters were measured by Image J software.The swelling percentage of beads was determined by measuring the extent of swelling of the polymer matrix in pH 2 aqueous solution. The weight of dried beads was recorded. After the immersion in pH 2 buffer for 1, 2, 4, 6, 8, 12 and 24 h, water on bead surfaces was removed by filter paper and the weight beads were measured again. The swelling percentage was calculated by using the following equation.f: number of floating beads; Ns: number of settled beadsFor the floating analysis, twenty dried beads were kept in pH 2 buffer for 24 h with continuous shaking. After 1, 2, 4, 6, 8, 12 and 24 h, the floating percentage was calculated according to following equation:The dry beads were immersed in pH 2 buffer in the dynamic gastric simulator . After 2In the analysis of encapsulation efficiency, beads were suspended in pH 2 buffer with continuous shaking at 37 °C for 24 h, where a part of encapsulated tetracycline was released. Then, the beads were completely broken by using ultrasonic shaker for 30 min, and the residual tetracycline would be completely released. After the solution was filtrated, UV/VIS spectrophotometer was applied to quantify tetracycline in buffer by deducting the absorbance at 266 nm, allowing the determination of encapsulation efficiency as following equation.3 cells per well. AGS has been applied in the cytotoxicity of biomaterials for stomach in previous researches [MTT assay was applied to evaluate the cytotoxicity of core-shell beads. In this experiment, a stomach cell line, AGS, was seeded on a 24-well plate with the density of 5 × 10searches . As the E. coli which was from Bioresources Conservation and Research Center (BCRC), Taiwan. The medium with cultured bacterial was added onto agar plate evenly, followed by an overnight culture. After that, core-shell beads were added onto the agar plate, and the antibacterial effects were observed at various time points.LB medium  was applied for the culture of The one-way ANOVA was used to analyze the statistical significance of particle diameters, encapsulation efficiency, floating percentage, and cell viability. The two-way ANOVA was used to analyze the statistical significance of swell percentage.2 and NaHCO3) and alginate in core were fixed.Table p > 0.1 for the formulation effects on particle diameters, revealing the ratios of chitosan and xanthan gum did not change particle size significantly. The cross-sectional images of core-shell beads are presented in Fig. The morphologies of core-shell particles were analyzed by using SEM as shown in Fig. p < 0.05 from ANOVA test) and reached a steady state over the next 2 h. The highest swelling ratios at steady state were 150–250%.Figure p < 0.05).Figure p > 0.15).Figure p < 0.1 and p < 0.05 from ANOVA test) at 2nd, 4th and 6th hr. That is, the particles released tetracycline faster when the amounts of chitosan are much higher or much lower than the amounts of xanthan gum, such as C/X = 4:1 and 1:4.The encapsulation efficiency and releasing rate of tetracycline of floating beads are described in Fig. The biocompatibility of floating particles was analyzed by culturing AGS cell line. The C/X = 4:3 core-shell beads were applied because they showed the best floating and long-term release properties in this research. The results in Fig. E. coli. It was caused by the released tetracycline, and the core-shell beads without tetracycline did not result in any anti-bacterial ring  were prepared by blending chitosan and polyanions, such as carrageenan [The densities of chitosan and xanthan gum used in this research were approximately 0.3 and 1.5 g/cmrageenan , alginatrageenan , poly , the release of tetracycline is also higher than those from C/X = 4:3 and 4:4. This is due to the strong interactions between positive chitosan and negative xanthan gum, which results in a dense shell layer. With the dense shell layer, the particle swelling is slow, and the tetracycline release is delayed due to the high resistance in mass transfer. The results supported that the prolonged release can be achieved by controlling the formulation of the dense layer in the core-shell floating beads.Many researches supported that the ionic crosslinking, which was achieved by mixing chitosan and polyanions, was effective on the control release –25. In tp < 0.005 from ANOVA test) and drug release (p < 0.1 and p < 0.05 from ANOVA test). This supports that the core-shell beads developed in this research can continuously delivery antibiotics for a long period for a certain period.The floating beads are proved to be biocompatible, and can carry effective antibiotics when they were applied. The released tetracycline from core-shell beads present clear antibacterial effects. According to the statistical analysis, the C/X ratio significantly prolonged the swelling (In this research, we developed core-shell floating beads for GDDS with porous alginate core and a dense chitosan/xanthan-gum shell layer. The compactness of the shell layer in floating beads was controlled by adjusting the ratios of anionic xanthan gum and cationic chitosan. When the C/X ratio was 4:3 and 4:4, the shell layer would be dense and would cause high resistance of mass transfer under the water penetration. Thus, a low swelling rate and a prolonged release was achieved. The experimental results proved the high biocompatibility of the floating beads, and the anti-bacterial effects of beads were also significant after the release for 4 h. This study proposed the method to modify the properties of shell layer, allowing the control of the swelling and release behaviors of floating beads.


Moreover, significant correlations were observed between the changes in DF ROM and passive torque at DF ROM in both ROLL and NO-ROLL. Changes in ROM after RM appear to be the result of sensory changes , affecting both rolled and non-rolled body regions. Thus, therapists and exercise professionals may consider applying remote treatments if local loading is contraindicated.In sports and clinical settings, roller massage (RM) interventions are used to acutely increase range of motion (ROM); however, the underlying mechanisms are unclear. Apart from changes in soft tissue properties , neurophysiological alterations such as decreased spinal excitability have been described. However, to date, no study has investigated both jointly. The purpose of this trial was to examine RM’s effects on neurophysiological markers and passive tissue properties of the plantar flexors in the treated (ROLL) and non-treated (NO-ROLL) leg. Fifteen healthy individuals  performed three unilateral 60-s bouts of calf RM. This procedure was repeated four times on separate days to allow independent assessments of the following outcomes without reciprocal interactions: dorsiflexion ROM, passive torque during passive dorsiflexion, shear elastic modulus of the medial gastrocnemius muscle, and spinal excitability. Following RM, dorsiflexion ROM increased in both ROLL (+19.7%) and NO-ROLL (+13.9%). Similarly, also passive torque at dorsiflexion ROM increased in ROLL (+15.0%) and NO-ROLL (+15.2%). However, there were no significant changes in shear elastic modulus and spinal excitability ( Improving flexibility is a frequent goal in sports. Besides representing a potential hallmark of athletic performance, it may also be related to injury risk. Previous studies reported an association of low flexibility , high muPrevious studies  have notConsidering the mixed evidence regarding local foam rolling effects, recent research has increasingly focused on neurophysiological changes. For instance, a previous study investigated the effect of rolling intervention on spinal excitability  and cortSummarizing the evidence regarding foam rolling mechanisms in a topical review, Against this background, this study was geared to (1) investigate the effects of roller massage (RM) on passive tissue properties and neurological changes as well as their relationship with ROM in the treated leg and (2) investigate the possible cross-education effect of the passive properties and neurological parameters.A randomized, repeated-measures experimental design was used to investigate the neural and tissue-specific effects of RM. The participants, in random order, visited the laboratory five times . The firThe protocol for the four measurement days was identical and based on a previous study . After tFifteen healthy adults volunteered to participate in this crossover study  Individuals with a history of neuromuscular disease and musculoskeletal injury involving the lower extremities were excluded. They were fully informed about the procedures and purpose of the study and provided written informed consent. The Ethics Committee of the Niigata University of Health and Welfare, Niigata, Japan (Procedure #18304), approved the study, which complied with the Declaration of Helsinki.∗Power  was used to perform an a priori sample-size calculation (primary outcome: ROM) for a repeated-measure analysis of variance (ANOVA)  based on a previous study . Participants were in a prone position on a treatment bed. An examiner moved the roller massager forward and backward, rolling on the muscle belly of the gastrocnemius. Speed was controlled with a metronome set to 60 bpm . At each beep, the examiner foam moved the massager one stroke up or down. The intensity was steered based on the participants’ feedback, targeting a 7/10 in discomfort on a numerical rating scale  . Based oThe participants were in the prone position with a 0° knee angle , having the foot of the measured leg attached to the footplate of a dynamometer. Using the passive mode, the device moved the ankle into passive dorsiflexion at a speed of 5°/s, starting from 30° plantar flexion  . Measure2 near the center of MG, with a 5-mm-diameter-circle analysis area at the center of the region (Shear elastic modulus of MG was measured by means of an ultrasonic shear wave elastography (SWE) imaging device  with a linear probe  and using the SWE mode . SWE provides a two-dimensional map of the shear elastic modulus on the B-mode image of the target tissue at 1 Hz with a spatial resolution of 1 × 1 mm . Measuree region . Elastoge region , the sheFor PPT measurements, we employed an algometer (NEUTONE TAM-22 (BT10); TRY ALL, Chiba, Japan). Participants adopted a prone position on the treatment bed similar to that for the DF ROM measurements. The joint position was 20° at the plantar flexion position. With continuously increasing pressure, the metal rod of the algometer was used to compress the soft tissue in the measurement area. Participants were instructed to immediately press a trigger when pain was experienced rather than just pressure. The value read from the device at this time point (kilograms per square centimeter) corresponded to the PPT. Based on previous studies , the meaElectromyographic activity of the soleus muscle was recorded using self-adhesive electrodes with recording surface electrodes . Before measurements, the skin was cleaned with alcohol to improve conductivity. Electrodes were placed on the soleus according to SENIAN , with thIn the spinal cord mononeuron excitability assessment, participants were instructed to lie in a prone position on the treatment bed with the knee at a 0° knee angle and the ankle in a 20° plantar flexion position. The participants were requested to relax completely, which we ensured by controlling EMG activity. In this study, electrical stimulations were delivered using a constant current stimulator (Isolator SS-104 J: Nihon Kohden Corporation) targeting the tibial nerve using 1-ms pulses. Based on previous researches , 2020, tmax) and M wave (Mmax) maximum amplitudes. Stimulations were increased by 0.5 mA every 10 s until Mmax was reached, with the corresponding intensity for all stimulations retained for analysis. The three stable Hmax and Mmax are measured, and the average value of each is used for analysis. The H/M ratio, an index of spinal excitability, was calculated from the measured Hmax and Mmax  was determined by the coefficient variation (CV) and intraclass correlation coefficient (ICC) using eight healthy students , with a 1-week washout between the two measures. The CV of and passive torque at DF ROM, PPT, shear elastic modulus at 0° and 20°, and H/M ratio were 9.9 ± 13.3, 6.4 ± 3.9, 4.0 ± 3.3, 2.0 ± 1.4, 3.6 ± 2.4, and 9.7 ± 8.7%, respectively, and the ICCs for the measurements were 0.96, 0.96, 0.97, 0.92, 0.91, 0.88, and 0.96, respectively.t-test. If a significant difference was noted between PRE values, we used a paired t-test with Bonferroni corrections to compare the significant differences between the PRE and POST values of each leg, and the Wilcoxon signed-rank test was used to compare the pre-post changes (in percentage) between the ROLL and NO-ROLL leg. Conversely, if no significant difference was noted between PRE values, a two-way repeated-measure ANOVA (time [PRE vs. POST] × legs [ROLL leg vs. NO-ROLL leg]) was used to investigate the interactions and main effects in the experiment. If a main effect for time (without interaction) was noted, significant differences between the PRE and POST values were determined using a paired t-test. The effect size was calculated as a difference in the mean value between PRE and POST divided by the pooled standard deviation (SD)   with efft-tests showed a significant difference in PPT between the PRE values of the ROLL and NO-ROLL legs  but not for dynamic stiffness, shear elastic modulus, and H/M ratio (p > 0.05). According to post hoc testing with paired t-tests, DF ROM and PPT increased after RM (p < 0.01). In addition, regarding PPT, paired t-tests with Bonferroni corrections showed a significant increase after RM intervention in both ROLL and NO-ROLL sides, but the Wilcoxon signed-rank test revealed no significant difference in the relative pre–post changes between the ROLL and NO-ROLL leg  in any other outcome , whereas no significant associations occurred between changes in DF ROM and PPT , dynamic stiffness , shear elastic modulus at neutral , 20° plantarflexion , and H/M ratio  at both legs.Significant correlations were observed between the changes in DF ROM and passive torque at DF ROM in both ROLL and NO-ROLL legs , myofascial restriction reduction, fluid changes, and cellular responses . No signFor the ROM change noted after the RM intervention, Our correlation analysis showed a significant association between the changes in DF ROM and the passive torque at DF ROM in both ROLL and NO-ROLL  but not Other factors involved in the ROM change could be changes in muscle stiffness and/or spinal excitability. However, the current study showed that RM intervention increased ROM, with no changes in shear elastic modulus and H/M ratio, i.e., mechanical property and neurological change, suggesting that muscle stiffness and H/M ratio changes were not related to DF ROM increase in both legs after the RM intervention. This study had several limitations. In the previous study , H-refleThis study investigated the effect of RM on DF ROM, passive torque, shear elastic modulus, PPT, and H/M ratio on both the intervention and non-intervention sides. The results showed that DF ROM and PPT increased after treatment in both legs. The increases in ROM were related to the changes in stretch tolerance but not muscle stiffness, PPT, and spinal excitability. Interestingly, this study showed the same degree of RM intervention effect on DF ROM and the intervention and non-intervention sides. These results could be a treatment option in clinical and sports settings. Specifically, unilateral RM intervention on the healthy side may maintain or increase the ROM of the disabled side even if RM intervention could not be adaptable owing to injury or pain.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by this study was approved by the Ethics Committee of the Niigata University of Health and Welfare, Niigata, Japan (Procedure #18304), and has complied with the Declaration of Helsinki requirements. The patients/participants provided their written informed consent to participate in this study.MN contributed to the study design and data collection, and drafted and critical revision to the manuscript. RK, SS, KaY, RY, KoY, YM, and FS contributed to the data collection and made critical revisions to the manuscript. AK and JW contributed to the study design and data analysis and made critical revisions to the manuscript. All the authors approved the final version of the manuscript and agreed to be accountable for all aspects of the work.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.