The number of people living with HIV (PLHIV) in need of treatment monitoring in low-and-middle-income countries has been rapidly expanding, placing an increasing burden on laboratories. Promising new point-of-care (POC) test have the potential to reduce laboratory workloads, but the implementation cost is uncertain. We sought to estimate the costs of decentralized POC testing compared to centralized laboratory testing for PLHIV initiating treatment in South Africa.We conducted a microcosting analyses comparing clinic-based POC testing to centralized laboratory testing for HIV viral load, creatinine, and CD4 count monitoring. We completed time-and-motion studies to assess staff time for sample collection and processing. Instrument costs were estimated assuming five-year lifespans and we applied a 3% annual discount rate. Total costs and cost per patient were estimated over a five-year period: the first year of ART initiation and four years of routine HIV monitoring, following World Health Organization ART monitoring guidelines.We estimated that per-patient costs of POC HIV viral load, CD4, and creatinine tests were USD $25, $11, and $9, respectively, assuming a clinic volume of 50 patients initiated per month. At centralized laboratories, per-patient costs of POC HIV viral load, CD4, and creatinine tests were USD $26, $6, $3. Total monitoring costs of all testing over a 5-year period was $45 higher for POC testing compared to centralized laboratory testing ($210 vs $166).POC testing for HIV care and treatment can be feasibly implemented within clinics in South Africa, particularly those with larger patient volumes. POC HIV viral load costs are similar to lab-based testing while CD4 count and creatinine testing are more costly as POC tests. Our cost estimates are useful to policymakers in planning resource allocation and can inform cost-effectiveness analyses of POC testing. Nearly 37 million people living with HIV (PLHIV) worldwide are either receiving or are eligible to initiate antiretroviral therapy (ART). The WorlPoint-of-care (POC) testing can enable HIV monitoring to be conducted onsite at local clinics, thereby relieving the burden on laboratory facilities, providing rapid delivery of results to patients and providers, and improving linkage to care.–9 FurtheSouth Africa has the largest HIV epidemic in the world, with an estimated 7.9 million PLHIV ,18. The We conducted a detailed microcosting analysis from the clinic perspective to compare the costs of HIV monitoring using decentralized POC tests to the costs of performing equivalent tests at centralized laboratories. Our costing analysis was conducted within the Simplifying HIV TREAtment and Monitoring (STREAM) study, a randomized clinical trial of POC HIV VL monitoring in Durban, South Africa. In STREAEthical approval for this study was granted by the University of Washington Institutional Review Board (STUDY00001466) and the University of KwaZulu-Natal Biomedical Research Ethics Committee (BFC296/16).We conducted a microcosting analysis to identify materials and instruments used to conduct POC tests and collect samples for both POC and centralized laboratory tests. Materials costs relevant to both POC and laboratory tests included consumables required to collect samples at the clinic and staff salary spent on obtaining blood samples. Blood samples for POC creatinine tests were obtained through capillary finger pricks, while samples for POC CD4 count and HIV VL tests and all centralized laboratory tests were obtained through venous blood draws.Consumables common to all samples included alcohol swabs, gloves, and adhesive bandages. Samples obtained through blood draws additionally required syringes, needles, and blood tubes. Consumables specific to POC creatinine tests included a lancet and a test strip. Consumables specific to POC HIV VL and CD4 count tests included test cartridges.™ CD4 ; and StatSensor® Xpress-iTM Creatinine POC .This study utilized the following POC testing instruments: Xpert® HIV-1 Viral Load IV ; PimaPoint-of-care instrument/maintenance costs included: POC test instruments; computers and software; quality control (QC) reagents; refrigerator; and value of laboratory space. We estimated instrument and maintenance costs assuming an instrument lifetime of 5 years, as is common for costing analyses based on WHO guidelines and other published costing literature. FollowinCosting data for centralized laboratory tests were obtained from the South African National Health Laboratory Service (NHLS) 2017 price lists. The cost charged to a clinic per test was inclusive of sample transport, laboratory staffing costs, laboratory consumables, instruments, and maintenance. Therefore, for study purposes, centralized laboratory test costs were estimated from the cost charged to clinic per test, plus the cost of professional nurse time and cost of clinic consumables required to collect a sample.We recorded clinic staff time costs as staff salary per minute multiplied by time spent directly involved with sample collection and/or processing. The following activities were observed:Clinical nursing staff time needed to collect samples (time required to draw blood and complete paperwork).POC test laboratory technician time needed to perform POC HIV VL and CD4 count tests in clinic-based POC test laboratory facility;Clinical nursing staff time needed to perform POC creatinine testsTotal time for POC tests to be conducted on their respective instruments, andTime to conduct quality control checks.We observed 20 patient blood draws to assess time required to obtain blood samples and complete paperwork, and completed 10 individual observations of staff conducting each of the POC creatinine test and the POC HIV VL test . We computed staff time costs for POC testing by observing the total time needed and removing non-active waiting time during POC test processing as the staff completed routine clinic activities during this time. As staff time required to run tests differed by only a few seconds during the observation period, we averaged observed times to estimate staff time instead of using the range as this would negligibly change the per test cost.To estimate the total cost of initiating and monitoring a patient on ART for 5 years, we followed the WHO testing and monitoring guidelines to establish the number and types of monitoring tests conducted for one PLHIV over a 5-year period. These guidelines are utilized by the South African Ministry of Health for initiating and monitoring PLHIV on ART :23]23]ART Initiation Visit: CD4 Count, CreatinineMonth 3: CreatinineMonth 6: Creatinine, HIV VLMonth 12: CD4 Count, Creatinine, HIV VLAnnual Visit After First Year: Creatinine, HIV VLWe assumed the following: the clinic has a stable power supply; POC test material and instrument prices reflect preferential pricing provided by manufacturers for LMICs; patients attended all monitoring visits recommended by WHO guidelines. We did not include utility costs in the POC test costs. Finally, based on consultation with in-country staff, we assumed a typical large South African clinic initiates an average of 50 patients onto treatment per month. However, we varied this assumption in sensitivity analyses. We also conducted a sensitivity analysis excluding creatinine costs since the clinical value of creatinine monitoring is under debate. Further, changes in ART treatment regimens are replacing medications that need creatinine monitoring with new drugs that don’t require routine creatinine monitoring. Finally, we conducted a sensitivity analysis that assumed an instrument lifespan of 10 years.Staff salary figures and costs of materials and instruments were obtained from published government records, laboratory price lists, clinic invoices, and health economic literature . We calcStaff costs including overhead supervision, external quality control checks were not included in our costing estimates. Since the data was collected at a research clinic with a fully functional in-clinic laboratory, it was not possible to estimate the time supervisors spend overseeing tests and how much supervision time would be required in a traditional clinical setting.Individual per-test costs for POC CD4 count, HIV VL, and creatinine tests were estimated to be $11, $25, and $9 respectively, when performed as POC tests at a clinic initiating 50 patients per month for 5 years . The larThe combined cost of instruments, clinic space, and annual maintenance for decentralized POC HIV VL, CD4 count, and creatinine testing over a 5-year period was estimated to be $36,414. After discounting costs pf capital costs, the total cost of increases to $72,084 over a 5-year period. The largest contribution to the cost was the GeneXpert® instrument, at $21,762 . Total uAs the equipment component of the total test cost per depends on how many tests are conducted, we calculated the cost per patient at varying patient volumes. The cost of initiating and monitoring one PLHIV on ART for a total of 5 years was estimated to be $349 for a clinic initiating 10 patients per month, $211 for a clinic initiating 50 patients per month, and $193 for a clinic initiating 100 patients per month compared to $166 for centralized laboratory testing at any clinic size . PatientAssuming creatinine testing is removed from the testing schedule, the cost of initiating and monitoring one PLHIV on ART for 5 years using POC tests is estimated to be $281 for a clinic initiating 10 patients per month, $149 for a clinic initiating 50 patients per month, and $132 for a clinic initiating 100 patients per month .We also conducted a sensitivity analysis assuming a longer instrument lifespan of 10 instead of 5 years. Under this scenario, the cost of initiating and monitoring one PLHIV on ART for a total of 10 years was estimated to be $414 for a clinic initiating 10 patients per month and $339 for a clinic initiating 50 patients per month . With a For a 5-year instrument lifespan, instruments costs accounted for 50% of total cost for a clinic initiating 10 patients per month and 16% of total cost for a clinic initiating 50 patients per month .Per-patient costs for CD4 count, HIV VL, and creatinine tests were found to be $6, $26, and $3, respectively, when performed at a centralized laboratory . The totIn this microcosting study, we estimated the annual cost of implementing POC tests for ART monitoring to be approximately $45 higher per patient compared to using centralized laboratories over a 5-year period. The per-test cost of implementing POC VL testing was approximately the same as centralized laboratories while POC CD4 count and POC creatinine testing $5 and $5 higher per test respectively, compared to laboratory testing. These cost estimates assume a clinic initiates 50 patients per month, which is typical for moderate-size clinics in South Africa. At lower clinic volumes, POC testing became more expensive; for a clinic initiating 10 patients per month, POC testing would be about $183 more expensive per patient over 5 years. These costs include a 3% discount on all instruments and lab space. Discounting costs were not included in centralized laboratory costs as these individual costs were not evaluated in our study. Therefore, it is likely that total equipment costs in this study are a conservative estimate. Overall, we find that POC HIV VL testing can be feasibly implemented into routine care at reasonable costs with minimal processing time for healthcare staff.Per-patient POC testing costs were highly sensitive to the number of tests performed on each instrument. This is affected by clinic volume and lifespan of instruments. While HIV VL POC costs were comparable to centralized laboratory costs when implemented at moderate to high volume facilities, they were substantially higher than centralized laboratory costs at lower volumes, (e.g. 10 patients initiated on ART per month). Therefore, facility size will likely be an important consideration as countries decide how to scale up POC technology. Costs were also much lower when an instrument lifespan of 10-years and were comparable to centralized laboratory testing.Creatinine testing was a significant contributor to total POC testing costs, as it is more than twice as expensive to perform as a POC test compared to a centralized laboratory. However, recent evidence suggests that testing creatinine levels in asymptomatic patients on treatment regimens such as tenofovir disoproxil fumarate has limited patient care benefits in low-resource settings. FurthermOur cost estimates are similar to those of previous studies.,28 FactoAn important consideration impacting POC testing costs in South Africa is the potential for using GeneXpert® instruments in clinics to test for TB and sexually transmitted infections, in addition to HIV VL POC testing. Investment in the GeneXpert® instrument would be shared among more patients, improving the potential cost savings of integrating POC testing into clinics and increasing health benefits to patients from TB screening. Since the GeneXpert® instrument is the largest single instrument cost considered in this analysis, this can significantly reduce POC testing cost particularly in clinics with smaller patient volumes which may not be using the instruments at capacity and who would benefit most from increasing the volume of tests performed on instruments.Additionally, the GeneXpert® instrument is available in varying sizes ; this flexibility can allow clinics to scale the instrument size and corresponding costs to their needs, potentially reducing the cost per POC test. Finally, the costs of instruments for this study were based on prices quoted to a research facility. Manufacturers often offer reduced costs for LMIC and public institutions. For example, a WHO report on the implementation of the GeneXpert® instrument for TB testing estimated the installation costs of the GeneXpert® instrument, combined with annual maintenance costs, to be about $21,700 over 5 years, approximately $2,400 lower than the 5-year costs estimated in this study. Another There are several strengths and limitations to this study. First, this study does not include the costs of scaling up implementation to a regional or national scale. For example, administrative costs to running POC programs may not contribute significantly to per test costs, they will add significant costs when considering large-scale implementation. Further research will be required to estimate these additional costs. Second, this study was carried out in a single research facility which employed dedicated laboratory and staff to perform POC HIV VL and CD4 count tests, and our results may not be representative of clinics without specialized staff and facilities. In settings where staff have less training, time needed to conduct laboratory testing may be longer, environmental conditions in some clinics may be less ideal for these instruments causing more wear and tear which can shorten their life-span. Finally, we may not have captured all relevant costs: for example, the costs of POC test cartridge disposal , and a combination of the two. Using these different formats for POC testing delivery, tailored to the needs of the region, they concluded that POC testing was able to reduce the costs of HIV monitoring testing by 6–35%. This was primarily accomplished by reduction in transportation costs and better instrument utilization. Our study similarly found that POC testing per test costs were highly variable based on number of tests performed on an instrument. Removing inefficiencies and equipment sharing between clinics appears to help decrease costs in smaller, more remote clinics unable to absorb the costs of equipment individually. Finally, while we did not evaluate costs related to increasing distance from centralized laboratories, this will likely be an important cost factor in rural clinics as recent research has shown that transportation costs represent up to 34% of the costs of HIV VL monitoring testing transported to centralized laboratories. [Our study shows that POC technology has the potential to be a financially viable method for clinics with moderate to large patient loads. However, At smaller clinic loads, the upfront costs of instruments and their maintenance makes POC testing more expensive compared to centralized laboratory testing. Further research is needed to evaluate POC testing in non-urban settings, as well as non-research settings. Future microcosting studies of centralized laboratory costs can provide a more detailed understanding of unit cost components compared to our analysis which utilized published costs estimates. In addition, studies are needed to project the cost-effectiveness of POC testing across various clinic volumes. Point-of-care testing offers an opportunity both to improve patient access to testing and monitoring services at reasonable costs.S1 File(PDF)Click here for additional data file.S2 File(PDF)Click here for additional data file.S3 File(PDF)Click here for additional data file.S1 TableAn outline of the assumptions and data sources used for each category of costs.(PDF)Click here for additional data file.S2 TableA summary of per test and per patient costs as described in the manuscript but without discount costs added.(PDF)Click here for additional data file.


The authors report a 1 month follow up of a neonate described as “miracle baby” because she was born Ebola virus disease-free and survived after her mother was infected with Ebola virus during the third trimester of pregnancy.This female newborn baby was registered at the Maternity of Beni Reference General Hospital and the Ebola Treatment Centre in eastern Democratic Republic of Congo. She was delivered normally and showed no signs of Ebola infection. All tests were negative for Ebola. At 1 month follow up, the baby is growing normally.This very rare happy outcome for neonates of mothers infected with Ebola virus motivated the authors to report the case. According to the World Health Organization (WHO), Ebola virus disease (EVD) is a serious, highly contagious and fatal disease in humans. The virus is transmitted to humans from wild animals and spreads to the population through interpersonal transmission .Ebola virus disease during pregnancy is associated with a very high rate of obstetric complications and perinatal adverse outcomes including spontaneous abortion, premature rupture of membranes, premature delivery, ante- and postpartum haemorrhage, intrauterine death of the foetus, maternal and neonatal death . NeonateThe data show that intrauterine content remains positive for Ebola virus ribonucleic acid (RNA) . Since tDuring the 2014–2016 outbreak, Médecins Sans Frontières treated at least 54 EBOV-infected women with pregnancies in the second and third trimester and recorded 35 second-trimester miscarriages and deliveries in Ebola treatment centres. The single baby born alive died 2 days after birth. To date, there is only one survival beyond neonatal period reported in the literature about babies born to EBOV-infected women. It was a female baby born with congenital EVD who was followed up for 12 months of life in Conakry (Guinea) and was found to be growing normally . While iHere we report a 1 month follow up of a neonate born to a mother who survived the Ebola virus disease in eastern Democratic Republic of Congo during the Ebola outbreak that was declared in August 2018 and continues to wreak havoc in the northern areas of the province of Nord-Kivu and the neighbouring areas in the province of Ituri.Our patient’s mother, 22-year-old previously healthy multigravida woman  was admitted to the Beni General Hospital Ebola Treatment Center 2 days after onset of symptoms  and was confirmed positive for EBOV by polymerase chain reaction (PCR). She reported a 34 week pregnancy and confirmed that the previous pregnancy follow up was uneventful. She was discharged after 1 week of management made of rehydration, Cefixime, Paracétamol and other supportive measures.At the gestational age of 38 weeks, vaginal delivery occurred after a 10 h labor, the foetus was in breech presentation with meconial amniotic fluid and a normal placenta. The female newborn baby did well initially with an Apgar score of 8/9/10. General and neurologic examination did not reveal any pathology. The vital signs and anthropometric parameters at birth were as follows: temperature 36.5 °C, regular breathing at 52 cycles per minute and heart rate 146 beats per minute; weight of 3500 g, head circumference of 36 cm and height of 53 cm. The haemoglobin level was 16 g / dl; the umbilical cord blood PCR, the blood and salivary swabs were negative for the Ebola virus disease. She was subjected to an antibiotic therapy made of cefotaxime (3x200mg/day) and was discharged after 5 days.At 1 month and 6 days of age, the baby was growing normally, she weighed 4100 g , she was being breastfed and her mother reported no illness in the past days of her life.We describe a neonate who was born EBOV-free from a mother who survived Ebola virus disease during her pregnancy in late stage. We think that this favourable outcome was probably due to the fact that the mother presented a relatively mild disease phenotype which was not transmitted to the foetus in utero.Although information on the consequences of Ebola during pregnancy is limited (in terms of risk factors), early reports seemed to indicate very high mortality, which also leads to a fatalistic approach to the pregnant population management , 9. HoweAccording to very disturbing reports from West Africa, pregnant women suspected of having Ebola virus were denied access to health facilities because healthcare providers feared they would infect other patients. Because of the history of an almost uniformly fatal outcome in infants born to mothers with Ebola, these pregnant mothers and their infants have not been given the care needed . HoweverAlthough these are very rare cases, some pregnant women have survived the Ebola virus disease without losing the unborn child. In a medical facility in developed countries, the chances of survival of a pregnant woman with Ebola virus disease could be high thanks to the implementation of standard supportive care. However, whether supportive care for neonates born to women infected with EVD would be effective in high income settings also remains to be seen .Ebola virus disease remains a serious public health problem because of its high mortality and the fatal obstetrical and neonatal complications. Our case highlights the possibility of delivery and survival of disease-free full term babies born to mothers who survived the Ebola virus disease. Thus, care should be strengthened in pregnant women to improve the chances of saving neonates born in these conditions.


Potentilla fulgens root along with RAN. Since evaluation of anticancer role, if any, of P. fulgens on RAN-mediated genetic alterations in human is difficult because of other compounding factors, this study was undertaken in mice to focus on gastric carcinogenesis since ad libitum administration of RAN extract with lime in drinking water induced stomach cancer due to greater exposure of its lining. A total of 160 mice were used at different time points and either methanol extract of P. fulgens roots (PRE) or mixture of four compounds of ethyl-acetate fraction (EA-mixture) was mixed with mice feed. Histological studies revealed that RAN + lime induced cancer in all the mice and interestingly only 20% developed cancer when PRE/EA-mixture was provided along with RAN + lime. Higher frequency of precocious anaphase and over expression of p53 and Securin genes were significantly reduced by PRE/EA-mixture. Thus PRE/EA-mixture mitigates the RAN-induced tumor-initiating process in stomach by maintaining expression of tumor suppressor and check-point genes under control.People of north-eastern states of India consume raw areca-nut (RAN) and lime which could lead to oral, esophageal and gastric cancers. However, the incidence of these cancers are significantly lesser in those who consume pieces of  Traditionally, people of these regions consume betel-quid which contains raw areca-nut (RAN), lime and a small part of betel-leaf but without tobacco. The whole betel-quid is swallowed after chewing and such a habit has been associated with development of these cancers4. A high frequency of gastric cancer in this region has also been associated with, besides RAN and lime, several causative factors like some non-vegeterian food items and alcohol. It has also been reported that Helicobactor pylori infection enhances development of gastric cancers caused by chewing and swallowing RAN and lime5. In animal model also, tumours have been shown to be induced in esophagous, stomach and liver when RAN extract was administered either by oral intubation6 or mixed with the diet7 or by ad libitum admisteration of RAN extract with lime in drinking water8. In earlier studies, induction of precocious anaphase (premature separation of sister-chromatids) and higher expression of p53 and Securin in non-target tissues like mouse bone marrow cells (BMC) were observed to be significantly associated with an increased risk of RAN-induced gastric cancer8. Subsequently, similar observation was made in human peripheral blood-lymphocytes of both oral and esophageal cancer patients and non-cancerous persons having the habit of consuming RAN9.Highest incidence of oral, esophageal and gastric cancers has been reported in the North-Eastern states of IndiaPotentilla fulgens roots along with the betel-quid show an extremely low incidence of these cancers11. Potentilla fulgens Wall. Ex Hook, commonly known as Himalayan Cinquefoil, is an important medicinal plant, because of its high ethnomedicinal value is well domesticated with the roots being commercially sold12. Potentilla, a member of Rosaceae family with about 500 species distributed across the world is nontoxic to human15. In Khasi Hills of Meghalaya, India. P. fulgens root-stock and whole herb is utilized as astringent and tonic for curing gum and tooth ailments, diarrhoea, diabetes and several other ailments16. The root extracts of P. anserine is used as a medicinal herbs against certain viral infection in Tibet17. P. anserine and other species have been used in traditional medicine in different parts of Europe and Northern America18. In China, different species of Potentilla have been used as folk medicinal herbs to treat diarrhoea, hepatitis and scabies19.It is significant to note that some people in these regions who consume pieces of 20. Significant anticancer activity has been reported for a number of triterpenoids compounds isolated from the aerial parts of Potentilla chinensis and the roots of Potentilla multicaulis21.Natural products have been widely studied because their potential roles in prevention and treatment of cancer. By monitoring several well-studied markers and cellular processes for colorectal cancer it was demonstrated that natural product effectively mitigates the cancer developmentP. fulgens is significantly low, it is of interest to establish a causal mechanism. Since it is difficult to evaluate its anticancer role in RAN-mediated cancer in human system because of interference by other compounding factors like tobacco, alcohol consumption and diverse food-habits, many studies on anticancer effects of different Potentilla sp have been examined in human cancer cell lines23. The present study, we have examined the potential anticancer role of P. fulgens root extract (PRE) and its ethyl-acetate (EA) fraction on RAN associated gastric carcinogenesis in mice in vivo. Our present results confirm that PRE or four compounds of EA-fraction indeed have anticancer potentialities since their co-administration delays the RAN and lime induced gastric carcinogenesis.In view of the above noted observation that the incidence of cancer in persons exposed to RAN and lime together with roots of In total, 160 mice were used at different time points for different experimental analysis Table , hyperplContinuous exposure to RAN extract with lime induced chromosomal instability which was evaluated in mouse BMC. We scored metaphase spreads 3 h after colchicine treatment and the data revealed a gradual but significant increase in the frequency of prematurely separated sister chromatids Fig.  in RAN +An ELISA method was used to detect 8-OHdG as a marker of oxidative DNA damage in DNA extracted from cells that were collected from the inner layer of esophagous and stomach from untreated control, RAN + lime and PRE + RAN + lime treated mouse for 180 and 260 days (5 mice per point) Fig. . It was We examined the expression of Mad2 and AuKA genes in cells from the inner layer of stomach of mice untreated (n = 10) or administered RAN + lime with or without PRE for 180 and 260 days (n = 6 per point). The quantitative RT-PCR results Fig.  showed tLevels of p53 and Securin in cells from inner layer of esophagous and stomach of those mice which were untreated (n = 5) or administered RAN + lime with or without PRE for 180 and 260 days (n = 5 per point) were evaluated by immunoblotting. Similar analysis was also performed in mice administered RAN + lime with or without ECGU for 180 days. Results indicate that the level of these proteins were elevated significantly in both the tissues after administration of RAN + lime. However, such elevated level was reduced significantly in presence of either PRE or ECGU along with RAN + lime Fig. .Figure 3We studied p53 and Securin expression by immunostaining in a panel of mouse stomach samples collected from untreated (n = 5) and treated with RAN + lime (n = 10) or PRE + RAN + lime (n = 10) for 260 days Fig. . The secP. fulgens root-stock and herb as folk-medicine against various ailments16. Evaluation of the total phenolic and flavonoid contents of P. fulgens root extract revealed the gallic acid and quercetin to be 138.8 ± 1.6 mg and 401.6 ± 4.6 mg per gm, respectively22. The root extract increased the survival of mice bearing Ehrlich ascites tumour cells and reduced cell viability in a dose-dependent manner in MCF-7 cells22. The ethyl-acetate (EA) soluble fraction of methanolic extract of P. fulgens root exhibited more efficient growth-inhibitory effect in MCF-7 and U87 cancer cell lines than hexane and n-butanol soluble fraction23. This observation prompted further purification of EA-fraction and a total of nine compounds including two new ursane type triterpenoids, Fulgic acid A and Fulgic acid B with a good antioxidant activity were identified24. Since the natural compounds act through diverse mechanisms and show minimal undesirable side effects25, it is attractive to evaluate anticancer activities of natural plant-based compounds in mammalian system.Natives of north-eastern regions of India use ad libitum in drinking water with dose increased every two months to mimic the human habit of consumption of RAN. In earlier studies with similar mode of treatment, parameters like precocious anaphase and expression of Securin and p53 genes were monitored at different time-points starting from 30 days to 300 days for understanding the process of RAN-induced carcinogenesis8. It was found that after 240 days of ad libitum administration of RAN extract with lime in drinking water all the mice developed gastric tumour. Therefore, in the present study we monitored the influence of PRE on carcinogenesis after 180 and 260 days of treatment.In this study, mice were given the whole RAN-extract and lime in vivo. These four compounds were selected primarily because of their higher yields: E (2.7%), C (1.6%), G (3.6%) and U (3.3%) in crude methanol extract24. Following the practice in Potentilla based traditional Chinese medicine26, we provided PRE or ECGU to experimental mice one month before RAN + lime treatment in order to strengthen their physiological system. In fact, it was suggested that such pretreatment of Potentilla based traditional Chinese medicine is fortifying the stomach, promoting the production of body fluid, invigorating the vital energy and nourishing the blood26.We examined the anticancer potency of PRE or of a mixture of epicatechin (E), catechin (C), gallic acid (G) and ursolic acid (U). in mice 27. Further, transition metal ions such as Cu−2+, Mn++, Fe2+ and Fe3+ present in RAN and betel-leaf, facilitate the production of more reactive oxygen species which in turn contributes to initiation and promotion of cancer28. DNA is biologically significant target of oxidative attack, and it is widely believed that continuous oxidative damage to DNA is a significant contributor to the development of the major disorders including cancers. Among the diverse oxidative DNA damages, 8-hydroxy-deoxyguanosine (8-OHdG) is one of the most important DNA lesions and has been used as a marker for DNA oxidation29. Present results indicate that the high level of 8-OHdG following RAN + lime treatment was considerably reduced when PRE was co-administered. Thus, the presence of PRE or ECGU reduced DNA damages induced by RAN + lime mediated reactive oxygen species because both EA and butanol fractions of PRE showed a significant antioxidant potential, as evaluated by DPPH• and ABTS + • (hydrogen atom transfer based) and MTT assay system24. Moreover, Corosolic acid which is present in EA-fraction of PRE, is known to deplete the level of 8-OHdG and reduce the oxidative stress30.Autoxidation of areca-nut polyphenols at alkaline pH and consequent generation of hydrogen peroxide and superoxide radicals accelerates RAN-induced carcinogenesis31. As observed earlier8, present results also showed a gradual increase in the frequency of precocious anaphase and aneuploidy in the BMC of mice following RAN + lime administration. Very significantly frequencies of both were reduced following co-administeration of both PRE or ECGU. It is known that the percentage of aneuploid cells correlates with the severity of premature sister-chromatid separation, suggesting a direct relationship between the severity of loss of checkpoint control and chromosome mis-segregation31. Partial loss of spindle assembly checkpoint gene like MAD2 in Hct116 cells and also in mouse primary embryonic fibroblast cells showed increased premature sister chromatid separation in the presence of spindle inhibitors and higher rate of chromosome mis-segregation events in the absence of spindle inhibitors31. Another study, showed that arecoline, an alkaloid component of areca-nut, upregulated the spindle assembly checkpoint genes like Aurora A, BubR1 and Mps1 which led to mitotic spindles distortion and misalignment of chromosomes32. In this study, downregulation of MAD2 and AuKA in stomach cells of the mouse was noted after administeration with RAN + lime but in the presence of PRE, their expression was similar to untreated control. Thus, the inhibitory effect of RAN + lime on some of the spindle assembly checkpoint genes can be abrogated significantly by either PRE or ECGU.Earlier studies highlighted the importance of precocious anaphase, which leads to aneuploidy, as a potential screening marker for identification of mitotic checkpoint defects during early days of RAN exposure in both mice and human34 and also in oral dysplastic lesions and therefore such alteration in p53 is considered to be an early event in oral carcinogenesis35. Securin or pituitary tumor transforming gene, is known to be involved in the regulation of chromatid separation at metaphase-anaphase interface of the cell cycle36. Overexpression of Securin has been associated with the aneuploidy formation due to chromatid mis-segregation and is demonstrated in multiple cancer types38. Overexpression of p53 as well as Securin genes in non-target cells like mouse BMC and in human peripheral blood lymphocytes as well as in stomach and esophageal cells after exposure to RAN + lime has also been reported30. Interestingly, similar elevated expression of both these genes in stomach as well as in esophageal cells seen in this study after RAN + lime treatment was found to be considerably reduced by PRE or ECGU exposure. Since cancer induction mainly happened in stomach due to the greater exposure, further immunohistochemical analysis was performed only in stomach, which fully agreed with the immunoblotting data.Elevated expression of p53 protein is well documented in head and neck squamous cell carcinomaP. fulgens root is consumed with RAN + lime. It is interesting that the mixture of four compounds of EA-fraction showed similar potentiality in anticancer property as of the PRE. Both catechin and epicatechin are well known natural antioxidant and showed anticancer property40. Gallic acid can induce cell death of various cancer cells41 by depleting cellular glutathione and blocking EGFR signal pathway42. A pentacyclic triterpene ursolic acid has also been shown to have anti-inflammatory, antioxidant, and antitumor effects43. Consequently, all these compounds show anticancer potentialities with different targets.In agreement with the above, only 20% mice developed RAN + lime induced stomach cancer in presence of PRE whereas in its absence all the RAN + Lime exposed mice developed cancer. Thus the presence of PRE or ECGU delays the RAN + lime induced stomach carcinogenesis by maintaining the normal expression status of tumor suppressor and check-point genes. This seems to explain the greatly reduced incidence of oral and esophageal cancers in human when in vivo study thus demonstrated that the presence of either PRE or ECGU mitigates the RAN + lime induced tumor-initiating processes. Most strategies on drug development use either combination of several monotargeted drugs or a compound with multitargeting properties20. A recent, multiscale model for association of frequently mutated genes and inflammation mediated stomach, colon and liver cancer, based on information retrieved from the Gene Expression Omnibus, the Cancer Genome Atlas, and Gene Ontology database has suggested two stages of inflammation-induced tumorigenesis44. The present study, in agreement with earlier results23 indicated that PRE or ECGU targets precancerous state as well as the transition from the precancerous to tumorigenic states. Further in-depth studies are needed to identify the driver pathways targeted by PRE or ECGU in inflammation-induced tumorigenesis.This Potentilla fulgens were collected from ten different plants from Shillong peak regions of Meghalaya state of India  after obtaining a proper approval of the forest officer. Taxonomic identification of the plant material was kindly confirmed by senior taxonomist of Department of Botany, North-Eastern Hill University, Shillong and a voucher specimen was deposited in the herbarium of the department (accession number 11906).Roots of 45. Chemical structures were elucidated by spectroscopic methods, especially ESIHRMS and 2D NMR techniques45. In brief, P. fulgens roots (1 Kg) were shade dried and extracted with methanol using Soxhlet apparatus and obtained 250 gm of crude Potentilla root extract (PRE) which was suspended in water-methanol mixture (80:20) and partitioned with hexane, chloroform, ethyl acetate (EA), butanol and water soluble fractions. The EA-fraction was further subjected to vacuum liquid chromatography in silica-gel using hexane-ethyl acetate (0 to 90% EtOAc) and then chloroform-methanol gradients (1% to 50% MeOH) as elutent to yield five major fractions. It was already reported that nine compounds including new triterpenes and phenolics were identified and characterized from this EA-fraction of the methanolic root extract of P. fulgens13  and controlled lighting . Standard mouse diet  and water ad libitum were used in all experiments. The experiments were conducted in compliance with institutional guidelines and approved by North-Eastern Hill University “Institutional Ethics Committee ” Board.ad libitum in the drinking water with lime (pH 9.8)  were used instead of PRE. A stock solution (5 mg/ml) of each compound was made and from this 10 µl of each was added to 6 gm of mice feed and finally made the tablet for each mouse. This way each mouse was consumed 50 µg of each compound every day.For preparation of metaphase chromosomes, bone marrow cells (BMC) were collected 3 h after colchicine 15 mg/kg) treatment from 5 untreated mice as control. BMC were collected from untreated and 60, 120 and 180 days of different treated groups. The number of mice were used in each treated group are shown in Table 5 mg/kg tAround 100 well spread metaphase plates were studied for each mouse. We performed chromosome counts on metaphase spread. Values are expressed as mean ± SEMs.™ Oxidative DNA damage ELISA kit, Cell Biolabs Inc.  according to manufacturer’s instructions. Both esophageal and stomach tissues were collected from the mouse of untreated and treated with RAN + lime with and without PRE for 180 and 260 days. For 8-OHdG measurement, DNA were extracted and digested with nuclease P1  and further treated with calf intestinal phosphatase  and denatured. 8-OHdG was quantified by quantitative ELISA assay in 96-well plate format. The quantity of 8-OHdG in the specimens were determined by comparing its absorbance with known 8-OHdG standard curve.Measurement of 8-Hydroxydeoxyguanosine (8-OHdG), a known marker of oxidative stress-mediated DNA damage, was estimated in DNA of esophageal and stomach cells with OxiSelectCells were collected from the inner layer of stomach from untreated mice (n = 10), RAN + lime and PRE + RAN + lime treated (n = 6 per point) for 180 and 260 days. Total RNA was extracted using an RNeasy Mini Kit . From 1 μg of total RNA, cDNA synthesis was performed using QuantiTect Reverse Transcription kit  according to the manufacturer’s protocol. For qPCR, cDNA was amplified using SYBER Green PCR mastermix according to the manufacture’s cycling condition for 40 cycle on a StepOnePlus amplification and detection system (Applied Biosystems). The primers of target genes used for this analysis were Mitotic arrest deficient 2 (Mad2) and Aurora A Kinase (AukA), and the primer sequences are listed in Table Cells were collected from the inner layer of esophagous and stomach from untreated (n = 5), RAN + lime and PRE + RAN + lime treated mice for 180 and 260 (n = 5 per point) days. In case of 180 days treatment with ECGU + RAN + lime 4 mice were used as a treated group. The cells were washed with ice-cold 0.1 M phosphate-buffered saline  and total protein was extracted with a lysis buffer containing 0.1% SDS, 2 mM EDTA, 1% NP-40, 1% sodium deoxycholate, 50 mM sodium fluoride, 100 U/ml aprotinin and 1 mM phenylmethylsulfonyl fluoride. After centrifugation, the cell lysate was collected and the protein concentration was determined using the bicinchoninic acid protein assay. Equal amount of protein (40 µg/well) were subjected to Novex Tris-Glycine 4–20% gradient gels and electrophoresis was performed in NuPAGE electrophoresis system . Then the proteins were transferred to a polyvinylidene difluoride membrane (Sigma) and probed with 1:1000 dilution of a mouse monoclonal antibody against p53 , Securin  and β-actin . Alkaline–phosphatase conjugated anti-mouse IgG  used as secondary antibodies and immunodetection was performed by treating the blot with the substrate solution of BCIP/NBT .46 and stained with hematoxylin and eosin47. Sections were then observed under a light microscope and photographed .Stomach tissue was collected from untreated (n = 5), RAN + lime treated and PRE + RAN + lime treated mice (n = 10 at each point) at 260 days. Tissue sections (5–7 μm) were processed for histological sectioning as per standard protocol48 (please see Supplementary section).For immunohistochemistry (IHC) analysis, a small part of stomach tissue were collected from untreated and treated (both RAN + Lime with and without PRE) mice which were used for histopathological evaluation and kept in formalin. The tissues were dehydrated, paraffin embedded and sectioned with a microtome (Leica). Briefly, after blocking for endogenous peroxidase activity, the sections were incubated with anti-p53  and anti-Securin  primary antibody. IHC analysis was performed with a Strept-Avidin Biotin Kit (Dako). The scoring of immunohistochemical stains in each specimen was determined using a histological score (H)The results are expressed as mean ± SEM for control and treated samples. Statistical analysis was performed by Student’s t-test with GraphPad Prism software 5.1. The values were considered statistically significant, if the p value was 0.05 or less.Methanolic extract of Potentilla fulgens root and its ethyl-acetate fraction delays the process of carcinogenesis in mice


The natural reservoir for all influenza A viruses (IAVs) is wild birds, particularly dabbling ducks. During the autumn, viral prevalence can be very high in dabbling ducks (> 30%) in the Northern Hemisphere, and individuals may be repeatedly infected. Transmission and infection is through the fecal–oral route, whereby birds shed viruses in feces and conspecifics are infected though feeding in virus-contaminated water. In this study we wanted to assess two alternative infection routes: cloacal drinking and preening. Using experimental infections, we assessed patterns of infection using a combination of virus shedding, as assessed by real-time PCR from cloacal swabs, and patterns of viral replication using virus-immunohistochemistry of gastrointestinal tissues. The cloacal drinking experiment consisted of two trials using cloacal inoculation at two different time points to account for age differences, as well as a trial whereby ducks were allowed to take up virus-laden water through the cloaca. All ducks became infected, and rather than the bursa of Fabricius being the main site of replication, the colon had the highest intensity of replication, as inferred through immunohistochemistry. In experiments assessing preening, feathers were contaminated with virus-laden water and all ducks became infected, regardless of whether they were kept individually or together. Further, naive contacts were infected by the individuals whose feathers were virus-contaminated. Overall, we reinforce that IAV transmission in dabbling ducks is multifactorial—if exposed to virus-contaminated water ducks may be infected through dabbling, preening of infected feathers, and cloacal drinking.The online version of this article (10.1186/s13567-018-0604-0) contains supplementary material, which is available to authorized users. Influenza A virus (IAV) is one of the most important viruses of the twentieth century . It is mHistomonas meleagridis causing Blackhead in poultry , at 24 h (1 dpi), and at 48 h (2 dpi). Cloacal samples were collected with a sterile tipped applicator and placed in virus transport media . Samples were stored at −80 °C within 4 h of collection. RNA was extracted using the MagNA Pure 96™ Nucleic Acid Purification System  and MagNA Pure 96 DNA and Viral Nucleic Acid Large Volume Kit (Roche) following manufacturer’s recommendations. Following extraction, samples were assayed by real time reverse transcriptase PCR (rRT-PCR) with the One Step RT-PCR Kit , targeting a short fragment of the IAV matrix gene  on a RocNecropsies were performed immediately following euthanasia. From each individual we collected gastrointestinal tract tissues: two seven-cm-long segments of jejunum (J1 and J2) and two seven-cm-long segments of ileum (I3 and I4) at intervals of approximately seven centimeters apart, colon (C), and cloacal bursa (B). All tissues were fixed in 10% neutral-buffered formalin (Sigma) for histopathology and IHC. Briefly, after fixation, samples were processed routinely, embedded in paraffin, and sectioned at 4–5 µm. Sections were processed for IHC using a commercial anti-influenza A nucleoprotein primary monoclonal antibody . Detailed histopathology and IHC methods are described in Bröjer et al. .Cells with distinct red staining in the nucleus or cytoplasm were identified as sites of virus replication, and tissues were considered positive even if only one or a few positive cells were present. The intensity and extension of the immunostaining was assessed semiquantitatively by the following scoring system: 0: no positive cells, 0.5: positive cells present, 1: mild, 2: moderate, 3: marked.p values of < 0.05 were taken to indicate a significant difference in the compared rates. Statistics were done using GraphPad Prism v.6.03 .Variation in Cq values across dpi within each trial were tested using a paired t-test and across trials within dpi were tested using ANOVA, following a test to ensure normal distribution of residuals. Two-way ANOVAs were not used due to small sample sizes. p = 0.5402; 6 months t = 1.3218, p = 0.2568; exposure t = 0.5021, p = 0.6501) or the preening trials . Despite apparent, but not significant variation at 1 dpi , all treatments had a similar range of Cq values at 2 dpi  , we assessed sections of the gastrointestinal tract using IHC for localization and intensity of infection. Patterns of viral replication suggest that cloacal inoculation does initiate infection, given that a large proportion of both 4 and 6 month old ducks had discernible positive cells Figure C and allOverall, when present (birds 4 and 5 months old) the bursa of Fabricius was free of IAV replication, except for scant replication observed in one individual. While bursal tissue could be identified in ducks at 4 and 5 months, 18/20 ducks that were 6 months  had no discernible bursal tissue , positive staining was found throughout the gastrointestinal tracts of all individuals Figure E, and foOne reason for the success of IAV in the waterfowl reservoir is its transmission efficiency, potentially driven in part by flexibility in mode of transmission and infection. In nature, water provides an important conduit, whereby ducks are infected through virus-contaminated water following dabbling. Indeed, uninfected ducks quickly become infected if sharing water with infected conspecifics, a premise which surveillance of sentinel ducks relies upon –35. The Cloacal drinking has been shown to be an infection route in birds for other diseases such as blackhead , and forThe results from the cloacal exposure trial were harder to interpret given that two of the individuals had very intense replication along the entire length of the gastrointestinal tract, markedly different from the cloacal inoculation trials. A possible explanation is that these birds became infected following preening or through droplets; that is, we were unable to remove all virus from feathers, feather oils in particular may have prevented out ability to remove all the virus effectively, or during the process of removal from inoculation boxes ducks created aerosols and were thus oesophagally infected. Alternatively, it could be due to individual differences in infection patterns or in susceptibility in the Mallards despite all individuals being from the same breeder . Given tAs hypothesized, following contamination of feathers with virus-laden water all ducks in all treatment groups were rapidly infected, with widespread IHC staining along the gastrointestinal tract. Birds in separate cages had similar patterns of infection to birds cohoused, and acted as individual replicates thus better eliminating the possibility of a contaminated drinking water source. All cohoused birds in the preening trial, including both the infected birds and the uninfected conspecifics had similar patterns, both in viral shedding and IHC patterns of infection. Patterns of infection in this experiment were very similar to those described in intra-oesophageal infection . The simWe demonstrate that IAV transmission in dabbling ducks is multifactorial. If exposed to virus-contaminated water, ducks may be infected through dabbling, preening of infected feathers, and cloacal drinking. Furthermore, even without a virus contaminated water source, contact ducks were infected by their conspecifics illustrating the role of direct contact, preening and allopreening. The findings are important to consider when conducting and interpreting surveillance and transmission studies of IAV in the natural host. Surveillance using feathers may, for example, be a useful tool in describing the viral diversity. This study highlights the ubiquitous nature and flexibility of IAV transmission in the Mallard host, consistent with a long history of host–pathogen co-evolution.Additional file 1.Schematic design for the purpose-built inoculation boxes for the cloacal drinking experiment.Additional file 2.Intensity of infection for all individuals in the study.


Despite the increasing number of studies on user experience (UX) and user interfaces (UI), few studies have examined emotional interaction between humans and deformable objects. In the current study, we investigated how the anthropomorphic design of a flexible display interacts with emotion. For 101 unique 3D images in which an object was bent at different axes, 281 participants were asked to report how strongly the object evoked five elemental emotions  in an online survey. People rated the object’s shape using three emotional categories: happiness, disgust–anger, and sadness–fear. It was also found that a combination of axis of bending  and convexity (bending convexly or concavely) predicted emotional valence, underpinning the anthropomorphic design of flexible displays. Our findings provide empirical evidence that axis of bending and convexity can be an important antecedent of emotional interaction with flexible objects, triggering at least three types of emotion in users. Shapes are closely related to emotion. In recent decades, there has been an increasing number of studies on user experience (UX) and user interface (UI) for deformable displays. With “anthropomorphism” as the philosophy of design, it is necessary to implement emotional interaction between humans and deformable displays to provide positive implementation of UX. Anthropomorphism, assigning human characteristics such as emotion to a non-human object, enables users to be familiar with the deformable display since people unconsciously derive emotional stability from things that are similar to themselves. Herein, this paper examines which functions would be appropriate to implement on these personified flexible devices in a theoretical framework, particularly focusing on interaction between emotional input and output.According to research, people unconsciously tend to be attracted to things that are similar to themselves . In the Figure 1).Currently, anthropomorphism is extensively researched, particularly in the field of humanoid robots or human–robot interaction (HRI). Indeed, in the field of HRI, it has widely been found that anthropomorphized technologies, in the form of both humans and animal creatures, increase social interaction and support emotional bonding with humans . For exaResearch on anthropomorphism in terms of interaction has not been limited to a humanlike appearance . For insAt the same time, recent studies have attempted to evaluate a novel expression method applying principles of animation to technology. For example, Flexible display, first conceptualized and prototyped by Xerox PARC  in 1974, refers to a dynamic display that can be forced out of shape. Display-related companies, such as Nokia , Sony , Samsung , and LG , have developed flexible displays . LeadingThere are two types of displays called flexible displays. The first is rigid and fixed in shape. Examples of this flexible display include the LG curved phone and Samsung Edge, which have glass material that works as a lower board and protects the display. The second type of flexible display is a dynamic display, which has an innovative form factor such that it can flexibly change its shape. For example, Samsung officially launched their flexible OLED display, called the Youm display, which was demonstrated at CES 2013 . AccordiBeyond stiff and brittle displays, there has been increasing research aiming to optimize UX and UI for flexible displays. Interaction methods of flexible displays have also been widely researched in recent years. According to Indeed, according to Extensive research has been conducted on the classification of emotion using facial expression and body shape . Among vFigure 2). For each axis, the device had one of three convexities: concave (bending forward), convex (bending backward), or flat (no bending). Strictly speaking, convex shape is often interpreted as either biconvex (both sides being curved outward) or plano-convex (single side being curved outward while the other side remains flat) while concave is interpreted as biconcave (both sides being curved inward) or plano-concave (single side being curved inward while the other side remains flat). However, as in In the current study, we used an online survey to investigate 281 users’ emotional evaluations regarding various shapes of an imaginary flexible device. The shape changes included folding, bending, rolling, pinching, zero-crossing, twisting, and crumpling. To generate these shapes, we computed all combinations of 60° bends in three horizontal, three vertical, three diagonal right, and three diagonal left axes , the max number of bendings was three for 2049 shapes. Then, technically impossible shapes were discarded. These shapes included (1) shapes bent on both vertical and horizontal axes without diagonal bendings, (2) shapes bent on either vertical or horizontal axes as well as on diagonal axes, and (3) shapes bent on any diagonal-right axes as well as on the middle diagonal-left axis and vice versa. After eliminating technically impossible shapes, 153 shapes remained. Finally, vertically mirrored duplicates – 52 shapes – were again eliminated. As a result, 101 unique shapes were generated. All data and materials are publicly available on the project page in the Open Science Framework1.To eliminate redundancy in combinations .These 101 possible 3D images of flexible displays were created using 3D Rhino. All the flexible displays featured an iPhone that was 67 mm × 138 mm × 7 mm. To remove the ambiguity that often occurs with a single viewpoint, each shape was rendered in two viewpoints – “distance = 266.12 mm, azimuth = -50°, Inclination = 15°” and “distance = 266.12 mm, azimuth = -132.53°, inclination = 53.22” – with the former as a main image positioned in the center and the latter as an additional image placed at the top right corner of the former . The block order was shuffled across participants, and shapes were presented in a random order in each block. The survey session took approximately 60–90 min. This study was carried out in accordance with the recommendations of the Institutional Review Board at Yonsei University. The protocol was approved by the Yonsei University Institutional Review Board. All participants, aged 18 and above, gave written informed consent in accordance with the Declaration of Helsinki. Participants received $10 in compensation for their participation and could quit anytime during the survey if they did not want to continue. No personal identifying information was collected.Table 1). Low reliability has often been pointed out as one of the major limitations of a web-based survey. To avoid this issue, we excluded outliers with three criteria. First, each emotion block included five checksum questions: five shapes were asked twice, randomly interleaved with the other 101 shapes for each emotion block. Participants were excluded if they gave different responses to the same question on two occasions (root mean squared error for 25 checksum questions was greater than 2.0). The first criterion detected 17 outliers. The second criterion was the max frequency in each block. Some participants could habitually give the same response for the most questions. Such outliers were detected by the max frequency; if a participant’s max frequency was greater than or equal to 101 (out of 106 questions including the checksum questions) in any emotional block , the participant was excluded. This criterion detected 25 outliers. With the second criterion, we could fail to find outliers who regularly changed answers . To exclude these outliers, we also computed a standard deviation over 530 answers  for each participant. If a participant’s SD was less than 1.0, we regarded the participant as an outlier. Following this rule, there were 58 outliers. In total, 87 outliers were detected and excluded in the following data analysis. Please note that some outliers belonged to two or more criteria. Responses for the checksum questions were also discarded in the analysis. Therefore, 101 responses for each block from the remaining 281 participants were analyzed in this study.A total of 368 participants volunteered to participate in the study. To obtain data from the general population, we attempted to assign an equal quota across age groups and gender. None of the age–gender groups exceeded 10 ± 2.5% , fear , and disgust  but a marginally significant negative correlation with anger . Among correlations between four negative emotions, sadness–fear  and anger–disgust  showed strong correlations.Although five elementary emotions on human facial expression have been identified , little z = 9.85, p < 0.001) or disgust . Fear also had significantly greater correlation with sadness than with anger  or disgust . That is, correlation between sadness and fear was significantly stronger than any other correlations involving sadness and fear. Likewise, anger had significantly greater correlation with disgust than with sadness  or fear . Disgust also had significantly greater correlation with anger than with sadness  or fear . These results suggest that correlation between anger and disgust was significantly stronger than any other correlations involving anger and disgust.To further investigate the relation between negative emotions, we tested the significance of the difference between correlation coefficients. As results, sadness had significantly greater correlation with fear than with anger  happiness, (2) sadness–fear, and (3) anger–disgust. In the following analysis, therefore, we will use three emotional categories. The sadness–fear score was calculated by averaging each participant’s sadness and fear ratings for each shape. Ratings for anger and disgust were also collapsed to produce the anger–disgust score.Figure 4, only a few shapes evoked high emotional responses in two categories, and none did in all three categories.To explore which shapes evoked strong emotional responses, we first selected shapes with an average rating higher than the mean + 1 SD of 101 rating scores  for each emotional category. With criteria of 3.585 (=3.317 + 0.26), 3.992 (=3.537 + 0.455), and 4.000 (=3.650 + 0.350), a total of 12, 11, and 15 shapes were selected for happiness, sadness–fear, and anger–disgust, respectively. As clearly shown in n = 281). Results suggested there were two factors whose eigenvalues were greater than 1.0 , accounting for 60.984% of the total variance. Factor loadings of highly happy shapes and their physical properties are summarized in Table 3, and images for shapes are shown in Figure 5. The table and figure clearly show the unique property of the first factor: a convex bending on horizontal axes. All these shapes had one or two convex bending(s) at the middle or the top horizontal axis, reminding us a laughing figure. The second factor was made up of combinations of convex and concave bendings on horizontal axes in a sandwiched manner  with either a simple (#46 and #48 with two bendings) or a complex shape (#80 or #87 with three bendings). These shapes reminded us a giggling figure.All high-happiness shapes had bendings on horizontal axes (mainly convex); none had a bending on vertical or diagonal axis. To further identify common factors of these shapes, we conducted an exploratory factor analysis with varimax rotation ; none had a bending on vertical or diagonal axis.n = 281). Results suggested a two-factor solution , explaining 73.307% of the total variance. Factor loadings of highly sad and fearful shapes and their physical properties are summarized in Table 4, and images for shapes are shown in Figure 6.We submitted all shapes to an exploratory factor analysis with varimax rotation . Indeed, shapes #80 and #87 evoked high happiness as well as high sad–fearful. These belonged to the second factors of both highly happy and highly sad and fearful shapes. It seemed that the common shapes and their properties  were perceived sometimes as giggling figures and sometimes as trembling figures.There were 15 shapes that participants rated highly angry and disgusting. In contrast to happy and sad–fearful emotional shapes, which had bendings on horizontal axes but no diagonal axis, all highly angry and disgusting shapes, except for #82, bendings on diagonal axes and no horizontal axis (neither concave nor convex). No emotional response was evoked by bending on vertical axis in any of the three emotional categories.n = 281) for high-anger–disgust shapes. Results showed that only the first factor had a dominant eigenvalue  compared to the remaining factors . The first factor accounted for 61.885% of the total variance. Physical properties of high-anger–disgust shapes are summarized in Table 5, and images for shapes are shown in Figure 7.We also conducted an exploratory factor analysis  at least once. Such a common property hinted to us that participants perceived these shapes as a figure twisting its shoulder.Figure 4, #82 belonged to high-anger–disgust shapes as well as high-sadness–fear shapes. It seemed that #82’s shape of shrinking its shoulder or body was often perceived as suggesting all negative emotional states.Shape #82 was again an exception for this property. As shown in In a series of studies, Ekman found that people – regardless of cultural background – categorized human facial expressions into five categories. In contrast, our study showed that people categorized objects’ shapes into three categories. Participants could not discriminate disgust from anger nor sadness from fear in our study. In some sense, it seems to be natural for sadness to be grouped with fear and anger with disgust. This is because, by nature, sadness and fear are directed inward (inside oneself) while anger and disgust are directed outward (toward an object outside of oneself).High correlation between disgust and anger was commonly found in previous literature exploring recognition of emotions . HoweverIn the field of robotics, the cognitive underpinnings of emotional interaction between human and anthropomorphized robotics is considered crucial since robots have “synthetic psychology,” a state of not possessing internal emotion regardless of external emotional expression . That isThis study found that the shape of the bended flexible display indicates a certain emotional expression, confirming the hypothesis that the anthropomorphic design of the flexible display would enable emotional interaction with the users. Since it is necessary to anthropomorphize the display using a simplistic pattern in the early stage of technical development, certain patterns that indicate emotion to users needed to be investigated. Here, based on the parameters that may underpin the emotional interaction of the flexible display, the axis of the bending and convexity of the curve were researched thoroughly. Through this empirical study, which examined how much the 3D-modeled image of a flexible display represents specific emotions to the participants, certain patterns have been found regarding the axis of bending and convexity. In terms of individual emotions, first, happiness was represented by the combination of a convexly curved display that has bendings on horizontal axes, significantly distinguishing itself from the other four emotions. Sadness and fear, which were highly correlated, had a concavely curved display that has bendings on horizontal axes. Last, anger and disgust, which also correlated with each other, had a curved display that has bendings on diagonal axes regardless of bending convexities.Broadly, taking the conventional recognition concept that happiness conveys positive emotion while the other basic expressions, sadness, anger, fear, and disgust, are negative, it can be said that convex shape triggered positive emotion while concave shape conveyed negative emotion to the participants. This perception can be analyzed via Russell’s Circumplex model  since emFrom this perspective, there is concern over misinterpretation of emotion expression since, regardless of subdivision of emotion group into categories, there are numerous ambiguous emotions. Indeed, according to The results in which the participants portrayed their emotional value on a flexible display align with the research conducted by Our findings suggested some basic parameters required for anthropomorphic design of an emotional object. First, bending in horizontal and diagonal axes should be available. Bending along the horizontal axis should be applicable to express happiness, sadness, and fear while bending along a diagonal axis would express anger and disgust. Interestingly, bending along the vertical axis was not a critical factor for triggering emotional interaction in the users. Second, both concave and convex bending should be feasible. The results revealed that convex bending is required to express happiness, concave bending for sadness and fear, and both concave and convex for anger and disgust.In our study, we explored the object characteristics that reflect specific emotion. This study provides insight for understanding the emotional interaction between a human and an anthropomorphic object. It has been empirically shown that there exists emotional interaction between human and flexible displays, and three emotional categories for anthropomorphic flexible display have been suggested . In thisFlexible displays have enormous potential, and many believe that they will be commercially viable in the near future. However, studies of the anthropomorphic design of flexible displays and their user interaction remain scarce. Our study aimed to provide significant information to researchers and designers who intend to develop emotionally interactive devices or designs.Although we successfully found factors influencing emotional interaction between humans and flexible objects, this study has a few limitations. First, the study was conducted with participants from a single cultural background. JML, JB, and DYJ conceived, designed, and conducted the study. All authors wrote, reviewed, and edited the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Ugandan Passiflora virus (UPV) sequenced from a symptomatic sample of KH7 passion fruit (Passiflora edulis Sims) variety. UPV had limited amino acid identity with other potyviruses known to cause passion fruit woodiness disease (PWD).Here, we present the first near-complete genome of  Ugandan Passiflora virus (UPV) sequenced from a symptomatic sample of KH7 passion fruit (Passiflora edulis Sims) variety. UPV had limited amino acid identity with other potyviruses known to cause passion fruit woodiness disease (PWD). The closest relationship (71.2% amino acid similarity) was with Bean common mosaic necrosis virus.Here, we present the first near-complete genome of  Passiflora edulis Sims)  was reported to cause PWD in Uganda (Passion fruit woodiness disease (PWD) is globally the most economically important disease of passion fruit (is Sims) . PWD is is Sims) –4. Recenn Uganda . To deven Uganda . Howevern Uganda . Here, wFive leaf samples were collected from a single plant belonging to KH7 variety and showing PWD-like symptoms. The KH7 plants were collected from a farmer’s field and maintained in an insect-proof screenhouse. Total RNA was extracted from the samples using a plant RNA miniprep kit (Zymo Research). An RNA quality check was done using Tris-acetate-EDTA (TAE)/formamide agarose gel electrophoresis  for quality control by first trimming with previously described quality parameter settings . The tri-2.2.29+ .De novo assembly yielded 43 contigs of >1,000 bp. The contig of interest was 9,670 nt with 8,100,407 reads mapping to it with a coverage of 126,047×. The 9,670-nt contig (referred to here as KH7-1) had a G+C content of 40.73%. A BLASTN search revealed that KH7-1 resembled a Ugandan Passiflora virus isolate, UGM-19a (GenBank accession number FJ896000), with 99.36% nt identity over 1,718 bp. Also, KH7-1 had a single ORF comprised of 10 mature proteins typical of potyviruses (Bean common mosaic necrosis virus isolates (ANO46357 and ARI46489). Although the UPV sequences currently present in GenBank originate solely from Uganda, we propose a more neutral name for this virus (Passiflora virus” (PV). This is because with increasing human movement and world trade involving plants/plant products, this virus might be found elsewhere  remained after trimming. yviruses , 12. Paiyviruses  showed lis virus , withoutMK110656. Raw data were deposited in the SRA under BioSample number SAMN10717829, which is part of BioProject number PRJNA513986.The sequence described here was deposited in GenBank under accession number


Several kinds of abnormalities or malformations affecting the female genital system of Dorylaimid nematodes have been repeatedly reported in longidorid forms, more occasionally in free-living taxa. These anomalies include total or partial duplication of the system, a didelphic-opisthodelphic condition, total or partial reduction of one genital branch, and the existence of two (or even three) vulvae , and might be, for instance, exposed to an extremely stressful environment during its development.Vulvaless or  elegans , Pristioacificus  and Osch tipulae , an indi


Cytorhabdovirus), tentatively named bean-associated cytorhabdovirus (BaCV). The BaCV genome presented all five open reading frames (ORFs) found in most rhabdoviruses: nucleoprotein (N) (ORF1) , phosphoprotein (P) (ORF2) (445 aa), matrix (M) (ORF4) (287 aa), glycoprotein (G) (ORF5) (520 aa), and an RNA-dependent RNA polymerase (L) (ORF6) (114 aa), as well as a putative movement protein (P3) (ORF3) (189 aa) and the hypothetical small protein P4. The predicted BaCV proteins were compared to homologous proteins from the closest cytorhabdoviruses, and a low level of sequence identity (15–39%) was observed. The phylogenetic analysis shows that BaCV clustered with yerba mate chlorosis-associated virus (YmCaV) and rice stripe mosaic virus (RSMV). Overall, our results provide strong evidence that BaCV is indeed a new virus species in the genus Cytorhabdovirus (family Rhabdoviridae), the first rhabdovirus to be identified infecting common bean.Using double-strand RNA (dsRNA) high-throughput sequencing, we identified five RNA viruses in a bean golden mosaic virus (BGMV)-resistant common bean transgenic line with symptoms of viral infection. Four of the identified viruses had already been described as infecting common bean  and one is a putative new plant rhabdovirus (genus  Phaseolus vulgaris L.) is an economically important leguminous crop grown worldwide. It is a staple food for human consumption and the most consumed legume worldwide. Brazil is the third largest producer and one of the major consumers of common beans in the world [Common bean , a seed-borne virus [Geminiviridae, genus Begomovirus) [Potyviridae, genus Potyvirus) [Betaflexiviridae, genus Carlavirus) [Different viruses have been reported infecting common beans. Some of them are asymptomatic, such as the double-strand RNA (dsRNA) virus Phaseolus vulgaris alphaendornavirus 1  ,5, bean tyvirus) , and cowlavirus) .Among bean-infecting viruses, BGMV is the primary limiting factor to bean production in Brazil and causes up to 100% crop losses ,8. To ovPlant infection by multiple viruses is common in nature and is frequently reported ,12. DuriIt was already known that BGMV-resistant transgenic bean lines are susceptible to CPMMV . HoweverAlphaendornavirus, Carlavirus, and Comovirus) and one newly discovered virus in the genus Cytorhabdovirus (family Rhabdoviridae), herein named bean-associated cytorhabdovirus (BaCV).More recently, transgenic lines in field experiments presented severe symptoms of mosaic, including leaf distortion, crinkling, and vein necrotic enations. These more severe symptoms could be the outcome of multiple virus infection in these plants. Diagnostic PCR tests were negative for the begomoviruses reported as infecting beans in Brazil , and Sida micrantha mosaic virus (SiMMV)). Therefore, to test for the presence of viruses with RNA genomes in the BGMV-resistant common bean transgenic line CNFCT16207, we performed high-throughput sequencing (HTS) of enriched dsRNA samples. We were able to identify five viruses, including four known viruses from distinct genera  and the extract was used for the mechanical inoculation of four common bean cv. Jalo Precoce and four transgenic BGMV-resistant line CNFCT16207 seedlings. Plants were kept in a greenhouse with temperatures varying between 24 °C and 36 °C. The inoculated plants were observed for up to 50 days and regularly checked for symptom development. Leaves were collected for dsRNA extraction.2O and stored at −20 °C. The presence of dsRNA was determined by agarose gel electrophoresis.dsRNA extraction was done based on modified protocols of Valverde et al.  and Castn = 12) was pooled in a single tube  and sequenced using the MiSeq sequencing platform  at the Universidade Católica de Brasília (UCB). Total dsRNA was converted to cDNA using random hexamers, fragmented into libraries using the transposon-based Nextera™ DNA Sample Prep Kit and sequenced using MiSeq Reagent Kits v2 (2 × 150 bp) . The raw reads were quality trimmed and assembled de novo using CLC Genomics Workbench version 6.3. The resulting contigs were compared to the complete viral RefSeq database  using Blastx [dsRNA from all samples . The reaction mixes consisted of 1 to 2.5 µL of dsRNA, 6.3 µL of 2× reaction mix, 1 µL of each forward and reverse primers (10 μM), and 0.4 µL of SuperScript® III RT/Platinum® Taq mix in a final volume of 12.5 µL. The cycling conditions were 1 cycle for cDNA synthesis at 53 °C for 20 min; 1 cycle for denaturation at 94 °C for 2 min 30 s; 35 cycles for PCR amplification at 94 °C for 20 s, specific primer Tm °C . Four clones for each virus-derived fragment were Sanger-sequenced at Macrogen Inc. .Based on the assembled contigs, specific primers were designed for each virus identified . Each saer Tm °C  for 35 sEscherichia coli poly(A) polymerase ; then, the cDNA was synthesized using an oligo(dT) primer with an anchor sequence, M10PacIT50VN . Four clones of each fragment were randomly selected and Sanger-sequenced at Macrogen Inc To determine the 5′- and 3′-terminal sequences of BaCV genome, rapid amplification of cDNA ends (RACE) was performed according to the method described previously ,19. The acIT50VN . PCR reaacIT50VN . The 5′ The viral sequences identified were aligned with their corresponding homologs available in GenBank using MAFFT (Multiple Alignment using Fast Fourier Transform) . PhylogeCytorhabdovirus).High-throughput sequencing technology and bioinformatics tools have improved the discovery and detection of new viruses. Recently, HTS has been used to describe bean-infecting viruses, such as CPMMV, PvEV-1, PvEV-2, and a newly described Phaseolus vulgaris alphaendornavirus 3 (PvEV-3) ,26. HereThe presence of these viruses was confirmed by RT-PCR  and SangBetaflexiviridae, genus Carlavirus) and named CPMMV:BR:GO:14 (Glycine max) [Desmodium tortuosum and a sequence from whitefly vector-enabled metagenomic (VEM) data from Florida (USA) [A contig of 8194 nucleotides (nt)  was identified as CPMMV  , and in ine max) , with anda (USA) . CPMMV:BAlphaendornavirus genus (family Endornaviridae) were also identified in our analyses: PvEV-1, and PvEV-2  , were identified as the bipartite bean rugose mosaic virus  . Both gemovirus) . PairwisCytorhabdovirus. The contig presented the typical genomic organization of the viruses members of the family Rhabdoviridae, including a 3′ leader followed by seven open reading frames (ORFs) and a 5′ trailer sequence [A contig sequence of 13,448 nt (∼26× coverage) showed low similarity (39% aa identity) with the northern cereal mosaic virus (NCMV) RdRP protein (NP_597914.1), a member of the genus sequence   (ORF1) (451 aa), phosphoprotein (P) (ORF2) (445 aa), matrix (M) (ORF5) (287 aa), glycoprotein (G) (ORF6) (520 aa), and an RNA-dependent RNA polymerase (L) (ORF7) (2114 aa), as well as a putative movement protein (P3) (ORF3) (189 aa) and a hypothetical protein P4  , which ibdovirus . The prebdovirus . Consecubdovirus . No TURBCytorhabdovirus (family Rhabdoviridae).Moreover, the predicted BaCV proteins were compared to homologous proteins from the closest cytorhabdoviruses, revealing a low level of sequence identity, ranging from 15% to 39% . The nucIn summary, HTS was instrumental in uncovering RNA viruses infecting BGMV-resistant transgenic plants presenting severe symptoms in the field. Our findings extend the knowledge of bean-infecting viruses in Brazil and warrants further studies on the biology, insect transmission, and distribution of BaCV, the first cytorhabdovirus to be identified infecting common beans.


The adsorption capacities of the beads were evaluated under both batch and dynamic conditions. The adsorption capacity was found to be ~29 mg/g of adsorbent at 298 K when the equilibrium concentration of strontium in the solution was 925 mg/L at pH 6.5. The X-ray photoelectron spectroscopy (XPS) data suggest that strontium uptake by the beads occurs mainly through an ion-exchange process. Kinetic data indicate that the sorption of strontium onto the beads follows anomalous diffusion. Thermodynamic data suggest that the ion-exchange of Sr2+ on the bead surface was feasible, spontaneous and endothermic in nature.Fuller’s earth spherical beads using chitosan as a binder were prepared for the removal of strontium ions from aqueous solution. The adsorbents were characterized by scanning electron microscopy (SEM) and transmission electron microscopy (TEM), which revealed the porous nature of the beads. The Brunauer–Emmett–Teller (BET) surface area of the beads was found to be 48.5 m Strontium is one of the major radionuclides that are present in radioactive wastewater, which is, in general produced during the nuclear fission process. Strontium is a pure beta emitter of 0.546 MeV with a half-life of 28.8 years and is commonly found in many groundwater systems migrating from historic nuclear waste storage sites ,2. Once olutions ,12,13. Aolutions . The revolutions . The aff2+ adsorbs to a wide range of aluminosilicates, Fe oxides and other soil minerals via weakly bound surface complexes [In general, the radionuclide Sromplexes . Severalomplexes ,18,19,20In recent years, the natural clay mineral based low-cost adsorbents have become attractive again for water purification and radioactive waste management . For exaSeveral studies ,23 reporFuller’s earth was obtained from Fisher Scientific Co., Hampton, NH, USA. The CF adsorbent was prepared as spherical beads using chitosan as a binder. The chitosan used in this study was 75 to 85% deacetylated and had a molecular weight of ~190,000–310,000 as determined from viscosity data by Aldrich Chemical Corporation. All chemicals used in this study were of analytical grade. A stock solution containing 1000 mg/L of strontium was prepared by dissolving strontium chloride in ultrapure water. The working solutions of various strontium concentrations were obtained by diluting the stock solution with ultrapure water.Fuller’s earth powder of 35 mesh size was first soaked in 0.2 mol/L oxalic acid for 4 h. This helped to introduce various functional groups into Fuller’s earth, which in turn helped the adhesion of chitosan onto Fuller’s earth later during the bead making process. It was then washed with distilled water and dried in an oven at 60 °C for 12 h. Although washing with distilled water was not necessary, it was done to maintain a desired ratio of oxalic acid to chitosan which was found to be important for the performance of the beads as adsorbent. A higher concentration of oxalic acid would require a greater concentration of NaOH as well as a greater amount of water during the bead making process. An amount of 60 g of acid-washed Fuller’s earth was mixed with 30 g of chitosan flakes in a beaker with 1 L of 0.2 mol/L oxalic acid. The mixture was stirred for 4 h while heating at 313–323 K (40–50 °C) to obtain a homogeneous mixture. The spherical CF beads were prepared by drop-wise addition of the mixture into a 0.7 mol/L NaOH precipitation bath. The purpose of adding acidic Fuller’s earth-chitosan mixture to the NaOH solution was to assist rapid neutralization of oxalic acid so that the spherical shape of the bead could be retained. The CF beads were washed with deionized water to a neutral pH and oven dried for subsequent use.The surface area, pore volume, and pore diameter of the CF bead were evaluated by nitrogen adsorption/desorption at 77 K using the Brunauer–Emmett–Teller (BET) method . The adsorbents were outgassed at 378 K under a vacuum of <10 μm Hg for a minimum period of 24 h. The equilibrium nitrogen adsorption data and the liquid nitrogen temperature in the relative pressure range of 0.05 to 0.30 atm (0.74 to 4.41 psia) were used to calculate the surface area. Scanning electron microscopy (SEM) (AMRAY 1600T) was used to investigate the surface morphology of the CF beads. The SEM micrographs were taken using backscatter electrons with an accelerating potential of 20 keV. The surface morphology of the CF bead was also investigated using a Philips EM 430 for scanning transmission electron microscopy (STEM). The microtome technique was used to prepare the sample for TEM analysis. X-ray photoelectron spectroscopy (XPS) is extremely useful for studying the valence shell and for chemical analysis of the variety of compounds. The XPS analysis of the CF bead before and after exposure to strontium ion was performed using a KRATOS model AXIS 165XPS spectrometer, with non-monochromatic magnesium X-rays (hν = 1253.6 eV) used as the excitation source at a power of 240 W. The spectrometer is equipped with an 8-channel hemispherical detector, and the pass energy of 5 to 160 eV was used during the analysis of samples. Each sample was exposed to x-rays for the same period of time and intensity.The pH of the solution affects the degree of ionization, the surface charge, and the speciation of strontium, all of which can impact the adsorption mechanism and the uptake capacity of the adsorbent. The effect of pH on strontium removal from aqueous solutions was determined over the pH range of 2 to 11. The pH studies were carried out by exposing approximately 0.25 g of the CF beads to 100 mL of 20 mg strontium/L solution at 298 K. The pH of the solutions was adjusted by adding either 0.1 mol/L hydrochloric acid or 0.1 mol/L sodium hydroxide. The pH of the solutions was measured using a pH meter equipped with a glass electrode .Equilibrium batch adsorption studies were carried out by exposing the beads to different concentrations of aqueous solutions of strontium in 125 mL Erlenmeyer flasks to a pre-determined pH and temperature. Approximately 0.25 g of the beads was added to 100 mL of solution. Such a quantity of the beads and solution ensured that an equilibrium condition was reached, i.e., all of the strontium was not adsorbed by the beads, which would have made it difficult to determine the equilibrium point. The flasks were placed in a constant temperature shaker bath for a specific time period. Following the exposure of the CF beads to strontium, the samples were collected at predetermined time intervals. The solutions were filtered and the filtrates were analyzed for strontium by an atomic absorption spectrometer . The adsorption isotherm at a particular temperature was obtained by varying the initial concentration of strontium ions. The amount of strontium adsorbed per unit mass of adsorbent  [2+ and SrOH+ species in a wide pH range [+ is the main form of Sr2+ at pH > 12.8 [2+) with the release of H+ ions. Since Fuller’s earth mainly consists of hydrated aluminum silicate, the formation of AlO− and SiO− on the CF bead surface is also therefore possible due to the deprotonation of the surface hydroxyl group. The H+ ion could be released from the CF bead surface into the solution as a result of ion exchange. Therefore, the Sr2+ adsorption may involve surface hydroxyls of the CF beads according to the ion-exchange mechanism [The effect of pH on strontium uptake by the CF beads and Fuller’s earth is shown in nitrate) . The surpH range , but SrOH > 12.8 . From thechanism ,31:(2)−22+ uptake by the CF beads at various temperatures were also performed at pH 6.5. The equilibrium adsorption data in the temperature range of 293–308 K appear to be of a Type I shape, suggesting a monolayer adsorption of strontium onto the CF bead  to 11.36 mmol/L (1000 mg/L). It was observed that the adsorption rate was time dependent. Almost 70% of strontium was adsorbed during the first 4 h of a run, and then the equilibrium was attained monotonically at 10 h in most of the runs. An exposure time of 24 h was used during batch studies to ensure that equilibrium was attained . The equ CF bead .2+ uptake by the CF beads were fitted to the Langmuir isotherm equation. In general, the Langmuir isotherm provides a reasonable description of Type I systems and is often justified based on its ability to fit equilibrium data. The Langmuir equation can be expressed as:The equilibrium data for Sr2+ to the active sites on the CF bead, it was exposed to 100 mL of a 1.14 mmol/L strontium solution at pH 6.5. After 24 h of exposure, the beads were removed from the solution and dried at room temperature. The XPS analysis of the beads before and after the adsorption of strontium was used to gain a better understanding of the adsorption sites onto which strontium was adsorbed. To understand the binding of SrThe magnitude of the C-1s binding energy changed slightly only when the CF beads were exposed to strontium solution, whereas no significant change was observed for the O-1s and N-1s peaks a–c. The 2+ from the CF bead may take place by Sr2+ during the adsorption process; therefore, Sr2+ uptake by the CF beads may occur following an ion-exchange process. Based on the sorption kinetic and XPS analysis, it was hypothesized that the adsorption mechanism of Sr2+ onto the CF beads surface follows an ion-exchange mechanism. The reaction is given as follows:In comparison with the XPS , the binThe Gibbs free energy change equation is also used to gain insight into the thermodynamic nature of the strontium sorption process onto the CF beads. The Gibbs free energy change, ∆G, can be used to identify whether the chemical reaction that occurs during adsorption process is a spontaneous reaction or not. Therefore, it is considered as an important criterion for spontaneity. The Gibbs free energy equation for an adsorption process can be expressed as:exposure . TherefoG with temperature indicates more efficient adsorption at a higher temperature; therefore, it requires energy to carry out the sorption reaction process. The increase in perature . The risperature  as well 2+ on the CF bead was found to be 8.287 kJ/mol  is obtained by plotting −1·min) can be obtained by plotting Adsorption kinetics data explain the rate of the adsorption of metal ions on the solid phase. For a batch contact process, the rate of sorption of strontium onto the CF bead surface is proportional to the amount of strontium uptake from the solution phase. A pseudo-first-order equation can be employed to describe reactions, whereas the pseudo-second-order equation affords the sorption capacity on the solid phase. Therefore, a pseudo-first-order or a pseudo-second-order model equation can be used to explain the kinetics of strontium sorption onto the CF beads. Linear forms of the first-order and second-order models, based on the adsorption equilibrium capacity, are as follows: process .2+ uptake on the CF bead. This equation was first proposed by Roginsky and Zeldovich; but it is commonly known as the Elovich equation and is extensively applied to chemisorption data [The Elovich equation was fitted to experimental data in order to investigate Srion data . The gen2 > 96) value suggesting the applicability of Elovich equation.Integrating Equation 12 for the boundary conditions Sorption kinetics of strontium on the CF beads using 220 mg/L (2.5 mM/L) strontium solution was evaluated at pH ~6.5 at different temperatures (293–308 K). The exposure time of 24 h was used during batch studies to ensure that equilibrium was attained. The data were fitted to the pseudo-first-order, pseudo-second-order, and Elovich equation. It was observed that the kinetic constants, perature . With a 2+ into the CF bead [In addition, the Weber–Morris model was also used to investigate the rate constant of the intra-particle transport of Sr CF bead .(14)qt=1/2) and In Equation (14), eratures . This inR22 > 99 . TherefoThe strontium sorption kinetics was further analyzed for the CF bead with radius, equation :(15)F=1−CF beads . To inveTaking −7 to 1.94 × 10−7 cm2/s for the particle diffusion coefficient for the temperature range of 293 K to 308 K.The diffusion co-efficient was estimated from the slope  and mean2+ on the CF bead was feasible and spontaneous. The activation energy data according to the Arrhenius Equation suggest that the adsorption process was endothermic, and the process was governed by the interaction of physical nature. It is important to note that the regeneration of the column with the acidic solution causes swelling of the bead which may create hydrodynamic pressure build-up in the column. Future studies need to consider a suitable desorption solution in order to minimize swelling of the bead in the column.Fuller’s Earth was coated successfully by chitosan and prepared as spherical beads with diameters of approximately 2 mm. The SEM micrograph reveals the heterogeneously porous nature of the beads. The equilibrium adsorption capacity of strontium was found to be dependent on solution pH. A one-site Langmuir model provided the best fit to the adsorption data for strontium. The maximum adsorption capacity of strontium onto the CF beads at a pH of 6.5 was found to be ~30 mg/g at 298 K. XPS analysis indicated that the constituent of Fuller’s earth, for example calcium, is the active metal exchange site for the CF beads. The ion-exchange reaction in fact differs from adsorption. Since nearly every ion-exchange process is accompanied by adsorption and desorption, adsorption is sometimes indistinguishable from ion exchange. It is evident from the surface charge and XPS analysis that strontium is removed from the aqueous solution by the CF bead through an ion-exchange reaction. The adsorption reaction may also occur at the hydrated oxide of the surface as the adsorption of strontium onto the CF bead was found to be pH dependent. The kinetic data suggest that the ion-exchange process follows anomalous diffusion. The negative ∆G value shows that the ion exchange of Sr