Glycosylphosphatidylinositol-anchored micronemal antigen (GAMA) interacts with the band 3 receptor to promote erythrocyte invasion by malaria parasites.

Glycosylphosphatidylinositol-anchored micronemal antigen (GAMA) is an erythrocyte binding protein known to be involved in malarial parasite invasion. Although anti-GAMA antibodies have been shown to block GAMA attachment to the erythrocyte surface and subsequently inhibit parasite invasion, little is known about the molecular mechanisms by which GAMA promotes the invasion process. In this study, LC-MS analysis was performed on the erythrocyte membrane to identify the specific receptor that interacts with GAMA. We found that ankyrin 1 and the band 3 membrane protein showed affinity for GAMA, and characterization of their binding specificity indicated that both Plasmodium falciparum and Plasmodium vivax GAMA bound to the same extracellular loop of band 3 (loop 5). In addition, we show the interaction between GAMA and band 3 was sensitive to chymotrypsin. Furthermore, antibodies against band 3 loop 5 were able to reduce the binding activity of GAMA to erythrocytes and inhibit the invasion of P. falciparum merozoites into human erythrocytes, whereas antibodies against P. falciparum GAMA (PfGAMA)-Tr3 only slightly reduced P. falciparum invasion. The identification and characterization of the erythrocyte GAMA receptor is a novel finding that identifies an essential mechanism of parasite invasion of host erythrocytes.

Long-term immune responses in patients with confirmed novel coronavirus disease-2019: a 9-month prospective cohort study in Shanghai, China.

BACKGROUND: The duration of antibodies against SARS-CoV-2 in Covid-19 patients remains uncertain. Longitudinal serological studies are needed to prevent disease and transmission of the virus. METHODS: In 2020, 414 blood samples were tested, obtained from 157 confirmed Covid-19 patients, in a prospective cohort study in Shanghai. RESULTS: The seropositive rate of IgM peaked at 40.5% (17/42) within 1 month after illness onset and then declined. The seropositive rate of IgG was 90.6% (58/64) after 2 months, remained above 85% from 2 to 9 months and was 90.9% (40/44) after 9 months. Generalized estimating equations models suggested that IgM (P < 0.001) but not IgG significantly decreased over time. Age ≥ 40 years (adjusted odds ratio [aOR] 4.531; 95% confidence interval [CI] 1.879-10.932), and cigarette smoking (aOR 0.344; 95% CI 0.124-0.951) were associated with IgG, and age ≥ 40 years (aOR 2.820; 95% CI 1.579-5.036) was associated with IgM. After seroconversion, over 90% and 75.1% of subjects were estimated to remain IgG-positive 220 and 254 days, respectively. Of 1420 self-reported symptoms questionnaires, only 5% reported symptoms 9 months after onset. CONCLUSIONS: In patients with a history of natural infection, anti-SARS-CoV-2 IgG is long-lived, being present for at least 9 months after illness onset. The long duration of natural immunity can mitigate and eliminate Covid-19 and the ongoing pandemic.

A Chimeric Plasmodium vivax Merozoite Surface Protein Antibody Recognizes and Blocks Erythrocytic P. cynomolgi Berok Merozoites In Vitro.

Research on erythrocytic Plasmodium vivax merozoite antigens is critical for identifying potential vaccine candidates in reducing P. vivax disease. However, many P. vivax studies are constrained by its inability to undergo long-term culture in vitro Conserved across all Plasmodium spp., merozoite surface proteins are essential for invasion into erythrocytes and highly expressed on erythrocytic merozoites, thus making it an ideal vaccine candidate. In clinical trials, the P. vivax merozoite surface protein 1 (PvMSP1-19) vaccine candidate alone has shown to have limited immunogenicity in patients; hence, we incorporate the highly conserved and immunogenic C terminus of both P. vivax merozoite surface protein 8 (PvMSP8) and PvMSP1-19 to develop a multicomponent chimeric protein rPvMSP8+1 for immunization of mice. The resulted chimeric rPvMSP8+1 antibody was shown to recognize native protein MSP8 and MSP1-19 of mature P. vivax schizonts. In the immunized mice, an elevated antibody response was observed in the rPvMSP8+1-immunized group compared to that immunized with single-antigen components. In addition, we examined the growth inhibition of these antibodies against Plasmodium cynomolgi (Berok strain) parasites, which is phylogenetically close to P. vivax and sustains long-term culture in vitro Similarly, the chimeric anti-rPvMSP8+1 antibodies recognize P. cynomolgi MSP8 and MSP1-19 on mature schizonts and showed strong inhibition in vitro via growth inhibition assay. This study provides support for a new multiantigen-based paradigm rPvMSP8+1 to explore potential chimeric vaccine candidates against P. vivax malaria using sister species P. cynomolgi.

Immunogenicity analysis of conserved fragments in Plasmodium ovale species merozoite surface protein 4.

BACKGROUND: There is an urgent need for an effective vaccine to control and eradicate malaria, one of the most serious global infectious diseases. Plasmodium merozoite surface protein 4 (MSP4) has been listed as a blood-stage subunit vaccine candidate for malaria. Infection with Plasmodium ovale species including P. ovale wallikeri and P. ovale curtisi, is also a source of malaria burden in tropical regions where it is sometimes mixed with other Plasmodium species. However, little is known about P. ovale MSP4. METHODS: The msp4 gene was amplified through polymerase chain reaction using genomic DNA extracted from blood samples of 46 patients infected with P. ovale spp. and amplified products were sequenced. Open reading frames predicted as immunogenic peptides consisting of 119 and 97 amino acids of P. ovale curtisi MSP4 (PocMSP4) and P. ovale wallikeri MSP4 (PowMSP4), respectively, were selected for protein expression. Recombinant proteins (rPoMSP4) were expressed in Escherichia coli, purified, analysed, and immunized in BALB/c mice. The specificity of anti-MSP4-immunoglobulin (Ig) G antibodies was evaluated by Western blot and enzyme-linked immunosorbent assays, and cellular immune responses were analysed via lymphocyte proliferation assays. RESULTS: Full peptide sequences of PocMSP4 and PowMSP4 were completely conserved in all clinical isolates, except in the epidermal growth factor-like domain at the carboxyl terminus where only one mutation was observed in one P. o. wallikeri isolate. Further, truncated PoMSP4 segments were successfully expressed and purified as ~ 32 kDa proteins. Importantly, high antibody responses with end-point titres ranging from 1:10,000 to 1:2,560,000 in all immunized mouse groups were observed, with high IgG avidity to PocMSP4 (80.5%) and PowMSP4 (92.3%). Furthermore, rPocMSP4 and rPowMSP4 cross-reacted with anti-PowMSP4-specific or anti-PocMSP4-specific antibodies. Additionally, anti-PoMSP4 IgG antibodies showed broad immuno-specificity in reacting against rPoMSP1 and rPoAMA1. Lastly, PocMSP4- and PowMSP4-immunized mice induced cellular immune responses with PocMSP4 (36%) and PowMSP4 cells (15.8%) during splenocyte proliferation assays. CONCLUSION: Findings from this study suggest conservation in PoMSP4 protein sequences and high immunogenicity was observed in rPoMSP4. Furthermore, induction of immune responses in PocMSP4- and PowMSP4-immunized mice informed that both humoral and cellular immune responses play crucial roles for PoMSP4 in protection.

[Effect of prenatal BDE-99 exposure on the maternal thyroid hormones levels and development of rat offspring].

OBJECTIVE: To investigate the effects of prenatal exposure to pentabromodiphenyl ether(BDE-99) on maternal serum thyroid hormone levels, as well as birth weight and anal-genital development in rat offspring. METHODS: Pregnant SD rats were randomly treated with BDE-99(0. 2, 2 and 20 mg/kg) or corn oil on gestational days 1-19. Maternal serum were collected from tail vein on the gestational day 11 and day 19, serum levels of TSH, TT4, FT4, TT3 and FT3 were measured. The weight of offspring was measured at postnatal day 3, 9, 15, 21 and 27, anorectal-genital spacing was measured at postnatal day 21. RESULTS: The levels of TT3 and FT3 in maternal serum of 2 mg/kg and 20 mg/kg groups were lower than those of control group at gestational day 11. At gestational day 19, TT4 levels in maternal serum were significantly lower than those in control group, and TSH levels of 20 mg/kg group were lower than those in control group. The body weight of female offspring in all dose groups was lower than that of control group on the postnatal day 27, and the anal-genital distance of male offspring in the 20 mg/kg dose group was significantly lower than that of the control group on the postnatal day 21. CONCLUSION: Prenatal exposure to BDE-99 may disrupt the maternal thyroid hormone levels, and cause the offspring's weight loss and shortened anal-genital spacing.

Anisotropic Weyl fermions from the quasiparticle excitation spectrum of a 3D Fulde-Ferrell superfluid.

Weyl fermions, first proposed for describing massless chiral Dirac fermions in particle physics, have not been observed yet in experiments. Recently, much effort has been devoted to explore Weyl fermions around band touching points of single-particle energy dispersions in certain solid state materials (named Weyl semimetals), similar as graphene for Dirac fermions. Here we show that such Weyl semimetals also exist in the quasiparticle excitation spectrum of a three-dimensional spin-orbit-coupled Fulde-Ferrell superfluid. By varying Zeeman fields, the properties of Weyl fermions, such as their creation and annihilation, number and position, as well as anisotropic linear dispersions around band touching points, can be tuned. We study the manifestation of anisotropic Weyl fermions in sound speeds of Fulde-Ferrell fermionic superfluids, which are detectable in experiments.

The assessment of transmission efficiency and latent infection period in asymptomatic carriers of SARS-CoV-2 infection.

Few studies have focused on the transmission efficiency of asymptomatic carriers of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Our follow-up study was performed on 147 asymptomatic carriers in Anhui Province. Of these, 50.0% were male, 50.3% were older than 40 years, 43.8% were farmers, and 68.7% were from the north of Anhui Province. 16 of the 147 asymptomatic carriers developed symptoms in the following 14 days of isolated observation, and were subsequently diagnosed as confirmed cases. The possible latent infection period was found to range from 1-5 days before onset, with a median time of 2 days. The second attack rate for the 16 confirmed cases who had transferred from being asymptomatic carriers was 9.7% (23/236 close contacts), while for the 131 asymptomatic carriers the rate was 2.6% (24/914 close contacts), showing a significant difference in second attack rate between the two groups (p＜0.001). Our study indicated that COVID-19 cases are contagious during the incubation period, and that close contact screening should be extended to include the incubation period. Our results also showed that the transmission efficiency for asymptomatic carriers was lower than that for confirmed case.

Low genetic diversity and strong immunogenicity within the apical membrane antigen-1 of plasmodium ovale spp. imported from africa to china.

Malaria is still an important challenge for global public health because of its extensive mortality and morbidity. Plasmodium ovale is mainly distributed in tropical regions of Africa and Asia. it includes two distinct ovale malaria species, which are P. ovale curtisi and P. ovale wallikeri. Apical membrane antigen-1 (AMA-1) is an asexual blood-stage protein which is essential for Plasmodium. Thus far, no study on gene polymorphism and immunogenicity of P. ovale AMA-1 (PoAMA-1) has been conducted. Amplified poama1 gene products from 14 P ovale curtisi samples and 12 P ovale wallikeri samples imported from Africa to Jiangsu Province, China were sequenced and their polymorphisms were analyzed. We expressed recombinant PoAMA-1 (rPoAMA-1, 53 kDa) proteins in an E. coli expression system and evaluated immune responses against the rPoAMA-1 in BALB/c mice. We identified a synonymous mutation in nucleotide position 333 of the pocama-1 gene and powama-1 did not reveal any variation. The humoral and cellular immune responses to rPoAMA-1 were evaluated using enzyme-linked immunosorbent assay (ELISA) and flow cytometry. rPoAMA-1-immunized mice produced specific antibodies as verified by immunoblotting. The rPoAMA-1 induced high antibody titers (1: 640,000), and had high avidity indexes (an average of 78.63% and 83.40%). The antibodies also recognized the native proteins, namely, crude antigen from blood stages. Cross-reactivity between rPocAMA-1 and rPowAMA-1 was observed. Moreover, rPoAMA-1 s induced interferon (IFN)-gamma-secreting cells in mice and increased lymphocyte proliferation response. Low genetic diversity was observed in poama-1 from the Jiangsu Province imported malaria cases, and further studies conclusively showed its strong immunogenicity. Significant cross-reactivity was found between rPocAMA-1 and rPowAMA-1, suggesting that a single PoAMA-1 antigen could be used to diagnose P. ovale curtisi or P. ovale wallikeri patient simultaneously. However, further evaluation needs to be carried out to validate the potential and limitations of PoAMA-1 as a candidate vaccine.

Immunogenicity analysis of genetically conserved segments in Plasmodium ovale merozoite surface protein-8.

BACKGROUND: Plasmodium ovale is widely distributed across tropical regions and has two closely related but distinct species, namely P. ovale curtisi and P. ovale wallikeri. Combining genetic characterization with the immunogenicity of merozoite surface protein-8 (MSP-8) supports considering MSP-8 as a candidate target for blood-stage vaccines against malaria. However, no previous studies have focused on characterizing the genetic diversity and immunogenicity of PoMSP-8. METHODS: Blood samples were collected from 42 patients infected with P. ovale. The patients were migrant workers returning to the Jiangsu Province from Africa; genomic DNA was extracted from their blood samples for sequencing and protein expression. The recombinant PoMSP-8 (rPoMSP-8) proteins were expressed and purified to assess their immune responses in BALB/c mice. RESULTS: The sequences of the P. ovale curtisi and P. ovale wallikeri msp8 genes were completely conserved in each isolate. The rPoMSP-8 proteins were successfully expressed and purified as ~70 kDa proteins. Antibodies raised against rPoMSP-8 in mice showed appropriate immunoreactivity, as evidenced by immunoblotting. These specific antibodies were detected at day 7 post-immunization, and their levels increased throughout the whole immunization period. rPoMSP-8-raised antibodies had high endpoint titers (1:5,120,000) and high avidity (PocMSP-8: 94.84%, PowMSP-8: 92.69%). Cross-reactivity between rPocMSP-8 and rPowMSP-8 was observed with each anti-PoMSP8-specific antibody. CONCLUSIONS: Remarkable conservation and high immunogenicity was observed in both rPoMSP-8s. Intriguingly, cross-reaction between rPocMSP-8 and rPowMSP-8 was detected, suggesting that a single PoMSP8-based construction might be applicable for both species.

Limited genetic diversity of N-terminal of merozoite surface protein-1 (MSP-1) in Plasmodium ovale curtisi and P. ovale wallikeri imported from Africa to China.

BACKGROUND: Plasmodium merozoite surface protein-1 (MSP-1) is released into the bloodstream during merozoite invasion, and thus represents a crucial malarial vaccine target. Although substantial research effort has been devoted to uncovering the genetic diversity of MSP-1 for P. falciparum and P. vivax, there is minimal information available regarding the genetic profiles and structure of P. ovale. Therefore, the aim of the present study was to determine the extent of genetic variation among two subspecies of P. ovale by characterizing the MSP-1 N-terminal sequence at the nucleotide and protein levels. METHODS: N-terminal of MSP-1 gene were amplified from 126 clinical samples collected from imported cases of malaria in migrant workers returning to Jiangsu Province from Africa using a conventional polymerase chain reaction (PCR) assay. The PCR products were then sequenced and analyzed using the GeneDoc, MegAlign, MEGA7 and DnaSP v.6 programs. RESULTS: The average pairwise nucleotide diversities (π) of P. ovale curtisi and P. ovale wallikeri MSP-1 genes (pomsp1) were 0.01043 and 0.01974, respectively, and the haplotype diversity (Hd) were 0.746 and 0.598, respectively. Most of the nucleotide substitutions detected were non-synonymous, indicating that the genetic variations of pomsp1 were maintained by positive diversifying selection, thereby suggesting their role as a potential target of a protective immune response. Amino acid substitutions of P. ovale curtisi and P. ovale wallikeri MSP-1 could be categorized into five and three unique amino acid variants, respectively. CONCLUSIONS: Low mutational diversity was observed in pomsp1 from the Jiangsu Province imported malaria cases; further studies will be developed such as immunogenicity and functional analysis.

Topological Anderson insulator.

Disorder plays an important role in two dimensions, and is responsible for striking phenomena such as metal-insulator transition and the integral and fractional quantum Hall effects. In this Letter, we investigate the role of disorder in the context of the recently discovered topological insulator, which possesses a pair of helical edge states with opposing spins moving in opposite directions and exhibits the phenomenon of quantum spin Hall effect. We predict an unexpected and nontrivial quantum phase termed "topological Anderson insulator," which is obtained by introducing impurities in a two-dimensional metal; here disorder not only causes metal-insulator transition, as anticipated, but is fundamentally responsible for creating extended edge states. We determine the phase diagram of the topological Anderson insulator and outline its experimental consequences.

Finite size effects on helical edge states in a quantum spin-Hall system.

The hallmark of the spin-Hall insulator is the presence of gapless edge states of different spins moving in opposite directions. Through analytical solutions in a model calculation for a strip of finite width, we find that edge states on the two sides can couple together to produce a gap in the spectrum, destroying the quantum spin-Hall effect. The application of a magnetic field can however modify and even remove the gap by shifting the momenta of the edge states relative to each other.