Search

Ixodes scapularis is a blood-feeding obligate ectoparasite responsible for transmitting the Lyme disease (LD) agent, Borrelia burgdorferi. During the feeding process, I. scapularis injects B. burgdorferi into the host along with its saliva, facilitating the transmission and colonization of the LD agent. Tick calreticulin (CRT) is one of the earliest tick saliva proteins identified and is currently utilized as a biomarker for tick bites. Our recent findings revealed elevated levels of CRT in the saliva proteome of B. burgdorferi-infected I. scapularis nymphs compared to uninfected ticks. Differential precipitation of proteins (DiffPOP) and LC-MS/MS analyses were used to identify the interactions between Ixs (I. scapularis) CRT and human plasma proteins and further explore its potential role in shielding B. burgdorferi from complement killing. We observed that although yeast-expressed recombinant (r) IxsCRT binds to the C1 complex (C1q, C1r, and C1s), the activator of complement via the classical cascade, it did not inhibit the deposition of the membrane attack complex (MAC) via the classical pathway. Intriguingly, rIxsCRT binds intermediate complement proteins (C3, C5, and C9) and reduces MAC deposition through the lectin pathway. Despite the inhibition of MAC deposition in the lectin pathway, rIxsCRT did not protect a serum-sensitive B. burgdorferi strain (B314/pBBE22Luc) from complement-induced killing. As B. burgdorferi establishes a local dermal infection before disseminating to secondary organs, it is noteworthy that rIxsCRT promotes the replication of B. burgdorferi in culture. We hypothesize that rIxsCRT may contribute to the transmission and/or host colonization of B. burgdorferi by acting as a decoy activator of complement and by fostering B. burgdorferi replication at the transmission site.

Lyme disease (LD) caused by Borrelia burgdorferi is among the most important human vector borne diseases for which there is no effective prevention method. Identification of tick saliva transmission factors of the LD agent is needed before the highly advocated tick antigen-based vaccine could be developed. We previously reported the highly conserved Ixodes scapularis (Ixs) tick saliva serpin (S) 17 (IxsS17) was highly secreted by B. burgdorferi infected nymphs. Here, we show that IxsS17 promote tick feeding and enhances B. burgdorferi colonization of the host. We show that IxsS17 is not part of a redundant system, and its functional domain reactive center loop (RCL) is 100% conserved in all tick species. Yeast expressed recombinant (r) IxsS17 inhibits effector proteases of inflammation, blood clotting, and complement innate immune systems. Interestingly, differential precipitation analysis revealed novel functional insights that IxsS17 interacts with both effector proteases and regulatory protease inhibitors. For instance, rIxsS17 interacted with blood clotting proteases, fXII, fX, fXII, plasmin, and plasma kallikrein alongside blood clotting regulatory serpins (antithrombin III and heparin cofactor II). Similarly, rIxsS17 interacted with both complement system serine proteases, C1s, C2, and factor I and the regulatory serpin, plasma protease C1 inhibitor. Consistently, we validated that rIxsS17 dose dependently blocked deposition of the complement membrane attack complex via the lectin complement pathway and protected complement sensitive B. burgdorferi from complement-mediated killing. Likewise, co-inoculating C3H/HeN mice with rIxsS17 and B. burgdorferi significantly enhanced colonization of mouse heart and skin organs in a reverse dose dependent manner. Taken together, our data suggests an important role for IxsS17 in tick feeding and B. burgdorferi colonization of the host.

Human hyaluronidase 1 (HYAL1) and PH20 play vital roles in degrading hyaluronic acids through the substrate-assisted double displacement mechanism. While HYAL1, a lysosomal enzyme, functions optimally under acidic conditions, PH20, a sperm surface hyaluronidase, displays a broader pH range, from acidic to neutral. Our objective was to extend HYAL1's pH range towards neutral pH by introducing repulsive charge-charge interactions involving the catalytic Glu131, increasing its pKa as the proton donor. Substituting individual acidic residues in the β3-loop (S77D), β3'-β3″ hairpin (T86D and P87E), and at Ala132 (A132D and A132E) enabled HYAL1 to demonstrate enzyme activity at pH 7, with the mutants S77D, P87E, and A132E showing the highest activity in the substrate gel assay. However, double and triple substitutions, including S77D/T86D/A132E as found in the PH20 configuration, did not result in enhanced activity compared to single substitutions. Conversely, PH20 mutants with non-acidic substitutions, such as D94S in the β3-loop and D103T in the β3'-β3″ hairpin, significantly reduced activity within the pH range of 4 to 7. However, the PH20 mutant E149A, reciprocally substituted compared to A132E in HYAL1, exhibited activity similar to PH20 wild-type (WT) at pH 7. In a turbidimetric assay, HYAL1 mutants with single acidic substitutions exhibited activity similar to that of PH20 WT at pH 7. These results suggest that substituting acidic residues near Glu131 results in HYAL1 activity at neutral pH through electrostatic repulsion. This study highlights the significance of charge-charge interactions in both HYAL1 and PH20 in regulating the pH-dependent activity of hyaluronidases.

Tick serine protease inhibitors (serpins) play crucial roles in tick feeding and pathogen transmission. We demonstrate that Ixodes scapularis (Ixs) nymph tick saliva serpin (S) 41 (IxsS41), secreted by Borrelia burgdorferi (Bb)-infected ticks at high abundance, is involved in regulating tick evasion of host innate immunity and promoting host colonization by Bb. Recombinant (r) proteins were expressed in Pichia pastoris, and substrate hydrolysis assays were used to determine. Ex vivo (complement and hemostasis function related) and in vivo (paw edema and effect on Bb colonization of C3H/HeN mice organs) assays were conducted to validate function. We demonstrate that rIxsS41 inhibits chymase and cathepsin G, pro-inflammatory proteases that are released by mast cells and neutrophils, the first immune cells at the tick feeding site. Importantly, stoichiometry of inhibition analysis revealed that 2.2 and 2.8 molecules of rIxsS41 are needed to 100% inhibit 1 molecule of chymase and cathepsin G, respectively, suggesting that findings here are likely events at the tick feeding site. Furthermore, chymase-mediated paw edema, induced by the mast cell degranulator, compound 48/80 (C48/80), was blocked by rIxsS41. Likewise, rIxsS41 reduced membrane attack complex (MAC) deposition via the alternative and lectin complement activation pathways and dose-dependently protected Bb from complement killing. Additionally, co-inoculating C3H/HeN mice with Bb together with rIxsS41 or with a mixture (rIxsS41 and C48/80). Findings in this study suggest that IxsS41 markedly contributes to tick feeding and host colonization by Bb. Therefore, we conclude that IxsS41 is a potential candidate for an anti-tick vaccine to prevent transmission of the Lyme disease agent.

Summary: The clinical (re)development of bacteriophage (phage) therapy to treat antibiotic-resistant infections faces the challenge of understanding the dynamics of phage-bacteria interactions in the in vivo context. Here, we develop a general strategy coupling in vitro and in vivo experiments with a mathematical model to characterize the interplay between phage and bacteria during pneumonia induced by a pathogenic strain of Escherichia coli. The model allows the estimation of several key parameters for phage therapeutic efficacy. In particular, it quantifies the impact of dose and route of phage administration as well as the synergism of phage and the innate immune response on bacterial clearance. Simulations predict a limited impact of the intrinsic phage characteristics in agreement with the current semi-empirical choices of phages for compassionate treatments. Model-based approaches will foster the deployment of future phage-therapy clinical trials.

Abstract Background Schistosoma japonicum, which inhabits the mesenteric vein of the mammalian hosts for about 20 to 30 years, is subjected to the oxidative stresses from the host defense mechanism during their intra-mammalian stages. To counteract this host immune attack, the parasite utilizes their antioxidant system for survival inside the host. Peroxiredoxins (Prxs), thiol-specific antioxidant proteins, play an essential role for protecting the parasite against oxidative stress by reducing hydrogen peroxide to water. Only three types of 2-Cys Prxs have been previously characterized in S. japonicum whereas a fourth Prx has been identified for Schistosoma mansoni as Prx-4. A sequence coding homologous to this gene in the S. japonicum database was identified, characterized and expressed as recombinant SjPrx-4 protein (rSjPrx-4). Furthermore, rSjPrx-4 was evaluated in this study for its diagnostic potentials in detecting S. japonicum infection in humans. Results The gene found in the parasite genome contained 2 active-site cysteines with conserved sequences in the predicted amino acid (AA) sequence and showed 75% identity with that of the previously characterized Prx (TPx-1) of S. japonicum. The gene was expressed in different stages of schistosome life-cycle with highest transcription level in the adult male. The gene was cloned into a plasmid vector and then transfected into Escherichia coli for expression of rSjPrx-4. Anti-rSjPrx-4 mouse sera recognized native SjPrx-4 in egg and adult worm lysate by western blotting. The result of a mixed function oxidation assay in which rSjPrx-4 prevented the nicking of DNA from hydroxyl radicals confirmed its antioxidant activity. Subsequently, immunolocalization analysis showed the localization of SjPrx-4 inside the egg, on the tegument and in the parenchyma of the adult worm. Enzyme-linked immunosorbent assay results showed that rSjPrx-4 has 83.3% sensitivity and 87.8% specificity. Its diagnostic potential was further evaluated in combination with recombinant SjTPx-1 protein, yielding an improved sensitivity and specificity of 90% and 92.7%, respectively. Conclusions These results suggest that SjPrx-4 plays a role as an antioxidant dealing with oxidative stresses of S. japonicum, and its diagnostic potential improved by coupling it with SjTPx-1 is a proof for developing a serological test with better diagnostic performance for human schistosomiasis.

Bovine babesiosis is a tick-transmitted disease caused by intraerythrocytic protozoan parasites of the genus Babesia. Its main causative agents in the Americas are Babesia bigemina and Babesia bovis, while Babesia ovata affects cattle in Asia. All Babesia species secrete proteins stored in organelles of the apical complex, which are involved in all steps of the invasion process of vertebrate host cells. Unlike other apicomplexans, which have dense granules, babesia parasites instead have large, round intracellular organelles called spherical bodies. Evidence suggests that proteins from these organelles are released during the process of invading red blood cells, where spherical body proteins (SBPs) play an important role in cytoskeleton reorganization. In this study, we characterized the gene that encodes SBP4 in B. bigemina. This gene is transcribed and expressed in the erythrocytic stages of B. bigemina. The sbp4 gene consists of 834 nucleotides without introns that encode a protein of 277 amino acids. In silico analysis predicted a signal peptide that is cleaved at residue 20, producing a 28.88-kDa protein. The presence of a signal peptide and the absence of transmembrane domains suggest that this protein is secreted. Importantly, when cattle were immunized with recombinant B. bigemina SBP4, antibodies identified B. bigemina and B. ovata merozoites according to confocal microscopy observations and were able to neutralize parasite multiplication in vitro for both species. Four peptides with predicted B-cell epitopes were identified to be conserved in 17 different isolates from six countries. Compared with the pre-immunization sera, antibodies against these conserved peptides reduced parasite invasion in vitro by 57%, 44%, 42%, and 38% for peptides 1, 2, 3, and 4, respectively (p < 0.05). Moreover, sera from cattle infected with B. bigemina cattle contained antibodies that recognized the individual peptides. All these results support the concept of spb4 as a new gene in B. bigemina that should be considered a candidate for a vaccine to control bovine babesiosis.

The present study aims to measure the number of cycles leading to fracture (NCF) of instruments in static and dynamic cyclic fatigue situations under body temperature in stainless steel double-curved canals. The framework was constructed to establish the movement of instruments occurring at a stable body temperature. A step motor, a holding system for an endodontic handpiece, created the movement in and out of the artificial canal of the file mounted on the handpiece. A total of 30 instruments of ProTaper Universal and ProTaper Next (Dentsply Sirona, Maillefer, Ballaigues, Switzerland) were divided into three groups of 10 per group. For group 1 (10 PTU F2), files were rotated in static cycles. For groups 2 (10 PTU F2) and 3 (10 PTN X2), files were rotated in dynamic cycles. Files were rotated using proprietary programs, and the times the files were rotated before fracture were recorded. Data were analyzed using survival probabilities and regression with life data. The ProTaper Next in the dynamic cycles had the largest NCF, and the ProTaper Universal in the static condition had the least. New modes of rotation, material, and design have affected the cyclic fatigue resistance of the instrument.

Thioredoxin (Trx), a small redox protein, exhibits thermal stability at high temperatures regardless of its origin, including psychrophiles. Trxs have a common structure consisting of the central β-sheet flanked by an aliphatic cluster on one side and an aromatic cluster on the other side. Although the roles of aromatic amino acids in the folding and stability of proteins have been studied extensively, the contributions of aromatic residues to the stability and function of Trx, particularly Trxs from cold-adapted organisms, have not been fully elucidated. This study examined the roles of aromatic amino acids in the aromatic cluster of a Trx from the psychrophilic Arctic bacterium Sphingomonas sp. PAMC 26621 (SpTrx). The aromatic cluster of SpTrx was comprised of W11, F26, F69, and F80, in which F26 at the β2 terminus was buried inside. The substitution of tyrosine for F26 changed the SpTrx conformation substantially compared to that of F69 and F80. Further biochemical and spectroscopic investigations on F26 showed that the F26Y, F26W, and F26A mutants resulted in structural instability of SpTrx in both urea- and temperature-induced unfolding and lower insulin reduction activities. The Trx reductase (SpTR) showed lower catalytic efficiencies against F26 mutants compared to the wild-type SpTrx. These results suggest that buried F26 is essential for maintaining the active-site conformation of SpTrx as an oxidoreductase and its structural stability for interactions with SpTR at colder temperatures.

This paper presents a survey dataset on factors influencing farmers' intention to produce organic farming in Hanoi, Vietnam. The survey was designed based on the theoretical integration model of theory of planned behavior (TPB) and norm activation model (NAM) including 7 factors, 33 items inherited from previous studies to collect information of the respondents and 5 other items used to find out the respondent's characteristic include: gender, age, educational qualification, farming experience and farming annual income. Questionnaires were sent directly to farmers at their home or farm in October 2019. 318 valid questionnaires were collected for the study of intentions for organic farming. The dataset was obtained as a reference source for later studies on organic farming development and organic farming production intent/behavior promotion.

In Mongolia, horses play important roles, not only in livestock production, but also in terms of culture, tradition, and Mongolian beliefs. Although the presence of non-tsetse-transmitted horse trypanosomoses, which are caused by infections with Trypanosoma evansi (surra) and T. equiperdum (dourine), has been reported in the country, whether there is a nationwide epidemic of these infectious diseases is unknown. In the present study, a nationwide surveillance of horse trypanosomoses was performed. The sample sizes for each province, the whole country, and male and female horses were, respectively, 96, 2,400, and 316 and 306. In total, 3,641 samples of horse sera were collected by simple random sampling. The rTeGM6-4r-based ELISA, which was applied for surra against cattle and water buffalo and dourine against horse, revealed that the overall sero-prevalence of the diseases in Mongolia was 4.8%. Among them, high sero-prevalences were observed in the central provinces (5.2–11.0%, p < 0.05) of the country. The sero-prevalence was significantly higher in females than in males (6.0% and 4.0%, p < 0.05, respectively) and in non-castrated males (8.4%, p < 0.01) compared with castrated males (3.0%). These results suggested that currently, horse trypanosomoses are a nationwide endemic problem in Mongolia. Knowledge of the nationwide endemic status of non-tsetse-transmitted horse trypanosomoses in Mongolia will be useful to prevent these diseases.

Understanding the role of English language acquisition in developing the socioeconomic status of Vietnam forms the backdrop for this study, which seeks to shed light on the potential benefits of technology-assisted vocabulary learning. Based on this context, this study employed a mixed method of quantitative and qualitative design attempting to investigate the vocabulary development of college students (n=23) who were treated by the use of Memrise (Experimental group) in combination with a traditional teaching method compared to their counterparts (n=24) who were taught by a traditional teaching method (TTM) (Control group) without the use of this application. Students' thoughts about the benefits and challenges of Memrise for their vocabulary learning after the treatment were also investigated. The study yielded noteworthy results on multiple fronts. Quantitatively, data collected from the vocabulary post-tests showed that the experimental group significantly outperformed the control group in their vocabulary scores. Qualitatively, interview data indicated that Memrise positively impacted students' motivation and interest for vocabulary learning, suggesting its effectiveness in addressing the particular problems of engagement and retention in vocabulary acquisition. However, there were logistical challenges, particularly in rural settings. Factors like unstable internet connections and inadequate mobile devices adversely affected the quality of learning experiences with Memrise. By focusing on these elements, this study highlights key pedagogical implications and underscores the value of technological interventions in language learning, offering insights into addressing infrastructure barriers and tailoring e-learning methods to diverse learning environments and needs.

The research aims to investigate the support suppliers’ decision making for Textile and Garment Vietnamese Enterprises based on using association rules mining. In order to increase the competitive advantage over other enterprises, Vietnamese enterprises are gradually shifting their production methods from CMT (Cut-Make-Trim) to FOB (Free On Board), ODM (Original Design Manufacturing), and OBM (Original Brand Manufacturing). One of the important steps for this transformation is to be self-sufficient in sourcing raw materials from selected suppliers with sufficient capabilities, meeting the requirements of price, and CSR (corporate social responsibility) to enable Vietnamese enterprises to enjoy tax incentives. The Vietnamese Enterprises participate in the textile and garment production process through importing raw materials from other countries such as China, Korea, etc. Therefore, the selection supplier issue is very important to them to enhance their business products. This study researches on finding positive ARs and negative ARs in Textile and Garment data domain via three scenarios to define the relationship of alternative criteria for selection suppliers. By using alternative method of ARM (Association Rule Mining), the experimental results show that Textile and Garment enterprises in Vietnam appreciate the CSR factor highly. This is completely in line with the previous survey research on the new factor in the supplier selection criteria of Textile and Garment enterprises in Vietnam. In another word, in this field, the CSR plays as an important role to reflect company’s image and it is the first of decision choice. The other criteria are considered after including Cost, Quality, and Delivery respectively. The remain factors of Service, Capability, Relationship, and Sourcing Country are not highly evaluated via the experiments. With different scenarios along with the association rules mined, they reflect different sets of criteria that textile and garment enterprises use to select their suppliers. Furthermore, the rare rules show situations that are very unlikely but have high reliability in reality. Based on the findings, the factors will show the essentials for suppliers to improve themselves to meet the requirements.

Purpose – By adopting the theory of planned behaviour, this study aims to provide insights into how entrepreneurial education (EE) contributes to the entrepreneurial process, especially entrepreneurial behaviour (EB). Design/methodology/approach – The research used a sample of 2,566 students from 16 universities in Vietnam. The conceptual framework’s coefficient paths underwent testing using structural equation modeling, and the mediation effects were determined using the PROCESS bootstrapping method. Findings – The study revealed that EE directly influences EB in Vietnam, despite having no direct effect on entrepreneurial intention (EI). In addition, EE indirectly influences EI and EB through enhancing attitude towards entrepreneurship and perceived behavioural control. Practical implications – These findings suggest that policymakers should consider the value of investing in EE programmes as a means of promoting EB. Offering students opportunities to engage in real-life entrepreneurial activities, such as business plan competitions, internships with local start-ups and access to mentorship from established entrepreneurs, can play a pivotal role in their translating knowledge into action. This practical approach can significantly contribute to the development of an entrepreneurial ecosystem in Vietnam. Originality/value – While many studies have investigated the influence of EE on EIs, there is a lack of research on the behavioural outcomes of such education. Furthermore, the findings regarding the EE–EI relationship are inconsistent. Therefore, this study provides valuable insights into how EE can foster the intention and behaviour to engage in entrepreneurship.

Sexual stage induction under in vitro conditions is useful for biological and molecular studies of Babesia parasites. Therefore, in the present study, we induced B. ovata tick stages using the chemical inducers: xanthurenic acid (XA), dithiothreitol (DTT) and tris (2-carboxyethyl) phosphine (TCEP) at 27 °C or 37 °C conditions. Cultures at low temperature (27 °C) or treated with XA/TCEP induced a large number of extra-erythrocytic merozoites, which transformed into round shape cells at 12−24 h post-induction (pi). However, typical forms of tick stages (aggregation forms and the spiky forms/ray bodies) were only observed in the cultures treated with 40 mM or 60 mM of DTT during 3−6 h pi. The induced cells were recognized by anti-CCp2 rabbit antisera. DNA content of the cell population treated with 40 mM of DTT was analyzed by imaging flow cytometry at 0, 12 and 48 h pi. The results indicated that the parasite population with diploid-like double DNA content increased at 48 h pi. Our observations on morphological and changes in the DNA content provide useful information for understanding the life cycle of B. ovata under in vitro conditions, which will facilitate further studies on basic biology and the development of transmission blocking vaccines against bovine babesiosis.

This study evaluates the potential for agritourism development in Binh Dinh Province, Central Vietnam. The analytical hierarchical process is used to determine the weights of four factors with 25 measurement criteria to assess agritourism potential. The research results show that Binh Dinh Province has great agritourism potential based on four evaluation factors, including (1) local agricultural tourism resources, (2) the trend for agritourism development and the agritourism market, (3) the local community’s willingness, and (4) the supporting activities of the local government. In particular, the local agricultural tourism resources factor received the highest score, followed by the local community’s willingness and the supporting activities of the local government. The trend for agritourism development and the agritourism market was rated the lowest. Therefore, Binh Dinh has viable opportunities to encourage the growth of agritourism to improve farmers’ livelihoods