IntroductionBacillus anthracis is the causative agent for the lethal disease anthrax, primarily affecting animals and humans in close contact with an infected host. The pathogenicity of B. anthracis is attributed to the secreted exotoxins and their outer capsule. The host cell-binding exotoxin component “protective antigen” (PA) is reported to be a potent vaccine candidate. The aim of our study is to produce several PA constructs and analyze their vaccine potential.MethodsWe have designed the various subunit, PA-based recombinant proteins, i.e., full-length Protective antigen (PA-FL), C-terminal 63 kDa fragment (PA63), Protective antigen domain 1-domain 4 chimeras (PA-D1-4) and protective antigen domain 4 (PA-D4) and analyzed their vaccine potential with different human-compatible adjuvants in the mouse model. We have optimized the process and successfully expressed our recombinant antigens as soluble proteins, except full-length PA. All the recombinant antigen formulations with three different adjuvants i.e., Addavax, Alhydrogel, and Montanide ISA 720, were immunized in different mouse groups. The vaccine efficacy of the formulations was analyzed by mouse serum antigen-specific antibody titer, toxin neutralization assay, and survival analysis of mouse groups challenged with a lethal dose of B. anthracis virulent spores.ResultsWe have demonstrated that the PA-FL addavax and PA63 addavax formulations were most effective in protecting spore-challenged mice and serum from the mice immunized with PAFL addavax, PA-FL alhydrogel, PA63 addavax, and PA63 alhydrogel formulations were equivalently efficient in neutralizing the anthrax lethal toxin. The higher levels of serum Th1, Th2, and Th17 cytokines in PA-FL addavax immunized mice correspond to the enhanced protection provided by the formulation in challenged mice.DiscussionWe have demonstrated that the PA-FL addavax and PA63 addavax formulations exhibit equivalent efficiency as vaccine formulation both in a mouse model of anthrax and mammalian cell lines. However, PA63 is a smaller antigen than PA-FL and more importantly, PA63 is expressed as a soluble protein in E. coli, which imparts a translational advantage to PA63-based formulation. Thus, the outcome of our study has significant implications for the development of protective antigen-based vaccine formulations for human use against the lethal disease anthrax.

In recent years, study of particulate matter become an important public health concern. Small particals PM2.5, which have diameter less than 2.5 micro meter impacts on lung diseases and respiratory system of human. A number of various computational techniques are there to estimate the concentration of these particles present in the atmosphere. In this paper, Random Forest Regressor (RFR) is proposed to estimate the concentration of PM2.5 particles. Model is trained on 11 different features i.e. annual average temperature (T), maximum temperature (MT), minimum temperature (mT), rain precipitation (RP), average wind speed (WS), total rainy days (RD), total snowy days (SD), total stormy days (StD), total foggy days (FD), total tornado days (TD), total haily days (HD). Data is collected through web scrapping for the Bangalore city, India from year 2013 to 2020. Model performance obtained was R2=0.9732, MAE=3.87μg/m3, and RMSE=2.84μg/m3. Simulated result showed higher accuracy over other existing techniques.

The presence of particulate matter, in the atmospheric environment, affects the health of living creatures as well as the ecosystem. Estimation of particulate matter has become one of the most challenging study for researchers. There are numerous computer techniques for the estimation of these particles. In this study, a multi kernel support vector machine (M-SVM) approach is introduced for the categorisation of particulate matter captured as digital images. Images from the archive of many outdoor scene (AMOS) have been taken for implementation purpose. The model is trained to predict the level of particulate matter captured as a digital image. An experimental model with M-SVM classification predicts the particulate matter captured as image among three levels, i.e., whether an image has a normal level, critical level or highly critical level. Simulated results were found to analyse the particulate matter with 98% of accuracy, which ensures efficient recognition of our experimental method.

Over the last few years, the study of characteristics of particles present in the environment becomes an interesting research area for scientists. Simulation of physical and dynamic characteristics of particulate matter (PM) is a prominent area for researchers. For the implementation of particle image velocimetry (PIV) for complex particulate matter with overlapping boundaries, it is necessary to remove non-physical measurements. These non-physical measurements such as unsteady surface and inaccurate edge detection lead to the spurious velocity of particles. In this note, a Canny edge detector is employed to identify the edge of particles. For unsteady surfaces, a special process is followed as follows: (a) find the gradient magnitude in the particles image velocimetry frame from the Canny edge detected frame to optimize particle detection, (b) classify a large high-intensity area in view to extract the particles, (c) find out the rough surface area which contains these particles with their reflections, (d) finally eliminate these particle's reflections. Finally, after this, PIV is implemented on these extracted processed frames from the video dataset to measure the motion of the particles. In this paper, a Canny edge detector with particle image velocimetry (PIV) algorithm is proposed to simulate the dynamic behavior of particulate matter present in particles stock video footage (PSVF) dataset of particles. The proposed model is trained to estimate the motion of particles, and the result showed an accuracy of 92.97% for the particles stock video footage dataset over the other existing methods.

Plasmodium falciparum Cysteine-Rich Protective Antigen (CyRPA) is an essential, highly conserved merozoite antigen that forms an important multi-protein complex (RH5/Ripr/CyRPA) necessary for erythrocyte invasion. CyRPA is a promising blood-stage vaccine target that has been shown to elicit potent strain-transcending parasite neutralizing antibodies. Recently, we demonstrated that naturally acquired immune anti-CyRPA antibodies are invasion-inhibitory and therefore a correlate of protection against malaria. Here, we describe a process for the large-scale production of tag-free CyRPA vaccine in E. coli and demonstrate its parasite neutralizing efficacy with commonly used adjuvants. CyRPA was purified from inclusion bodies using a one-step purification method with high purity (>90%). Biochemical and biophysical characterization showed that the purified tag-free CyRPA interacted with RH5, readily detected by a conformation-specific CyRPA monoclonal antibody and recognized by sera from malaria infected individuals thus indicating that the recombinant antigen was correctly folded and retained its native conformation. Tag-free CyRPA formulated with Freund's adjuvant elicited highly potent parasite neutralizing antibodies achieving inhibition of >90% across diverse parasite strains. Importantly, we identified tag-free CyRPA/Alhydrogel formulation as most effective in inducing a highly immunogenic antibody response that exhibited efficacious, cross-strain in vitro parasite neutralization achieving ~80% at 10 mg/ml. Further, CyRPA/Alhydrogel vaccine induced anti-parasite cytokine response in mice. In summary, our study provides a simple, scalable, cost-effective process for the production of tag-free CyRPA that in combination with human-compatible adjuvant induces efficacious humoral and cell-mediated immune response.

Plasmodium falciparum cysteine-rich protective antigen (CyRPA) is a conserved component of an essential erythrocyte invasion complex (RH5/Ripr/CyRPA) and a target of potent cross-strain parasite-neutralizing antibodies. While naturally acquired human RH5 antibodies have been functionally characterized, there are no similar reports on CyRPA.

Customer’s opinion holds a lot of importance for the business to succeed. The reviews and the ratings obtained are helpful to analyse the products and improve the marketing strategy for the products and gain profits. The conventional methods used for the product reviews include the star ratings system and textual feedback of the product. The latter is more descriptive and reliable and better in terms of providing genuine feedback of the product. This paper discusses and implements the sentiment analysis using Naïve Bayes model to obtain the sentiment rating of the product, later, sarcasm detection using Sequential model is integrated to obtain and improve the overall sentiment rating of the product. An aspect-based sentiment analysis (ABSA) is also integrated to analyse the product on various aspects. This methodology will help sellers and customers to save time and understand how a particular product is performing in the market. The results obtained will help them gain a clear understanding that if the product is a success or not.

Serodiagnosis of surra, caused by Trypanosoma evansi, is still based on native antigens purified from bloodstream form of T. evansi grown in rodents. In order to investigate prospective diagnostic possibilities as an alternative for native antigens, we cloned, expressed 26 kDa calflagin protein containing 218 amino acids from T. evansi (Indian Strain) in Escherichia coli. The potential of recombinant calflagin (rCLF) protein as diagnostic antigen was evaluated in immunoblot and indirect ELISA using experimentally infected equine serum samples from 0 to 84 days post infection. The antibodies against T. evansi were detected with rCLF antigen in serum samples of experimentally infected equines as early as 10 days and 14 days post infection, using immunoblot and ELISA respectively. No cross-reactivity was observed with rCLF antigen in ELISA with different serum samples of equines positive for Equine herpesvirus 1, Burkholderia mallei, and Theileria equi infections. Several immunoreactive regions ranging from 10 to 28 kDa were detected using distinct T. evansi isolates (pony, cattle, donkey and camel origin) indicating presence of multiple calflagin family members in a single trypanosome. Indirect immunofluorescence antibody test with anti-CLF rabbit hyperimmune serum showed localisation of native immunogenic protein near attachment of flagellum. The rCLF protein was found to be a potential diagnostic candidate for distinguishing T. evansi positive and negative equine serum sample, suggesting that it could be used for serological surveys in animals for surra. In addition, it could be used with other potential diagnostic candidates to improve the diagnostic efficiency.

Introduction: Vibration is a known aeromedical stressor in rotary-winged aircraft and is known to affect visual acuity (VA). In aerospace operational conditions, contrast sensitivity (CS) takes the upper hand over VA for optimal visual performance. Examination of the effects of low frequencies whole body vibration of short duration on CS was the desired objective of the study. Material and Methods: Thirty healthy volunteers were exposed to low frequency whole body vibration using Multi-axial Vibration Simulator. Vibration frequency along the Z-axis varied from 4 to 20 Hz over a period of 30 min. CS was recorded in no vibration, under vibration and 30 min following exposure to vibration using CSV-1000 equipment at spatial frequencies of 3, 6, 12, and 18 cycles per degree (cpd). Results: CS, after an initial degradation, gradually improved with the increase in frequencies of vibration for the lower spatial frequencies (3, 6 cpd). The increasing trend was also noticed for higher spatial frequencies (12, 18 cpd) till 16 Hz; thereafter, a significant dip (P = 0.048) was observed at 20 Hz of vibration. No significant difference was observed following 30 min of post exposure. Conclusion: In contrast to the popular belief, an improvement in CS with increasing frequencies of vibration could be concluded from the results of the study. However, the sudden decrement in CS at higher CPD at higher frequencies of vibration could adversely affect visual performance of an aircrew.

Thermal Imaging refers to capturing the infrared radiation of the images and creating images out of the infrared radiation. This infrared radiation can be captured by special purpose infrared cameras. These cameras do not have the need to capture the images in ambient light. The images can be captured in darkness and can be used to detect objects and living beings. Object detection techniques earlier used for normal images using deep learning can be extended to thermal images. The paper discusses the various Convolution Neural Networks(CNN) models available to detect the objects. We then look at the YOLOv3 and Spatial Pyramid Pooling(SPP) approach to detect objects in thermal images. YOLOv3 approach to detection is revolutionary as it uses a single CNN to get the bounding boxes and class probabilities for the image. The approach discussed here contains YOLOv3 and SPP layer. YOLOv3 contains a deep convolution neural network, the convolution neural network, we then add a Spatial Pyramid Pooling(SPP) layer on top of the CNN. The SPP layer removes the need for cropping the image in the fully connected layer. This gives a speedup of over 100 times. The precision and accuracy goals of over 80 percent are met with the above approach for the FLIR dataset.

Coinfection with Plasmodium falciparum and helminths may impact the immune response to these parasites because they induce different immune profiles. We studied the effects of coinfections on the antibody profile in a cohort of 715 Mozambican children and adults using the Luminex technology with a panel of 16 antigens from P. falciparum and 11 antigens from helminths (Ascaris lumbricoides, hookworm, Trichuris trichiura, Strongyloides stercoralis, and Schistosoma spp.) and measured antigen-specific IgG and total IgE responses. We compared the antibody profile between groups defined by P. falciparum and helminth previous exposure (based on serology) and/or current infection (determined by microscopy and/or qPCR). In multivariable regression models adjusted by demographic, socioeconomic, water, and sanitation variables, individuals exposed/infected with P. falciparum and helminths had significantly higher total IgE and antigen-specific IgG levels, magnitude (sum of all levels) and breadth of response to both types of parasites compared to individuals exposed/infected with only one type of parasite (P ≤ 0.05). There was a positive association between exposure/infection with P. falciparum and exposure/infection with helminths or the number of helminth species, and vice versa (P ≤ 0.001). In addition, children coexposed/coinfected tended (P = 0.062) to have higher P. falciparum parasitemia than those single exposed/infected. Our results suggest that an increase in the antibody responses in coexposed/coinfected individuals may reflect higher exposure and be due to a more permissive immune environment to infection in the host. IMPORTANCE Coinfection with Plasmodium falciparum and helminths may impact the immune response to these parasites because they induce different immune profiles. We compared the antibody profile between groups of Mozambican individuals defined by P. falciparum and helminth previous exposure and/or current infection. Our results show a significant increase in antibody responses in individuals coexposed/coinfected with P. falciparum and helminths in comparison with individuals exposed/infected with only one of these parasites, and suggest that this increase is due to a more permissive immune environment to infection in the host. Importantly, this study takes previous exposure into account, which is particularly relevant in endemic areas where continuous infections imprint and shape the immune system. Deciphering the implications of coinfections deserves attention because accounting for the real interactions that occur in nature could improve the design of integrated disease control strategies.

Background: The evaluation of immune responses to RTS,S/AS01 has traditionally focused on immunoglobulin (Ig) G antibodies that are only moderately associated with protection. The role of other antibody isotypes that could also contribute to vaccine efficacy remains unclear. Here we investigated whether RTS,S/AS01(E) elicits antigen-specific serum IgA antibodies to the vaccine and other malaria antigens, and we explored their association with protection. Methods: Ninety-five children (age 5-17 months old at first vaccination) from the RTS,S/AS01(E) phase 3 clinical trial who received 3 doses of RTS,S/AS01(E) or a comparator vaccine were selected for IgA quantification 1 month post primary immunization. Two sites with different malaria transmission intensities (MTI) and clinical malaria cases and controls, were included. Measurements of IgA against different constructs of the circumsporozoite protein (CSP) vaccine antigen and 16 vaccine-unrelated Plasmodium falciparum antigens were performed using a quantitative suspension array assay. Results: RTS,S vaccination induced a 1.2 to 2-fold increase in levels of serum/plasma IgA antibodies to all CSP constructs, which was not observed upon immunization with a comparator vaccine. The IgA response against 13 out of 16 vaccine-unrelated P. falciparum antigens also increased after vaccination, and levels were higher in recipients of RTS,S than in comparators. IgA levels to malaria antigens before vaccination were more elevated in the high MTI than the low MTI site. No statistically significant association of IgA with protection was found in exploratory analyses. Conclusions: RTS,S/AS01(E) induces IgA responses in peripheral blood against CSP vaccine antigens and other P. falciparum vaccine-unrelated antigens, similar to what we previously showed for IgG responses. Collectively, data warrant further investigation of the potential contribution of vaccine-induced IgA responses to efficacy and any possible interplay, either synergistic or antagonistic, with protective IgG, as identifying mediators of protection by RTS,S/AS01(E) immunization is necessary for the design of improved second-generation vaccines. (C) 2020 Elsevier Ltd. All rights reserved.

Plasmodium falciparum, the parasite responsible for severe malaria, develops within erythrocytes. Merozoite invasion and subsequent egress of intraerythrocytic parasites are essential for this erythrocytic cycle, parasite survival and pathogenesis. In the present study, we report the essential role of a novel protein, P. falciparum Merozoite Surface Antigen 180 (PfMSA180), which is conserved across Plasmodium species and recently shown to be associated with the P. vivax merozoite surface. Here, we studied MSA180 expression, processing, localization and function in P. falciparum blood stages. Initially we examined its role in invasion, a process mediated by multiple ligand-receptor interactions and an attractive step for targeting with inhibitory antibodies through the development of a malaria vaccine. Using antibodies specific for different regions of PfMSA180, together with a parasite containing a conditional pfmsa180 -gene knockout generated using CRISPR/Cas9 and DiCre recombinase technology, we demonstrate that this protein is unlikely to play a crucial role in erythrocyte invasion. However, deletion of the pfmsa180 gene resulted in a severe egress defect, preventing schizont rupture and blocking the erythrocytic cycle. Our study highlights an essential role of PfMSA180 in parasite egress, which could be targeted through the development of a novel malaria intervention strategy.

The Industrial Internet of things or IIoT has gained recognition due to the advancement it has made in communication technology. Industrial IoT is an application of IoT that enables control of industries over the Internet using smart devices and sensors. The two main entity which ensures effectiveness in any field is monitoring and control. Keeping a view on this aspect, we have designed a low-cost, low-power Wi-Fi based industrial monitoring system that controls and monitors the remote manufacturing plants and industries using a web application. In this model an Arduino Mega which is the main micro-controller is connected with a Wi-Fi module for internet connectivity, a barometer sensor for temperature and pressure, a humidity sensor for sensing the humidity and a gas sensor which detects the smoke and harmful gases. These components are utilized to build a monitoring system. Apart from these components several other sensors are used to keep a check on the temperature, gas leakage, pressure, humidity, etc. in the work environment to ensure the workers safety. In case of any incident this monitoring system warns the workers by an alarm and sends information to the registered user via Blynk App. The chief purpose of this research is to sum up the significant role of IoT in monitoring industries.

The RTS,S/AS01E vaccine has shown consistent but partial vaccine efficacy in a pediatric phase 3 clinical trial using a 3-dose immunization schedule. A fourth-dose 18 months after the primary vaccination was shown to restore the waning efficacy. However, only total IgG against the immunodominant malaria vaccine epitope has been analyzed following the booster. To better characterize the magnitude, nature, and longevity of the immune response to the booster, we measured levels of total IgM, IgG, and IgG1-4 subclasses against three constructs of the circumsporozoite protein (CSP) and the hepatitis B surface antigen (HBsAg, also present in RTS,S) by quantitative suspension array technology in 50 subjects in the phase 3 trial in Manhiça, Mozambique. To explore the impact of vaccination on naturally acquired immune responses, we measured antibodies to P. falciparum antigens not included in RTS,S. We found increased IgG, IgG1, IgG3 and IgG4, but not IgG2 nor IgM, levels against vaccine antigens 1 month after the fourth dose. Overall, antibody responses to the booster dose were lower than the initial peak response to primary immunization and children had higher IgG and IgG1 levels than infants. Higher anti-Rh5 IgG and IgG1-4 levels were detected after the booster dose, suggesting that RTS,S partial protection could increase some blood stage antibody responses. Our work shows that the response to the RTS,S/AS01E booster dose is different from the primary vaccine immune response and highlights the dynamic changes in subclass antibody patterns upon the vaccine booster and with acquisition of adaptive immunity to malaria.

Artemisinin is a remarkable compound whose derivatives and combinations with multiple drugs have been utilized at the forefront of malaria treatment. However, the inherent issues of the parent compound such as poor bioavailability, short serum half-life, and high first-pass metabolism partially limit further applications of this drug. In this study, we enhanced the aqueous phase solubility of artemisinin by encapsulating it in two nanocarriers based on the polymer polycaprolactone (ART-PCL) and lipid-based Large Unilamellar Vesicles (ART-LIPO) respectively. Both nanoformulations exhibit in vitro parasite killing activity against Plasmodium falciparum with the ART-LIPO performing at comparable efficacy to the control drug solubilized in ethanol. These water-soluble formulations showed potent in vivo antimalarial activity as well in the mouse model of malaria at equivalent doses of the parent drug. Additionally, the artemisinin-PCL nanoformulation used in combination with either pyrimethamine or chloroquine increased the survival of the Plasmodium berghei infected mice for more than 34 days and effectively cured the mice of the infection. We highlight the potential for polymer and liposome-based nanocarriers in improving not only the aqueous phase solubility of artemisinin but also concomitantly retaining its therapeutic efficacy in vivo as well.

Bacillus anthracis poly-γ-D-glutamic acid (PGA) capsule is an essential virulent factor that helps the bacterial pathogen to escape host immunity. Like other encapsulated bacterial species, the B. anthracis capsule may also inhibit complement-mediated clearance and ensure bacterial survival in the host. Previous reports suggest that B. anthracis spore proteins inhibit complement activation. However, the mechanism through which the B. anthracis capsule imparts a survival advantage to the active bacteria has not been demonstrated till date. Thus, to evaluate the role of the PGA capsule in evading host immunity, we have undertaken the present head-to-head comparative study of the phagocytosis and complement activation of non-encapsulated and encapsulated B. anthracis strains. The encapsulated virulent strain exhibited resistance toward complement-dependent and complement-independent bacterial phagocytosis by human macrophages. The non-encapsulated Sterne strain was highly susceptible to phagocytosis by THP-1 macrophages, after incubation with normal human serum (NHS), heat-inactivated serum, and serum-free media, thus indicating that the capsule inhibited both complement-dependent and complement-independent opsonic phagocytosis. An increased binding of C3b and its subsequent activation to C3c and C3dg, which functionally act as potent opsonins, were observed with the non-encapsulated Sterne strain compared with the encapsulated strain. Other known mediators of complement fixation, IgG, C-reactive protein (CRP), and serum amyloid P component (SAP), also bound more prominently with the non-encapsulated Sterne strain. Studies with complement pathway-specific, component-deficient serum demonstrated that the classical pathway was primarily involved in mediating C3b binding on the non-encapsulated bacteria. Both strains equally bound the complement regulatory proteins C4BP and factor H. Importantly, we demonstrated that the negative charge of the PGA capsule was responsible for the differential binding of the complement proteins between the non-encapsulated and encapsulated strains. At lower pH closer to the isoelectric point of PGA, the neutralization of the negative charge was associated with an increased binding of C3b and IgG with the encapsulated B. anthracis strain. Overall, our data have demonstrated that the B. anthracis capsule inhibits complement fixation and opsonization resulting in reduced phagocytosis by macrophages, thus allowing the bacterial pathogen to evade host immunity.

Polymer and liposome-based nanocarriers not only improve the aqueous phase solubility of artemisinin but also helps to retain its therapeutic efficacyin vivoas well.