In the absence of definitive diagnosis, healthcare providers are likely to prescribe empirical antibacterials to those who test negative for malaria. This problem is of critical importance in Southern Asia (SA) and South-eastern Asia (SEA) where high levels of antimicrobial consumption and high prevalence of antimicrobial resistance have been reported. To improve management and guide further diagnostic test development, better understanding is needed of the true causative agents of fever and their geographical variability.We conducted a systematic review of published literature (1980–2015) to characterise the spectrum of pathogens causing non-malarial febrile illness in SA and SEA. We searched six databases in English and French languages: MEDLINE, EMBASE, Global Health (CABI) database, WHO Global Health Library, PASCAL, and Bulletin de la Société Française de Parasitologie (BDSP). Selection criteria included reporting on an infection or infections with a confirmed diagnosis, defined as pathogens detected in or cultured from samples from normally sterile sites, or serological evidence of current or past infection.Salmonella Typhi, Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, and coagulase negative Staphylococcus were the commonly reported bacteria, while dengue virus, chikungunya virus, Japanese encephalitis virus, hepatitis B virus, and hepatitis C virus were common viral pathogens reported. Reports of rarely reported or emerging pathogens included a case report of Borrelia burgdorferi (Lyme disease) in India in 2010 and reports of Nipah virus in Singapore and India.A total of 29,558 records from 19 countries in SA and SEA were screened, of which 2410 (8.1%) met the selection criteria. Bacterial aetiologies were reported in 1235 (51.2%) articles, viral in 846 (35.1%), parasitic in 132 (5.5%), and fungal in 54 (2.2%), and 143 (6.0%) articles reported more than one pathogen group. In descending order of frequency, This review summarises the reported non-malaria pathogens that may cause febrile illness in SA and SEA. The findings emphasise the need of standardising the reporting of aetiological studies to develop effective, evidence-based fever management and improved surveillance. Research and development of diagnostic tools would benefit from up-to-date epidemiological reporting of the regional diversities of non-malaria fever aetiologies.CRD42016049281PROSPERO registration,  While a global decline in malaria burden has been reported in the last 15 years, Asia has observed the sharpest decrease . The mosFever is one of the commonest reasons to seek medical attention in this region . HoweverThus, finding available and pertinent evidence to compensate for the lack of aetiological knowledge of NMFIs will aid in enhancing surveillance strategies, fever diagnostics, and effective fever management and contribute to antimicrobial stewardship efforts in this densely populated region. In line with this, an initial effort was made in 2012 to map the regional distribution and abundance of the pathogens in the Mekong sub-region and this project expands that work .A major challenge for assimilation of available evidence on NMFI is the dearth of epidemiologically sound, methodologically rigorous, and standardised evidence. This precludes us from reliably assessing the distribution of prevalent fever-causing agents in the SA and SEA regions , 18. In This review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement . The stuA systematic literature search was carried out in six libraries: MEDLINE, EMBASE, WHO Global Health Library (SEARO and WPRO files), Global Health (CABI), Banque de Données Santé Publique (BDSP), and PASCAL to identify publications from 1980 through 2015. A broad search string was employed which consisted of Medical Subject Headings (MeSH) and free text terms with no restriction on study design  and then full-text screening (stage II). Only articles meeting the pre-specified inclusion and exclusion criteria were included. Non-malarial febrile illness: a systematic review of published aetiological studies and case reports from Africa, 1980-2015. Two reviewers assessed the articles for SA (PS) and SEA (CON) independently—with each article assessed only once by one of the two reviewers. The PRISMA flow diagram is presented in Fig. Data from articles deemed eligible for the review were extracted into a bespoke online data extraction form built for the purpose of this review. The extracted variables included the following bibliographic metrics: study title, names of the first and second author, year of publication, Uniform Resource Locator (URL), and the digital object identifier (DOI). Information on the following study meta-data was extracted: name of the study site, latitude and longitude of the study site(s), study start year, study end year, and range of age groups included. The type of sample, number of individuals tested, number of cases testing positive for an organism, and laboratory method/s used were recorded for the reported pathogens.Each of the reports identified in the review was classified into one of three groups: (i) case report/case series, (ii) seroprevalence studies, and (iii) fever series. A fever series was defined as studies where the number of participants tested was clearly stated together with the number of participants testing positive for a given pathogen with an accurate diagnostic test for pathogen identification (culture or molecular methods). Seroprevalence studies were defined as studies that reported the number of individuals testing positive for a serological test along with the total number of tested individuals. The seroprevalence studies were further classified into those carried out in symptomatic patients to diagnose acute infections and the ones carried out in asymptomatic individuals to measure past exposure or infection.Infections were categorised as bacterial, fungal, parasitic, or viral and were sub-categorised using an epidemiological definition based on the predominant mode of transmission for the pathogens into mutually exclusive groups as contact , vector-borne, airborne, and food- and/or water-borne .Study populations were grouped into four mutually exclusive categories: neonates (aged < 28 days), infants (1 to < 12 months), children (1 to < 13 years), and older individuals (≥ 13 years). If a study reported any participants from each age group, then they were grouped as participants of “all ages”. Countries were classified into sub-regions (“Southern Asia” and “South-eastern Asia”) according to United Nations designation of areas and regions .The unit of analysis was a published article for each pathogen. Articles reporting a given pathogen were categorised by the UN sub-regions, patient age group of tested individuals, pathogen class , and predominant epidemiologic mode of transmission. The heterogeneity of study design, pathogens sought, laboratory methods, reporting, and limitations in data extraction precluded meta-analysis or estimation of pathogen prevalence. Multiple articles reporting different pathogens from the same study were treated as two separate articles. All analyses were carried out using R software version 3.6.0 , and graphical presentations were done using ggplot2 library .The currently available tools for assessing the quality and risk of bias were not applicable to our review design , 24. We The database search identified a total of 29,558 articles, with 20,442 and 9116 articles for SA and SEA, respectively. Of these, 2410 unique articles were selected for the final synthesis  followed by Pakistan (n = 194), Nepal (n = 102), Bangladesh (n = 92), and Sri Lanka (n = 80). Most reports from SEA were from Thailand (n = 301), followed by Malaysia (n = 154), Singapore (n = 98), Indonesia (n = 58), and Vietnam (n = 52). There were fewer than 10 reports from each of Afghanistan, Bhutan, Brunei Darussalam, Maldives, Myanmar, and Timor-Leste.Data were collected from 2075 unique study sites, 1880 (90.6%) of which were within a radius of 50 km of the nearest city Fig. . In SA, Studies reporting on neonates were found in 176 (7.3%) articles, infants in 47 (2.0%), children aged 1 to < 13 years in 408 (16.9%), and older children and adults (≥ 13 years) in 764 (31.7%). There were 757 (31.4%) articles that included participants of all ages, while the age group studied was not reported in 258 (10.7%) articles  reports included in this review. Cerebrospinal fluid (CSF) samples were analysed in 101 (4.2%) articles; both CSF and blood in 60 (2.5%); bone marrow, joint, lymph, or liver aspirates in 63 (2.6%); peritoneal, pericardial, or pleural fluids in 23 (1.0%); spleen samples in 10 (0.4%); and vitreous humour samples in 10 (0.4%). Multiple sample sources were analysed in 73 (3.0%) articles, and the specimen analysed was not specified in two (0.1%)  articles, serological assays in 340 (24.7%), PCR in 30 (2.2%), and microscopy and staining in seven (0.5%), and multiple diagnostic methods were reported in 89 (5.8%) articles. For viruses, 680 (75.5%) articles reported serological testing, and 90 10.0%) PCR methods, 14 (1.6%) culture methods, and multiple diagnostic methods were reported in 117 (13.0%) articles. Fungal infections were identified using culture methods in 125 (87.4%) articles while parasites were detected using microscopy methods in 70 (51.1%) articles and serological tests in 39 (5.6%) articles. Further details are presented in Supplemental file 0.0% PCR Bacterial infections were reported in 1235 (51.2%) articles, viral infections in 846 (35.1%), parasitic infections in 132 (5.5%), and fungal infections in 54 (2.2%), and 143 (6.0%) articles reported more than one pathogen group , Escherichia coli (n = 270), Staphylococcus aureus (n = 255), Klebsiella pneumoniae (n = 169), and coagulase negative Staphylococci (n = 160) were the top five commonly reported bacteria  was the most commonly reported bacterium in Southern Asia  was the most commonly reported  followed by Rickettsia typhi (n = 48); Rickettsia, spotted fever group (n = 37); and Rickettsia spp. (n = 34)  articles from Southern Asia and 47 (6.4%) from South-eastern Asia. There was a single report of Borrelia burgdorferi from India in 2010.Salmonella Typhi (n = 285 articles) was the most common cause of food- and/or water-borne bacterial infections, followed by Leptospira spp. (n = 125) and Burkholderia pseudomallei (n = 108). Among neonates, Salmonella spp. were reported in seven articles followed by Burkholderia pseudomallei (n = 6) and Listeria monocytogenes (n = 4); this was similar among children aged 1 to < 13 years. Among adolescent children and adults (≥ 13 years), Burkholderia pseudomallei (n = 58), Leptospira spp. (n = 44), and Salmonella Typhi (n = 37) were the most common reported pathogens. Burkholderia pseudomallei was reported in 86 articles from SEA and 22 from SA. Streptococcus suis was reported in SEA from Cambodia , Lao PDR , Vietnam , and Thailand .Escherichia coli (n = 270 articles), Staphylococcus aureus (n = 255), and Klebsiella pneumoniae (n = 169) were the most commonly identified bacteria which spread through contact. Among neonates, Klebsiella pneumoniae was reported in 60 articles, while among children aged 1 to < 13 years, Streptococcus pneumoniae (n = 54), Staphylococcus aureus (n = 50), Escherichia coli (n = 45), Haemophilus influenzae (n = 31), and Pseudomonas aeruginosa (n = 27) were the top five commonly reported bacteria  was the only description of airborne bacterial infection, while there were no reports among older children.n = 593) followed by chikungunya virus (n = 102), Japanese encephalitis virus (n = 71), hepatitis B virus (n = 35), and hepatitis C virus (n = 33) , herpes simplex virus (n = 3), chikungunya virus (n = 2), enterovirus (n = 2), Japanese encephalitis virus (n = 1), and flavivirus (n = 1). Among infants, dengue virus (n = 8), rubella virus (n = 2), and an article each reported: chikungunya virus, Coxsackie B virus, hepatitis B virus, human herpesvirus 6, and B19 virus. Among children, dengue virus (n = 118), Japanese encephalitis virus (n = 28), measles virus (n = 9), hepatitis A virus (n = 7), and human herpes simplex virus (n = 7) were the top five commonly reported viruses. Among adults, dengue virus (n = 184), chikungunya virus (n = 28), Crimean-Congo haemorrhagic fever virus (n = 16), hepatitis B virus (n = 15), and Japanese encephalitis virus (n = 12) were the top five reported viruses. The age-stratified distribution of the reports on viral infections is presented in Supplemental file Dengue virus infection was the most commonly reported viral infection (33) Fig. . Among nirus n = 02, Japann = 593) followed by chikungunya virus (n = 102), Japanese encephalitis virus (n = 71), Crimean-Congo haemorrhagic fever virus (n = 31), and West Nile virus (n = 17). Zika virus was reported in an article each from Indonesia (1981), Pakistan (1983), and the Philippines (2015).Dengue virus was the most commonly reported arbovirus (Hepatitis A virus was reported in 17 articles and hepatitis E virus in 16. Human poliovirus was reported in an article from India (1991), and there was one report describing Coxsackie B virus from Thailand (1991). There were no other descriptions of food- and/or water-borne viral infections.n = 14), measles virus (n = 13), hantavirus (n = 7), human respiratory syncytial virus (n = 2 articles), and Sin Nombre virus (n = 1) were the airborne viral infections reported. Hantavirus was reported in an article each from Cambodia, Indonesia, and Thailand and in two articles each from India and Singapore.Hantaan virus (n = 35), hepatitis C virus (n = 33), rubella virus (n = 16), and human herpes simplex virus (n = 12) were the most commonly reported viruses spreading through contact. Nipah virus was reported from Malaysia , Singapore , India , and Bangladesh .Hepatitis B virus (Leishmania donovani was the most reported parasite (n = 50 articles) followed by Leishmania spp. (n = 31), Wuchereria bancrofti (n = 7), Naegleria fowleri (n = 6), and Trichinella spp. (n = 6). All of the reported Leishmania donovani were from Southern Asia  were the most commonly reported, with 31 articles describing Candida albicans, followed by Cryptococcus neoformans (n = 24) and Candida tropicalis (n = 12)  across the time periods and regions  considered as emerging or of regional interest is presented in Fig. Streptococcus suis was reported only in SEA, from Cambodia , Lao PDR , Vietnam , and Thailand . Nipah virus was reported from Malaysia , Singapore , India , and Bangladesh .The distribution of the study sites for some of the pathogens  and low in 1229 (51.0%). Of 2075 studies where it could be ascertained, 1880 (90.6%) were from urban settings or located within a radius of 50 km of the capital city or another major city  or the national capital. The selection of study sites is often influenced by academic institutions, logistical considerations, and funding availability. This limits the generalizability of the results to the whole country or region and clearly represents a reporting bias toward urban settings, in these regions with a predominantly rural population, as has been reported earlier in a systematic review aimed at characterising febrile illness in the Greater Mekong sub-region . SimilarOur review shares many of the limitations discussed in the preceding articles in this series . As in tThere were some additional limitations specific to this review. We inadvertently failed to include studies from Iran, formally recognised as a part of Southern Asia by the UN. We also searched for articles only in a limited number of languages and did not search for publications in local languages. The articles were assessed by one independent reviewer owing to the broad scale of this review. Finally, a large proportion of the studies included in this review were considered to be at high risk of bias and the results of this systematic review should be interpreted taking this into consideration  without omitting malaria are life threatening. Southern Asia and South-eastern Asia are major hotspots for infectious diseases with almost a third of humanity being at risk [Some of these infections can be relatively benign and self-limiting  while others (e.g.  at risk . A non-l at risk –42. ThisThe map of pathogen distributions presented in this review provides regional data that can form a basis for targeted development of diagnostic tools and fever case management strategies in SA and SAE.Additional file 1.Additional file 2.


The overall shape features that emerge from combinations of shape elements, such as “complexity” and “order”, are important in designing shapes of industrial products. However, controlling the features of shapes is difficult and depends on the experience and intuition of designers. Among these features, “complexity” is said to have an influence on the “beauty” and “preference” of shapes. This research proposed a Gaussian curvature entropy as a “complexity” index of a curved surface shape. The proposed index is calculated based on Gaussian curvature, which is obtained by the sampling and quantization of a curved surface shape and validated by the sensory evaluation experiment while using two types of sample shapes. The result indicates the correspondence of the index to perceived “complexity” (the determination coefficient is greater than 0.8). Additionally, this research constructed a shape generation method that was based on the index as a car design supporting apparatus, in which the designers can refer many shapes generated by controlling “complexity”. The applicability of the proposed method was confirmed by the experiment while using the generated shapes. In designing shapes of products, the overall shape features that emerge due to combinations of shape elements, such as points and lines, are important . These fFrom the above, since “complexity” is said to have an influence on “beauty” and “preference” of shapes, there are many studies regarding the quantification of “complexity” of curves and curved surface shapes, which are important in designing shapes. For example, Vitz defined the number of lines as “complexity” and analyzed the relationship between “preference” and the number of lines in black line drawings that are generated by the random-walk method . Backes This research aims to propose a “complexity” index of curved surface shapes while using Gaussian curvature entropy and construct a curved surface shape generation method that is based on the index to support the product design. This paper is organized, as follows. Chapter 2 presents the proposal of Gaussian curvature entropy as a “complexity” index of curved surface shapes. Chapter 3 describes the construction of a curved surface shape generation method that is based on the index and its validation experiment. Chapter 4 summarizes the achievements and future tasks.A conventional “complexity” index of curved surface shapes fails to evaluate shapes with small concavities, as is mentioned in the previous section. This research aims to solve the problem by adopting the information entropy, which evaluates “complexity” to evaluate the “complexity” of curved surface shapes.K expresses the curve at arbitrary point p on a curved surface, and it is calculated as:κmax and κmin are principal curvatures  at p  at p . Informaormation , and it  κmaxκminwhere κmK at point v is approximately calculated, as . N. N19]. Nated, as :(4)K≅2π− ft at v . To get hape, as a:K’ = KDE and the state number V to define the states of K’. Subsequently, quantize K’ , as:(ii) Set the parameters for deviation E, V} = {30, 3} and {E, V} = {15, 15} are shown in s2 as same as points on a plane. Whereas, K’. Note that, methods to set parameters, which enables calculating entropy without bias, is proposed  and V is an odd number, as shown in E with high R2 becomes larger as V gets larger. HG when R2 is the largest value . This figure shows the R2 is 0.85 and the correspondence between HG and the sensory evaluation values. However, there are shapes whose values of HG differs while their sensory evaluation values are close, such as shapes 8 and 14 (8 and 14 .b. Result of Sample Shapes BE and V) and R2 of sample shapes B. R2 is higher when E is between  and V is an odd number, as shown in E with high R2 becomes larger as V gets larger. HG when R2 is the largest value . The R2 is 0.83 and the correspondence between HG and the sensory evaluation values is confirmed, as shown in HG is reversed, such as shapes 9 and 11  Shapes whose values of HG are higher in shape 8, while sensory evaluation values are close. Shape 8 has a concavity where values of K’ are low and this makes the value of HG lower than shape 14, which has no concavities, as shown in HG.(ii) Setting of ParametersE, V} = {200, 11}, which presents the highest value of R2. First, the setting of E is discussed, as follows. In shape 11, points at sharp convexities are allotted to s11 while points at concavities are allotted to s1. In shape 13, points in gradual convexities are allotted to s9 or s8, while points in gradual concavities are allotted to s4 or s3. In shape 6, points located out of convexities are allotted to s6. Therefore, it seems that various convexities and concavities of sample shapes A are allotted to different states that are based on their sharpness by setting parameters at {E, V} = {200, 11}.V is discussed, as follows. K’ when V = 3 or 4 in shape 6 of sample shapes A. When V = 3, the points on planes  are allotted to the same state, while points on corners are allotted to other states and the value of HG is 0.071. When V = 4, the points on planes are allotted to two states because of minute unevenness and the value of HG is 0.520. Therefore, it is appropriate to set V at an odd number to prevent overestimating minute unevenness.Next, setting of E with high R2 becomes larger as V gets larger is discussed. For example, the value of R2 becomes high not only when {E, V} = {200, 11} but also when {E, V} = {300, 15} or {300, 17}, as shown in K’ = 22, 67, 111, 166, and 200 when {E, V} = {200, 11}. On the other hand, boundaries are at K’ = 23, 69, 115, 162, and 208 when {E, V} = {300, 15} and K’ = 20, 60, 100, 140, and 180 when {E, V} = {300, 17}. Therefore, the tendency occurred because boundaries of K’ are similar among those settings of parameters. Further, cross-validation is carried out in Afterwards, the tendency that the value of b. Discussion about Sample Shapes BHG is reversed(i) Shapes whose relationship between the sensory evaluation values and HG were higher in shape 9 while the sensory evaluation values were higher in shape 11. Shape 9 has a sharp edge and points neighboring the edge are allotted to various states, as shown in HG rises. It seems that this is because the sampling points are not distributed along the edge and the values of K’ at the points are varied. Therefore, this can be solved by distributing sampling points along the edge during sampling. (ii) Setting ParametersE, V} = {50, 19}, which presents the highest value of R2. In shape 8, points at sharp convexities are allotted between s19 and s17, while points at sharp concavities are allotted to s1. In shape 5, the points at gradual convexities are allotted between s15 and s12, while points at gradual concavities are allotted to s9. In shape 6, the points at plane surfaces are allotted to s10. This suggests that the convexities and concavities of sample shapes B are allotted to different states based on their sharpness by setting parameters at {E, V} = {50, 19}. Note that the reason why V becomes an odd number is to prevent overestimating minute unevenness, the same as sample shapes A.HG and total absolute Gaussian curvature I is discussed. HG rather than I. In these shapes, sensory evaluation values are 3.38 and 3 and the values of HG are 0.210 and 0.152, respectively. However, values of I are 1.46 and 1.54, which are not corresponding to the sensory evaluation values. It seems that this is because I evaluates the number of concavities as “complexity” and I cannot evaluate the size of concavities. On the other hand, values of HG differ because the values of K’ differ in concavities with different size. Therefore, it seems that HG overcomes a shortcoming of I, which is unable evaluate “complexity” between shapes having the same number of concavities. Note that the tendency that the value of E with high R2 becomes larger as V gets larger is because the boundaries of K’ are similar among those settings of parameters, as same as sample shapes A. Further, cross-validation is carried out in At last, the difference between I rather than HG. In these shapes, sensory evaluation values are 2.71 and 2.79 and values of I are 1.59 and 1.66, respectively. However, values of HG are 0.044 and 0.057, which do not correspond to sensory evaluation values. It seems that this is because states of sampling points on edges are allotted to different states. On an edge of shape 4  weight abw of controlling points distributed on the k × l grid . Subsequently, fu was set at 0.004, which was 1/10 of 0.04. In addition, the parameters of HG were set at {E, V} = {2, 3}. This is because, in the preliminary experiment, the relationship between the sensory evaluation values about “complexity” and the value of HG of shapes generated by random transformation of the initial shape is examined and the R2 of logarithmic approximation on the relationship is maximized when {E, V} = {2, 3}. In this research, the method was applied to generate the shapes of Coupe-typed automobiles. At first, the NURBS surface was generated as the initial shape based on controlling points . Note th2. Experimental ConditionsEvaluation method: Just the same as x axis: 0 rpm, y axis: 0 rpm, z axis: 10 rpm), so that the appearance of all concavities can be observed.Sample shapes displaying method: White generated shapes on a black background were simultaneously displayed on a 13.3-inch laptop computer . During Presentation method: The distance between the eyeball of a participant and the computer was set to 500 mm. 15 sample shapes on the display simultaneously enter the field of view.Participants: 40 participants (36 men and four women) that ranged in age from 16 to 61 years .1. ResultsHG. The R2 of logarithmic approximation on the relationship is 0.71, and the correspondence between HG and the sensory evaluation values is confirmed. However, there are shapes that deviate from the approximate curve, such as shape 10, 11, and 15.2. DiscussionK’ on shapes that deviate from the approximate curve. These shapes have concavities at rear section and sampling points at the concavities are allotted to s1, as shown in this figure. It seems that the sensory evaluation value of the shape become lower, since it is difficult to recognize concavities at rear section. This is because it is natural to focus on the front section of the shape of an automobile rather than rear section. To examine this, HG considering only the front section is calculated and the correspondence to the sensory evaluation value is verified (R2 of logarithmic approximation is 0.89. Therefore, in calculating HG, it seems that it is necessary to consider the position of states and this can be achieved by weighing states based on their position.verified . ConsequHG and the sensory evaluation values. Moreover, knowledge about the parameters of Gaussian curvature entropy was obtained.This research proposed Gaussian curvature entropy as a “complexity” index of curved surface shapes by expanding curvature entropy, which is a “complexity” index of curves. Subsequently, correspondence between the index and human perceived “complexity” was examined by an experiment while using samples shapes generated by extrusion and rotation. Consequently, the coefficient of determinations between the value of the index and the sensory evaluation values about “complexity” were 0.85 and 0.83, and we confirmed the correspondence between Using the proposed index, this research constructed a curved surface shape generation method. The method was applied to generate the shapes of Coupe-typed automobiles and correspondence between the index and human perceived “complexity” of generated shapes was examined. The result shows the coefficient of determination between the value of the index and the sensory evaluation values regarding “complexity” was 0.71, and the usability of the method was confirmed. The result also shows the difficulty to recognize concavities at rear section of the shapes, and this decreases the sensory evaluation values. This means that the human bias focusing on the front section rather than rear section suggested the need to define the weight based on the section of shapes.The future study should consider the construction of a method for defining the weight based on their position during the calculation of Gaussian curvature entropy and carry out the validation experiment of the curved surface design by designers.


Zika virus infection during pregnancy can cause congenital brain and eye abnormalities and is associated with neurodevelopmental abnormalities . Jurisdictions were aggregated by level of local transmission of Zika virus: 1) widespread local transmission of Zika virus (Puerto Rico and the U.S. Virgin Islands); 2) limited local transmission of Zika virus ; and 3) without local transmission of Zika virus.Prevalence estimates for birth defects per 1,000 live births were calculated by group for each quarter. A PR  was calculated for each quarter. PRs and CIs were calculated using Poisson regression. SAS  was used to conduct all analyses.During January 1, 2016–June 30, 2017, among 2,004,630 live births, 3,359 infants and fetuses with a birth defect potentially related to Zika virus infection were delivered to residents of the 22 jurisdictions, including 2,813 (83.7%) with brain abnormalities and/or microcephaly and 546 (16.3%) with eye abnormalities without mention of a brain abnormality  . During https://stacks.cdc.gov/view/cdc/84198).The prevalence of birth defects potentially related to Zika virus infection in widespread local transmission areas was significantly higher in three periods during July 2016–March 2017 compared with that during the reference period. Prevalence increased fourfold  during January–March 2017 , compared with that during the reference period  , reachinOverall, most cases  had no reported laboratory testing of maternal, placental, fetal, or infant specimens. Among the remaining 191 cases, laboratory evidence of confirmed or possible Zika virus infection was reported in at least one specimen for 73 (2.2%) cases, and 118 (3.5%) had negative Zika virus laboratory testing. In widespread local transmission areas, laboratory testing at any time in at least one specimen was reported for 105 of 129 (81.4%) cases; among the 105 cases with laboratory testing, 50 (47.6%) had laboratory evidence of confirmed or possible Zika virus infection.The peak occurrence of birth defects potentially related to Zika virus infection in areas with widespread local transmission occurred in February 2017, 6 months after the reported peak of the Zika virus outbreak in these areas in August 2016 (Compared with the previous report (The findings in this report are subject to at least four limitations. First, results might not be generalizable beyond the included jurisdictions because jurisdictions might differ in population demographics and case-finding methodology. Second, heightened awareness can result in better identification of affected infants. For example, there might have been more extensive implementation of recommendations for eye exams in widespread local transmission areas. Third, categorization of areas with limited local transmission included regions of Florida and Texas that were larger than the actual areas of local transmission, which might mask any increase in Zika-related birth defects in smaller geographic areas where transmission occurred. Finally, the majority of cases did not have Zika virus testing reported. In widespread local transmission areas, approximately three quarters of cases had at least one sample tested, although the relatively high prevalence of negative results could reflect that timing might not have been optimal for detection of Zika virus in many cases. However, nearly half of those tested had laboratory evidence of Zika virus infection.During the Zika virus outbreak, population-based birth defects surveillance programs were adapted to monitor birth defects potentially related to Zika virus infection during pregnancy. Use of population-based birth defects surveillance programs and the U.S. Zika Pregnancy and Infant Registry provide an example of a complementary approach in ascertaining both exposures and outcomes to better monitor new and emerging threats during pregnancy and impact on infants (In states and territories with documented local Zika virus transmission, the prevalence of birth defects potentially related to Zika virus infection during pregnancy increased 21% during the second half of 2016 compared with that in the first half.In U.S. territories with widespread local Zika virus transmission, the prevalence of birth defects potentially related to Zika virus infection increased fourfold during January–March 2017 compared with January–March 2016.During the Zika virus outbreak, birth defects surveillance programs adapted to rapidly identify Zika-related birth defects regardless of laboratory evidence. These data provide more complete information on all infants affected and allow planning for care.


Marble waste particulates were found to considerably increase the tensile, as well as flexural, strength of the sustainable composites, which varied from 22.06 to 30.65 MPa and 43.27 to 58.11 MPa, respectively, for the molding temperature of 160 °C. The impact strength of the sustainable composites was found to surge with the increment in filler concentration, and the maximum impact strength was recorded as 1.66 kJ/m2 with 20% particulates reinforcement at a molding temperature of 200 °C. The thermal conductivity of the particulates-reinforced sustainable composites was as low as 0.23 W m−1K−1 at a 200 °C molding temperature with 20% and 40% filler concentrations, and the maximum thermal conductivity was 0.48 W m−1K−1 at a 160 °C molding temperature with 80% filler concentration. Our findings have shown a technically feasible option for manufacturing a lightweight composite with better mechanical and thermal properties using marble waste particulates as a potential civil infrastructural material.Marble waste is generated by marble processing units in large quantities and dumped onto open land areas. This creates environmental problems by contaminating soil, water, and air with adverse health effects on all the living organisms. In this work, we report on understanding the use of calcium-rich marble waste particulates (MPs) as economic reinforcement in recyclable polypropylene (PP) to prepare sustainable composites via the injection molding method. The process was optimized to make lightweight and high-strength thermally insulated sustainable composites. Physicochemical, mineralogical, and microscopic characterization of the processed marble waste particulates were carried out in detail. Composite samples were subsequently prepared via the injection molding technique with different filler concentrations  on weight fraction at temperatures of 160, 180, and 200 °C. Detailed analysis of the mechanical and thermal properties of the fabricated composites was carried out. The composites showed a density varying from 0.96 to 1.27 g/cm Marble processing units generate huge quantities of waste during different maneuvers such as cutting and polishing processes. It is produced in a slurry form, carried by the drainage system, and is discarded in open land areas, which, on drying, creates severe environmental issues. This accumulated marble waste contaminates water and air, and has harmful effects on plants, animals, and human health ,2,3,4. AThe effective use of marble waste particulates in the development of innovative advanced materials will serve the purpose of protecting the earth and becoming clean and green. Sustainable polymer composites may be one of the finest solicitations to use waste marble for replacing the traditional fillers, clay, and other conventional materials ,12,13. UResearchers have attempted to use marble waste particulates in paver blocks , soil stThe appropriate combination of marble waste particulates with silica and carbon black was used as a potential reinforcing material in the manufacturing of hybrid composites using natural rubber as the matrix material . Marble As the generation of industrial wastes such as marble waste causes economic and environmental problems, the potential use of marble waste particulates in developing hybrid polymer composites can be realized. In this work, we have made efforts to effectively reuse marble waste particulates (MPs) to develop a lightweight and high-strength sustainable hybrid polymer composite in a polypropylene (PP) system for different construction applications. To achieve this objective, physicochemical characterization of the received marble waste particulates was carried out by determining moisture content, particle size, density, energy-dispersive X-ray spectroscopy (EDS), specific gravity, pH, and electrical conductivity. Morphological and mineralogical analysis of marble waste particulates was carried out by state-of-the-art SEM and X-ray techniques. MP-PP composite samples were prepared by the injection molding technique with different filler concentrations of 0%, 20%, 40%, 60%, and 80% at injection molding temperatures of 160, 180, and 200 °C.The marble waste particulates (MPs) used was collected from a marble processing unit in Udaipur, Rajasthan, India. Polypropylene (PP) purchased from Three Star Plastics, Grade No. AM 650 N was used as a binder in the injection molding machine. Marble waste particulates were heated at 110 °C for 24 h, cooled in desiccators, and then sieved through a BS 150 (105 µm) sieve. Granulated PP was used as a polymer matrix with sulfur vulcanization as the catalyst. Sulfur vulcanization resultedincross-linking thermoplastic, which is non-toxic, safe to handle, and rapidly curing at about 150 °C with no tendency to scorch. Moreover, it is non-volatile, prevents loss during mixing, and acts as effective reinforcing filler.3)2(dolomite), CaCO3 and SiO2(quartz) are the mineral phases confirmed by XRD analysis in marble waste used in this study. After processing the marble waste particulates (heating at 110 °C for 24 h and then cooled in desiccators and sieved through a BS 150/105 µm sieve), it was mixed with granular polypropylene (PP), and sustainable MP-PP composites were prepared using an injection molding machine at 160, 180, and 200 °C. After the experimental investigation carried out in the present study, it was found that the prepared sustainable composites were green materials manufactured by marble waste and were low density, energy-saving, high-strength, recyclable, thermally insulative, and water-resistant (For manufacturing hybrid polymer composites using marble waste particulates (MPs), these particulates were mixed with the polypropylene (PP), and composites were prepared using an injection molding machine  at 160, 180, and 200 °C. Marble waste particulates as filler were used at concentrations of 0, 20, 40, 60, and 80 wt. % in the polypropylene (PP) system. A schematic diagram for the fabrication of the hybrid composite using marble waste particulates with polypropylene is shown in esistant .The moisture content of the obtained marble waste particulate sample was determined as per IS: 2720 (Part II)—1973 (Reaffirmed 1997) by the oven drying method. At 110 ± 5 °C, the average water content of the received sample was determined . The speW1 and W2 are the wet weight and conditioned weight of the sample.ASTM D 792-13 was used to determine the density of MP-PP composites (Equation (1)). The water absorption of composites was analyzed using ASTM D 570-98 (Reapproved 2018). Distilled water was used to submerge each sample separately at 25 ± 2 °C for the specified time interval of 24 h to determine the water absorption of the composite as per the previous protocol (Equation (2)) ,42,43.). Samples were analyzed using a WDW-50Universal testing machine , using a load cell of 50 kN and a gauge length of 50 mm with a 5 mm/min cross-head speed ,44. Flex3, and 2.58 ± 0.03, respectively. The mean particle size of waste particulates was found to be 32.62 ± 0.61 µm. By the particle size distribution curve, it was observed that in the marble waste particulates used, 90% of the total volume had a diameter smaller than 70.96 µm (D90), 50% of the total volume had a diameter smaller than 23.66 µm (D50), and 10% of the total volume had a diameter smaller than 6.98 µm (D10). Particle size analysis depicts that the particle size was in the range of 6.98 to 70.96 µm   . The pH The chemical composition of marble waste particulates studied by EDS analysis is shown in 90), 50% of the total volume had a smaller diameter than 23.66 µm (D50), and 10% of the total volume had a smaller diameter than 6.98 µm (D10). The mean particle size of the marble waste particulate sample was 32.62 ± 0.61 µm. Undersize (volume %) shows a broad distribution extending from a minimum of 1.318 to a maximum of 174.616 µm.The obtained marble waste particulate sample’s particle size distribution was determined by a dynamic laser scattering analyzer, and 3)2) phase (JCPDS card no. 84-1208), quartz (SiO2) phase (JCPDS card no. 86-1629), and calcite (CaCO3) mineral phases (JCPDS card no. 05-0586). It is also clear that the marble waste particulate sample has a crystal structure and the concentration of the dolomite phase is higher than other minerals.The mineralogical study of marble waste particulates was carried out using X-ray diffraction analysis in the 2θ range of 20°–70°. The morphology and size of marble waste particulates were investigated by the digital microscopy technique. Digital micrographs of the marble waste particulate sample are shown in Density, as well as water absorption, studies were carried out for pure PP and marble waste particulates-based polypropylene composites. 3 at 160 °C and 0.85 g/cm3 at 180 and 200 °C. MP-PP composites prepared at 160 °C with 20, 40, 60, and 80 wt. % filler concentrations showed densities of 0.98, 1.09, 1.18, and 1.27 g/cm3, respectively. MP-PP composites prepared at 180 °C with 20, 40, 60 and 80 wt. % filler concentrations showed densities of 0.97, 1.09, 1.18, and 1.19 g/cm3, respectively. MP-PP composites prepared at 200 °C with 20, 40, 60, and 80 wt. % filler concentrations showed densities of 0.96, 1.07, 1.18, and 1.26 g/cm3, respectively.The density of pure polypropylene (PP) was found to be 0.87 g/cm3 for all the injection molding temperatures of 160, 180, and 200 °C. The density of the MP-PP composite was a minimum of 0.96 g/cm3 at a 200 °C molding temperature with a 20 wt. % filler concentration and a maximum of 1.27 g/cm3 at a 160 °C molding temperature with a 80 wt. % filler concentration.It was observed that the density of pure PP slightly decreased with an increase in molding temperature and the density of MP-PP composites increased with the increment in the concentration of the filler ,46. For To determine the water absorption of the fabricated composite samples as per Equation (2), three observations were taken, and their mean value is presented in Water absorption of 0.008% was observed for pure PP samples fabricated at 160 and 180 °C injection molding temperatures. At a 200 °C molding temperature, pure PP samples showed 0.007% water absorption. MP-PP composites prepared at 160 °C with 20, 40, 60, and 80 wt. % filler concentrations showed water absorptions of 0.015, 0.019, 0.018, and 0.017%, respectively. MP-PP composites prepared at 180 °C with 20, 40, 60, and 80 wt. % filler concentrations showed water absorptions of 0.015, 0.026, 0.018, and 0.006%, respectively. MP-PP composites prepared at 200 °C injection molding temperature with 20, 40, 60, and 80 wt. % filler concentrations showed water absorptions of 0.013, 0.034, 0.024, and 0.022%, respectively. The results indicated that with an increase in filler concentration up to 40%, the water absorption increases, and beyond 40% filler concentration, this decreases in the case of all the injection molding temperatures of 160, 180, and 200 °C ,46. The The mechanical properties of injection-molded pristine PP and marble waste particulate polymer composites with filler concentrations of 20, 40, 60, and 80 wt. % in a PP system at injection molding temperatures of 160, 180, and 200 °C were determined . The tenThe tensile strength of pristine PP and marble waste particulate (MP)-filled polymer composite samples b fabricaThe flexural strength of pristine PP and marble waste particulate (MP)-filled polymer composites d fabrica2, respectively. The impact strengths of 0, 20, 40, 60, and 80 wt. % filler-concentration polymer composite samples fabricated at an injection molding temperature of 180 °C were found to be 1.32, 1.40, 1.42, 0.79 and 0.77 kJ/m2, respectively. The impact strengths of 0, 20, 40, 60, and 80 wt. % filler-concentration polymer composite samples fabricated at an injection molding temperature of 200 °C were found to be 1.34, 1.66, 1.58, 0.98, and 0.86 kJ/m2, respectively. It is worth noting that the impact properties of the fabricated sustainable composites rise with the increment in filler concentration up to 60, 40, and 20 wt. % for samples prepared at 160, 180, and 200 °C, respectively, and beyond these percentages, it decreases (2 at a 200 °C molding temperature with a 20 wt. % filler concentration (The impact strength of pristine PP and marble waste particulate (MP)-filled polymer composite samples f fabricaecreases e [14,18,ntration .Fractured surfaces of the tensile/flexural/impact test samples are displayed in The thermal conductivity of pure PP and MP-PP composites has been measured as a function of filler concentration at room temperature. To determine the thermal conductivity of the fabricated composite samples as per Equation (9), three observations were taken, and their mean value is presented in −1K−1 for samples fabricated at 160 °C and0.24 W m−1K−1 for samples fabricated at 180 and 200 °C. MP-PP composites prepared at 160 °C with 20, 40, 60, and 80 wt. % filler concentrations showed thermal conductivities of 0.27, 0.32, 0.42, and 0.48 W m−1K−1, respectively. MP-PP composites prepared at 180 °C with 20, 40, 60, and 80 wt. % filler concentrations showed thermal conductivities of 0.26, 0.29, 0.37, and0.39 W m−1K−1, respectively. MP-PP composites prepared at a 200 °C injection molding temperature with 20 and 40 wt. % filler concentrations showed a thermal conductivity of 0.23 W m−1K−1, and with 60 and 80 wt. % filler concentrations showed thermal conductivities of 0.29 and 0.33 W m−1K−1, respectively. The results indicated that the thermal conductivity increases with the increment in concentration of marble waste filler because the filler provides the path for heat transfer except in case of the samples prepared at an injection molding temperature of 200 °C, where the value of thermal conductivity slightly decreases for 20% and 40% filler concentrations [−1K−1 at a 200 °C molding temperature with 20 and 40 wt. % filler concentrations. The maximum value of the thermal conductivity of the MP-PP composite has been observed as 0.48 W m−1K−1 at a 160 °C molding temperature with an80 wt. % filler concentration.The thermal conductivity of pure PP was found to be 0.23 W mtrations ,47. It i3 and a very low water absorption of 0.006–0.034%. Marble waste particulates polypropylene sustainable composites have been developed with a tensile and flexural strength varying from 22.06 to 30.65 MPa and 43.27 to 58.11 MPa, respectively, for the molding temperature of 160 °C. The impact strength of the sustainable composites has been found to increase with filler concentration and the maximum impact strength was recorded as 1.66 kJ/m2 with a 20% filler concentration at a molding temperature of 200 °C. The thermal conductivity of particulates-reinforced sustainable composites was as low as 0.23 W m−1K−1 at a 200 °C molding temperature with 20% and 40% filler concentrations, and the maximum thermal conductivity was 0.48 W m−1K−1 at a 160 °C molding temperature with a80% filler concentration. The present study has shown that the lightweight, waterproof composite with better mechanical and thermal properties using marble waste particulates can be manufactured as a potential civil infrastructural material.The results of the physicochemical analysis of marble waste particulates have indicated that this can be potential filler for the development of hybrid polymer composites for multiple applications. It is worth noting from the present study that the decrease in mechanical strength with the increase in filler content may be due to the agglomeration phenomenon of filler particles forming large clusters of filler in the polymer matrix and poor interfacial bonding between the filler and polymer. As a result, the surface contact area between the particles and the matrix decreases ,49. To o3, and 2.58 ± 0.03, respectively. The mean particle size of marble waste particulates was 32.62 ± 0.61 µm and the pH was 6.58 ± 0.26. An electrical conductivity of 0.065 ± 0.02dS/m was recorded and the elemental analysis showed the presence of carbon, calcium, and magnesium as major constituents followed by a small concentration of aluminum, silicon, manganese, sodium, iron, potassium, and titanium. The XRD results confirmed the presence of quartz (SiO2), dolomite (CaMg(CO3)2), and calcite (CaCO3) mineral phases.It is evident from this study that the moisture content, bulk density, and specific gravity of the marble waste particulates were found to be 1.80 ± 0.09%, 1.39 ± 0.01 g/cm3) at 200 °C molding temperature with 20 % filler concentration, increases in density were recorded with an increase in filler concentration, and maximum density (1.27 g/cm3) was recorded at 160 °C molding temperature with 80% filler concentration. The water absorption of the MP-PP composite found to be very low (0.006%) at a 180 °C molding temperature with 80 %filler concentration. The tensile and flexural strength of MP-PP composite was little higher , at 160 °C molding temperature with 20% filler concentration. The impact properties of fabricated composites rose with the increment in filler concentration up to 60, 40, and 20 %for samples prepared at 160, 180, and 200 °C, respectively. The maximum impact strength of the MP-PP composite was 1.66 ± 0.05 kJ/m2 at 200 °C molding temperature with 20 %filler concentration. The thermal conductivity value of the MP-PP composite was observed as 0.23 W m−1K−1 at 200 °C molding temperature with 20 and 40% filler concentrations, and the maximum thermal conductivity was recorded as 0.48 W m−1K−1 at 160 °C molding temperature with 80% filler concentration.The MP-PP composite showed a low density composite (0.96 g/cmThe results reveal that marble waste particulates can be a potential filler and reinforcement material with polypropylene system for the development of lightweight, waterproof, high-strength, and low-thermally conductive hybrid polymer composites for multiple applications.


Millet is a dangerous weed in crop fields. A lack of seed dormancy helps it to spread easily and be present in maize, wheat, and other crop fields. Our previous report revealed the possibility that millet can also play a role as a virus reservoir. In that study, we focused on visual symptoms and detected the presence of several viruses in millet using serological methods, which can only detect the presence of the investigated pathogen. In this current work, we used small RNA high-throughput sequencing as an unbiased virus diagnostic method to uncover presenting viruses in randomly sampled millet grown as a volunteer weed in two maize fields, showing stunting, chlorosis, and striped leaves. Our results confirmed the widespread presence of wheat streak mosaic virus at both locations. Moreover, barley yellow striate mosaic virus and barley virus G, neither of which had been previously described in Hungary, were also identified. As these viruses can cause severe diseases in wheat and other cereals, their presence in a weed implies a potential infection risk. Our study indicates that the presence of millet in fields requires special control to prevent the emergence of new viral diseases in crop fields. Panicum miliaceum subsp. miliaceum, is cultivated, but nearly 50 other species are listed as weeds. The results of a recent national weed survey in Hungary revealed that P. miliaceum was the seventh-most prevalent weed, representing 1.44% of the weeds present in the investigated fields. Moreover, the presence of other Panicum species  was also detected [Millet is used to feed animals and is a popular alternative cereal. However, as an outcome of recent intensification of crop production, it has also become a dangerous weed, controlled only with difficulty. Herbicides, based on chloraminothiazine, efficiently repress the growth of dicotyledonous weeds in crop fields and leave monocotyledonous millet without a competitor ,2,3,4. Cdetected ,7,8,9,10Millet can be infected by several viruses. To investigate its role as a potential virus reservoir in our previous work, we surveyed the natural viral infection of millet using DAS ELISA and found that 19 of the 45 samples were virus infected with wheat streak mosaic virus (WSMV), wheat dwarf virus (WDV), barley stripe mosaic virus (BSMV), barley yellow dwarf virus (BYDV), and brome streak mosaic virus (BStMV), either alone or in combination .While serological testing can only reveal the presence of the investigated pathogen, new metagenomics-based studies can reveal the entire virome of the samples and describe the presence of all of the presenting viruses, even if their presence could have not been anticipated . High-thWhile crops are widely surveyed for the presence of infecting viruses, knowledge about the viromes of natural flora remains limited. A pioneering geometagenomics study investigating the impact of agriculture on the distribution and prevalence of viruses reached the conclusion that virus prevalence is greater in cultivated areas and that agriculturally important viruses also infect uncultivated plant species . The impTritimovirus genus of the Potyviridae family, is a devastating pathogen in wheat-growing regions globally and was diagnosed in Hungary in the 1980s [Aceria tosichella) vector [WSMV, the type member of the ) vector . The hos) vector ,22. It i) vector .Laodelphax striatellus) [Barley yellow striate mosaic virus (BYSMV) is a cytorhabdovirus, having a negative single-stranded RNA genome and first being detected in its planthopper vector (atellus) . Althougatellus) . In addiatellus) , BYSMVs atellus) , Iran [2atellus) ,28, Syriatellus) , and Chiatellus) —and in Matellus) . Howeveratellus) , and thuBarley virus G (BVG), a recently described polerovirus, was first found in South Korea in barley . SubsequIn this work, we further investigated the possible virus reservoir role of millet grown as a volunteer weed at two locations using sRNA HTS as an unbiased diagnostic method to be able to detect infection with any viruses. The results confirmed the prevalent presence of WSMV. Moreover, we diagnosed BYSMV and BVG for the first time in Hungary.Panicum miliaceum plants at two locations in western Hungary near lake Balaton  coding region of the WSMV genome revealed that most of the sampled individuals were infected with WSMV. Seven and nine individuals at BA and US were found to be infected, respectively .According to the sequenced siRNAs, several variants were present, which explains why we cloned and sequenced 13 clones from each location, resulting in eight and nine slightly different variants (named according to their place of origin): WSMV_HUBA1-HUBA8 and WSMV_HUUS1-HUUS9.  Sequence comparison showed that they all cluster with strains in Clade B .The most characteristic feature of the WSMV clades is the lack of a single codon within the CP cistron, resulting in a lack of the Gly2761 codon within the CP coding region of the polyprotein in Clade B. This characteristic feature is present in all of the sequenced variants as in the previously sequenced Hungarian strain  and in aOur knowledge about millet as a host of WSMV is scarce, as millet has not been included in detailed host range studies either of crops or perennial grasses ,41. HoweAlthough it has not previously been described in Hungary, according to the sRNA HTS results, both sampled places were infected with BYSMV. BYSMV-derived sRNAs were mainly 21 nt long and equally distributed along the whole genome of this cytorhabdovirus in both libraries . In contThe RT-PCR validation test of the sampled plants, amplifying the CP coding region of the virus, revealed the presence of BYSMV in three (at BA) and one (at US) plant(s) . The se.This means that slightly different variants were present, which suggests they can be distributed not only at one location but also at a close distance. Sequence information about the BYSMV CP coding region is only available from the reference genome and not from the variants of different geographical origins. Hence, their phylogenetical analyses were unable to help us hypothesize about the original source of infection.For more detailed analysis of the phylogenetical origin of the new Hungarian BYSMV strains, deducting sequences from the sRNA HTS would be straightforward. Unfortunately, however, although the sRNA reads are evenly distributed in the genome, the coverage, which was just above 60%, does not allow us to directly undertake this comparison. This feature; low coverage of the viral genome, could be characteristic for the cytorhabdoviruses because, during the comparison of sRNA HTS and RNAseq, Pecman and colleagues found that the presented cytorhabdovirus could be detected only by RNAseq due to the low number of short reads . SequencAt US, sRNA HTS showed the presence of BVG, a recently described polerovirus . BGV-oriRT-PCR validation with BVG-specific primers, amplifying the CP coding region, revealed that only one plant  was infected with the virus .Phylogenetic analysis of the BVG_HUUS showed that it clusters distantly from the strains reported from Australia and the Proso millet isolates from South Korea. Its closest homologue is the BVG strain from switchgrass .To the best of our knowledge, with the exception of the report from the Netherlands, BVG has not been reported in Europe. It is therefore surprising that we found this virus in millet weed; it is difficult to explain its presence or speculate about its origin. Plant 18, which was infected with the virus, did not show any specific symptom . AlthougPanicum miliaceum plants were sampled randomly, representing different symptoms. We collected leaves together with root and millet seeds if it was possible . We purified sRNA fractions with the size separation on polyacrylamide gel-electrophoresis and followed our in-house-modified protocol  based onFor virus diagnosis, the bioinformatics analysis of the HTS results was carried out using a CLC Genomic Workbench . After the trimming and quality control of the reads, a QC report was prepared by applying embedded protocols in the CLC Genomic Workbench. Longer contigs were built from the non-redundant reads using an assembler of CLC  see . If at lPooled RNA extracts representing each sampled plant, and the plantation pool that was used for the sRNA HTS, were used as templates for cDNA synthesis by RevertAid First Strand cDNA Synthesis Kit  with random primers according to the manufacturer’s instructions. Viruses were diagnosed by RT-PCR performed with Q5 Hot Start High-Fidelity DNA Polymerase  (the primers used to amplify viral parts together with the used annealing temperatures and cycling parameters are provided in For phylogenetic analysis of viral sequences, we used MEGA7 . EvolutiOur pilot study shows that sRNA HTS can reveal new insights of investigated plants and lead to the description of viruses, BYSMV and BGV in our case, even if they have not been previously described or are not anticipated at the surveyed location. In addition, we found a high infection rate with WSMV supporting the hypothesis of Schubert and colleagues that viruses can establish themselves in seed production areas and survive on perennial grasses growing at field boundaries . This hiOur results suggest that, in addition to its direct competitive effect, millet could serve as a virus reservoir and might play a role in the spread of some cereal viruses. Thus, the control of millet weed, as well as the prevention of its overwintering and volunteer growth, is of paramount importance. However, to determine the importance and frequency of this type of virus distribution, further studies are needed. Insect vectors play a pivotal role in virus spread. Analysis of their virome, which is a new area of investigation , can add


Schistosoma haematobium (commonly known as bilharzia) continues to be largely ignored by national and global health policy-makers. International attention for large-scale action against the disease focuses on whether it is a risk factor for the transmission of human immunodeficiency virus (HIV). Yet female genital schistosomiasis itself is linked to pain, bleeding and sub- or infertility, leading to social stigma, and is a common issue for women in schistosomiasis-endemic areas in sub-Saharan Africa. The disease should therefore be recognized as another component of a comprehensive health and human rights agenda for women and girls in Africa, alongside HIV and cervical cancer. Each of these three diseases has a targeted and proven preventive intervention: antiretroviral therapy and pre-exposure prophylaxis for HIV; human papilloma virus vaccine for cervical cancer; and praziquantel treatment for female genital schistosomiasis. We discuss how female genital schistosomiasis control can be integrated with HIV and cervical cancer care. Such a programme will be part of a broader framework of sexual and reproductive health and rights, women’s empowerment and social justice in Africa. Integrated approaches that join up multiple public health programmes have the potential to expand or create opportunities to reach more girls and women throughout their life course. We outline a pragmatic operational research agenda that has the potential to optimize joint implementation of a package of measures responding to the specific needs of girls and women.Female genital schistosomiasis as a result of chronic infection with  Schistosoma haematobium (commonly known as bilharzia) has been described in the medical literature since the 1940s.,,Gynaecological schistosomiasis as a result of chronic infection with Female genital schistosomiasis has not yet been reviewed or assessed as a specific entity, nor as part of the burden of schistosomiasis, by the Global Burden of Diseases Study.S. haematobium deoxyribonucleic acid in vaginal lavage has a good positive predictive value for genital lesions, especially during the earlier stages of female genital schistosomiasis .A standard diagnosis of female genital schistosomiasis requires colposcopy and histopathology and is often only conducted in well-equipped facilities that are not widely available in endemic areas. Community-based assessment of the disease burden is therefore challenging. Studies investigating the disease have been carried out in 18 countries in Africa, with a cumulative total of only 10 332 women examined . In Nigeria the prevalence of female genital schistosomiasis in four schistosomiasis-endemic communities was 27.4% based on self-reported symptoms (87 of 317 women tested) and 70.0% as confirmed by gynaecological examination (14 women in a subsample of 20).A recent study in Zambia aimed to validate community-based diagnosis of female genital schistosomiasis by genital self-swabs. The prevalence of active schistosomiasis was 5.5% (33 out of 603 women) as detected by urine microscopy and 15.1% (91 out of 601 women) by circulating anodic antigen testing.,–Female genital schistosomiasis has been suspected as a risk factor for the transmission of human immunodeficiency virus (HIV) for more than 25 years, based on pathophysiological, immunological and epidemiological data.S. haematobium infection and female genital schistosomiasis.,While the link between HIV and human papilloma virus (HPV) has been the subject of a systematic review and meta-analysis,At the local level, within communities and the health workforce, there are gaps in knowledge and understanding about female genital schistosomiasis prevention, treatment and control. Health-care providers can easily confuse the symptoms of the disease with those of sexually transmitted infections or cervical cancer. Incorrect diagnosis and management can have profound psychosocial implications for pre-sexually active girls. The effect of the disease on fertility and pregnancy may have serious consequences for a woman’s relationships with her family and community, often resulting in marginalization, stigma, isolation and the threat of gender-based violence.Female genital schistosomiasis is a serious gynaecological condition linked to pain, bleeding and severe social stigma in Africa. The consequences of the disease merit an urgent response in the broader sexual and reproductive health and rights agenda of girls and women in sub-Saharan Africa.–Women and girls face multiple and intersecting health, social, gender and economic challenges, especially in the poorest and most marginalized and fragile parts of our world. Many, if not most of these women and girls in sub-Saharan Africa carry a triple burden of vulnerability to HIV, HPV/cervical cancer and female genital schistosomiasis. There is mounting evidence for a three-way interaction between these diseases and that controlling one may decrease the risk of unwanted outcomes for the two others.A high-level meeting was held to break the deadlock to large-scale implementation of female genital schistosomiasis interventions and to facilitate an integrated approach with HIV and cervical cancer prevention and care. Over a hundred international experts gathered on the side-lines of the 11th European Congress on Tropical Medicine and International Health in September 2019.The operational research needs that were highlighted during the meeting included: (i) community-level burden assessment of female genital schistosomiasis; (ii) strategies for ensuring regular praziquantel treatment of women and girls outside of school-based deworming programmes; (iii) clinical treatment protocols for existing female genital schistosomiasis; (iv) strategies to improve awareness of female genital schistosomiasis and community engagement with the issue; and (v) impact evaluation and economic analysis of integrated strategies versus separate, parallel approaches.The Bilharzia and HIV study is currently evaluating genital self-swabbing as a potential low-cost approach to community-based diagnosis for female genital schistosomiasis in Zambia.Chronic female genital schistosomiasis lesions can be prevented by regular treatment with praziquantel started at an early age, such as through primary school-based deworming programmes,While its preventive effect is well documented, it is less clear whether praziquantel treatment at the standard dose improves established genital disease. In the Bilharzia and HIV Study in Zambia of women aged 18–31 years, the frequency of six female genital schistosomiasis-related symptoms all declined by 5 to 10 percentage points after a single dose . This finding is consistent with the partial resolution of lesions observed in Malawi,Box 1How can we mobilize and engage communities, health services and their workforce so that female genital schistosomiasis is understood and acted upon appropriately and stigma is avoided? What are the enabling factors and barriers for the integration of female genital schistosomiasis, HIV and HPV/cervical cancer in sexual and reproductive health and rights programmes at the community level? How can we rapidly build the capacity of the skilled health workforce, front-line health workers and school teachers to become agents of change in favour of fair and comprehensive sexual and reproductive health and rights programmes? How can we increase community awareness of female genital schistosomiasis prevention, treatment and control and meaningfully engage sexual and reproductive health and rights and HIV activists? How can community activists become advocates within their communities and agents of demand for inclusive, non-stigmatizing, integrated and comprehensive services? ,How can community awareness and demand creation influence policy changes to address the issues of safe water and behavioural change to prevent schistosomiasis?HIV: human immunodeficiency virus; HPV: human papilloma virus.All these are operational research questions that biomedical and social science researchers will need to jointly answer. Many lessons can be drawn from the experience of HIV with regards to mobilizing and engaging communities, amplifying the call for integrated services and creating demand for service and commodities to address the specific needs of women and girls when and where they need them.More studies to strengthen the evidence base for a cause-and-effect relationship among female genital schistosomiasis, HIV and HPV/cervical cancer would clearly be useful. Randomized controlled trial evidence would be ideal, but proven and well-established interventions are available for each of these diseases and cannot be withheld in a trial setting. Although research on neglected tropical diseases remains relatively underfunded, the lack of robust evidence should not be a deterrent to moving forward. Close monitoring and evaluation of the impact of a comprehensive response on all three diseases could potentially be nested into ongoing population-based impact assessment and evaluation efforts. The data should provide the answer to the question of whether implementation of the packages of care provides enhanced benefits, should be scaled up and offers opportunities for economies of scale.The physical and psychosocial impact of female genital schistosomiasis, along with the potential links across the three priority genital diseases, position female genital schistosomiasis as a pillar in a comprehensive sexual and reproductive health and rights agenda for women and girls in Africa.Although female genital schistosomiasis is preventable by regular treatment, too often praziquantel is not available within the primary health-care system outside of school-based, large-scale treatment.Established female genital schistosomiasis can be diagnosed during screening for cervical cancer, such as colposcopy and visual inspection of the cervix with acetic acid. The World Health Organization (WHO) has produced a visual aid to facilitate recognition of female genital schistosomiasis by clinical health-care professionals.Preventive interventions for HIV, cervical cancer and female genital schistosomiasis can all be successfully delivered through existing programmes. An integrated programme would strengthen health systems and make a compelling case for a strategic change towards comprehensive sexual and reproductive health and rights services in affected regions . A pragmSeveral relevant global initiatives are already in place: the sustainable development goals, including universal health coverage;


We hypothesised that intercarpal K-wire fixation of adjacent carpal bones would reduce torque and lever force within a fractured scaphoid bone.In eight cadaver wrists, a scaphoid osteotomy was stabilised using a locking nail, which also functioned as a sensor to measure isometric torque and lever forces between the fragments. The wrist was moved through 80% of full range of motion (ROM) to generate torque and force within the scaphoid. Testing was performed with and without loading of the wrist and K-wire stabilisation of the adjacent carpal bones.P = 0.0485). Other than wrist extension, torque direction was not predictable.Average torque and lever force values were 49.6 ± 25.1 Nmm and 3.5 ± 0.9 N during extension and 41 ± 26.7 Nmm and 8.1 ± 2.8 N during flexion. Torque and lever force did not depend on scaphoid size, individual wrist ROM, or deviations of the sensor versus the anatomic axis. K-wire fixation did not produce significant changes in average torque and lever force values except with wrist radial abduction (In unstable scaphoid fractures, we suggest securing rotational stability with selected implants for functional postoperative care. Wrist ROM within 20% extension and radial abduction to 50% flexion limit torque and lever force exacerbation between scaphoid fragments. The scaphoid is the most frequently fractured bone in the wrist and presents clinical challenges that include inadequate diagnosis as well as healing. Among these, vascular supply has been viewed as the main reason for problems in treating scaphoid fractures and non-unions . Recent The wrists of eight anonymous cadaver forearms  were passively moved in four directions  following scaphoid osteotomy and reconnection with an implanted isometric sensor. The resulting change in torque relative to wrist positioning was measured. The methodology, in particular surgical technique, sensor structure, force distribution, and forearm fixation, was developed in a pilot study and recently published .The setup in this investigation consisted of a forearm and a mobile hand fixation system with an integrated goniometer to record angles of wrist movement in two orthogonal planes. In addition, the system allowed for preloading the wrist Table  using a Guided movement of the wrist was performed with and without joint loading in all four directions. Angle of the wrist movement sequences and torque within the scaphoid were continuously recorded. The setup allowed guidance of uniaxial wrist movements and prevented deviations in other directions. Each individual movement cycle was repeated three times and controlled manually by an operator until resistance indicated the end of the wrist’s ROM. To exclude interobserver variability, a single operator carried out all measurements for maximal ROM. As a way to avoid excessive tension towards the end of the ROM, repetitive movement cycles were evaluated only up to 80% of the entire ROM. Figure x and y directions were not recorded for this investigation. A standardised osteotomy of the scaphoid was performed on the palmar side in a lateral direction within the middle third of the scaphoid. The capsulotomy required for the osteotomy was closed with Ethibond® suture .To measure the biomechanical interaction between the two fracture segments of the osteotomised scaphoid, we developed a sensor that also served as a rigidly binding osteosynthesis implant Figs.  and 3. TTo further stabilise the scaphoid fragments, we introduced two K-wires from the distal scaphoid fragment into the capitate and from the capitate into the lunate Fig. . AfterwaFollowing sensor explantation, we documented the position of the drilling canals within the scaphoid fragments used high-resolution peripheral quantitative computed tomography . With this tool, we analysed the position of the sensor and the osteotomy within the scaphoid bone. After these measurements, the position and quality of the implanted sensor and condition of the ligaments were examined during dissection. The cadaver wrists were examined for arthritis and an intact ligamentous structure after the experiment was conducted. Loosening of the sensor was ruled out by dissection following testing. Hand and scaphoid sizes varied considerably, so we used water displacement to measure the scaphoid volume. Because of the small dimension, incompletely transected scaphoids and transection of a sensor cable forced us to abandon six preparations, and we had to process 14 cadaver hands to obtain a sample size of 8.Absolute torque and force were measured at 20%, 50%, and 80% of total ROM in each wrist movement direction  and for each combination of the two experimental factors K-wire arthrodesis (with K-wire vs without K-wire) and wrist loading (loaded vs unloaded). All measurements were repeated four times. Prior to the statistical analysis, these four repeated measurements were averaged to obtain a single torque or force value under each experimental condition. The means and standard deviations of torque and force were calculated for each experimental condition.F-test. To account for performing 48 hypothesis tests in total, Bonferroni–Holm-adjusted P values are reported in addition to unadjusted P values. We also calculated P values for the null hypothesis of no interaction between the factors K-wire and wrist loading. We viewed these tests as exploratory and did not include them in the multiple testing adjustment.To further analyse the effect of K-wire arthrodesis and wrist loading on torque and force, we fit separate linear mixed models for force and torque values obtained under a given combination of total ROM and wrist movement direction. The models included the factors K-wire arthrodesis, wrist loading, and their interaction as explanatory variables. Scaphoid identity was included as a random factor to account for the dependency between observations of the same scaphoid under different experimental conditions. From these models, we calculated the mean differences between the two levels of each factor (averaging across the levels of the other factor). The null hypothesis of a true mean difference of zero was tested using an All experiments were carried out in accordance with relevant guidelines and regulations. This study was approved by the local institutional ethics board .P values from 0.0569 to 1.000) in any model but did so in torque through 80% radial duction   of 7.1  Nmm in the mean torque at 20% ROM when the wrist was loaded. For all other ROM values, the corresponding increase in torque and force was not significant . The highest average torque values within the scaphoid were measured at 80% of flexion in the unloaded (33.9 ± 28.9 Nmm) and loaded states (41.0 ± 26.7 Nmm). The highest average values of force within the scaphoid were measured at 80% of flexion in the unloaded (5.3 ± 3.6 N) and loaded states (8.1 ± 2.8 N).Flexion of the wrist was guided to 54.5 ± 10.7° and to 54.9 ± 10.04° during loading. K-wire arthrodesis limited flexion to 42.5 ± 10.2° without loading and to 43.8 ± 12.1° with it. Wrist flexion from 20 to 80% ROM caused torque levels between the scaphoid fragments from 3.9 ± 3.4 to 33.9 ± 28.9 Nmm and force levels from 0.5 ± 0.3 to 5.3 ± 3.6 N. We observed a significant increase  of 9 Nmm  in the mean torque at 20% ROM, a significant increase  of 20.6  Nmm in the mean torque at 50% ROM, a significant increase  of 1.5  N in the mean force at 20% ROM, and a significant increase  of 2.1  N in the mean force at 50% ROM. For all other values of ROM, the corresponding increase in torque and force by loading was not significant . Highest average values of torque within the scaphoid were measured at 80% of extension in the unloaded (22 ± 17.5 Nmm) and loaded states (49.6 ± 25.1 Nmm). Highest average values for force within the scaphoid were measured at 80% of extension in the unloaded (1.9 ± 1 N) and loaded states (3.5 ± 0.9 N).Wrist extension was guided to 77.6 ± 16.04° and to 71.46 ± 13.4° during loading. K-wire arthrodesis limited extension to 58.75 ± 18.8° without and to 52.4 ± 19° with wrist loading. Wrist extension without loading from 20 to 80% of ROM caused torque levels between the scaphoid fragments from 5.2 ± 3.6 to 22 ± 18.9 Nmm and force levels from 0.4 ± 0.2 to 1.9 ± 1 N. When the wrist was loaded, we observed a significant increase  of 7.5 Nmm in the mean torque at 20% ROM by loading of the wrist. For all other values of ROM, the corresponding increase in torque was not significant.Radial abduction of the wrist was guided to 23.3 ± 5.18° and to 24.9 ± 4.23° during loading. K-wire arthrodesis limited radial abduction to 17.52 ± 6.55° without and to 17.8 ± 6.52° with loading of the wrist. Radial abduction of the wrist without loading from 20 to 80% of ROM caused torque levels between the scaphoid fragments from 2.2 ± 0.8 to 52.2 ± 36.3 Nmm and force levels from 0.3 ± 0.2 to 2.9 ± 1.7 N. We observed a significant increase  of 0.9  N in the mean force at 20% ROM, a significant increase  of 2.7  N in the mean force at 50% ROM, and a significant increase  of 3.5  N in the mean force at 80% ROM. The highest average values of torque within the scaphoid were measured at 80% of radial abduction in the unloaded state (52.2 ± 36.3 Nmm) and in the loaded state (52.4 ± 45.5 Nmm). The highest average values of force within the scaphoid were measured at 80% of wrist radial abduction in the unloaded (2.9 ± 1.7 N) and loaded states (5.6 ± 2 N).When the wrist was loaded, we observed a significant increase  of 16.6  Nmm in the mean torque at 50% ROM and a significant increase  of 21.8  Nmm in the mean torque at 80% ROM by loading of the wrist. We observed a significant increase  of 1.4  N in the mean force at 20% ROM, a significant increase  of 2.8  N in the mean force at 50% ROM, and a significant increase  of 3  N in the mean force at 80% ROM by loading of the wrist. For all other ROM values, the corresponding increase in torque was not significant. The highest average values of torque within the scaphoid were measured at 80% of ulnar abduction in the unloaded (4.7 ± 6.1 Nmm) and loaded states (23.6 ± 16 Nmm). Highest average values of force within the scaphoid were measured at 80% of wrist ulnar abduction in the unloaded (0.6 ± 0.6 N) and loaded states (4.7 ± 2.2 N). In summary, our results suggest that tendon loading significantly affects torque and force within the scaphoid during all moving directions, depending on ROM.Ulnar abduction of the wrist was guided to 31.37 ± 1.26° and to 31.5 ± 1.59° during loading. K-wire arthrodesis limited ulnar abduction to 27.77 ± 5.5° without and to 28.47 ± 7.9° with loading of the wrist. We observed a significant increase  values. Other than wrist extension for torque and wrist flexion for lever force, the directions of the developed torque and lever force, respectively, were not predictable.Here, we used an intraosseous sensor to examine isometric torque through the longitudinal axis of the scaphoid. Slade et al. suspected that the scaphoid is subjected to rotatory forces throughout the wrist ROM and confirmed their hypothesis that torque is generated within the scaphoid during wrist motion . End-ranOur results cannot be directly compared to those of Fortis et al., who used rosette strain gauges to investigate how wrist motion influences the magnitude of the specific strain and compression on the intact scaphoid . Other aCentrally placed scaphoid screws can improve construct stiffness compared with eccentrically placed scaphoid screws . Indeed,A uniform direction of force within a sector of 90° radiopalmar direction of coronal scaphoid plane could be determined only in wrist flexion. Conversely, a uniform direction of torque could be determined only in extension of the wrist; thus, we can draw no conclusions regarding the rotational direction in which a scaphoid osteosynthesis screw provides more stability. In our opinion, the rigidity of a scaphoid osteosynthesis using a single bone screw will prevent more lever forces along the longitudinal axis than rotational torques generated from bypassing the lever forces.To minimise the force of the investigator, torque was examined only at 20%, 50%, and 80% of the respective ROM. Because high torque values > 0.1 Nm were measured in 80% of full ROM in flexion, radial duction, and extension in some of the individuals, even higher torques can be expected and must be withstood by single-screw osteosyntheses in vivo because patients will perform movements to the full end-ROM. In our investigation, from 20% extension to 50% flexion, we measured low torque values below 25% of average torque at 80% of ROM. Based on these results, we cannot recommend a preferred movement direction. Rather, as clinical postoperative guidelines after screw osteosyntheses, we suggest a defined limited ROM from 20% extension and radial abduction to 50% flexion. Important parameters affecting torque in the scaphoid involve individual wrist factors. However, we could not determine that the varying sizes of the scaphoid and the longitudinal axis affected the fluctuations in torque and lever force between the specimens during wrist motion. In addition, K-wire intercarpal stabilisation did not reduce torque magnitude between the two fragments. Our current data do not allow us to provide further details to clarify the cause of the varying torque and force magnitudes between the specimens.As studied on long bones, evidence is limited on the biomechanics involved in fracture healing of the scaphoid bone , 22. DurIn addition, with our experimental setup, we could not identify scaphoid size as an influencing factor. Other anatomical variations such as ligamentous laxity, scaphoid shape, and intercarpal arrangement cannot be investigated with our experimental approach but are likely to cause variations in measured forces.The design and usage of the sensor also involved several limitations. The in vitro application differed from the calibrating procedure because only one end was fixed in a chuck for lever force measurement. For torque calibration, one end was fixed in every direction, and the other end was stabilised against the lever force but kept free in rotation to avoid combined torque and lever force. In vitro, both ends were fixed to ligament-stabilised but mobile bone fragments. The result was a combination of flexibility in rotation and bending along the sensor axis. Discrepancy in necessary tendon-pulling forces to achieve comparable wrist motion did not allow for force-controlled wrist motion in our experiments.Because of a lack of sensor surface area, we used half-bridges for force measurement. For the sensor construction, Wheatstone full-bridges would have been best not only for torque but also for lever force measurement to eliminate simultaneous upsetting force and thermal loading orthogonal to the sensor axis.In addition to interrupted circulation, prolonged healing after single-screw osteosynthesis might arise because of an existing instability occurring during wrist loading and motion , 25. WriOur results add to understanding of the biomechanics of the scaphoid and indicate the need for greater fixation rigidity in fracture cases and non-unions. These findings serve as a basis for additional experiments to examine the level of torque and lever force that a screw-fragment construct should be able to withstand.Additional file 1: Supplementary Figure 1: Force within the scaphoid at 20%, 50%, and 80% of wrist ROM  in the unloaded and loaded states with and without K-wire stabilisation.Additional file 2: Supplementary Figure 2: Lever force direction between the scaphoid fragments during wrist motion at 80% of ROM of any wrist movement direction.Additional file 3: Supplementary Figure 3: Lever force direction between the scaphoid fragments during wrist motion at 80% of ROM of any wrist movement. In this condition, the wrist was partially blocked by K-wires.