Investigation of RBM10 mutation and its associations with clinical and molecular characteristics in EGFR-mutant and EGFR-wildtype lung adenocarcinoma.

Background: RBM10 is commonly mutated in lung adenocarcinoma (LUAD). However, its role in the pathogenesis of LUAD remains undefined. EGFR-mutant LUAD represents a distinct subset of non-small cell lung cancer (NSCLC). The function of RBM10 in tumor pathogenesis is supposed to differ between EGFR-mutant and EGFR-wt LUAD. This study aimed to interrogate the prevalence of RBM10 mutation in a large cohort of Chinese patients with LUAD and investigate the association of RBM10 mutation with clinical and molecular characteristics of EGFR-mutant and EGFR-wt LUAD. Methods: Tumor sequencing data from 2848 Chinese patients with LUAD were retrospectively reviewed and analyzed. The prevalence of RBM10 was also compared with other three cohorts: OrigMed (n = 1222), MSKCC (n = 1267), and TCGA (n = 566). The associations of RBM10 mutation with clinical and molecular characteristics were assessed. An external cohort of 182 patients with LUAD who received PD-1 inhibitor were used to investigate the association of RBM10 mutation with clinical outcomes upon immunotherapy. Results: Our cohort showed a higher prevalence of RBM10 in EGFR-mutant LUAD than in EGFR-wt LUAD (14.8 % vs. 6.5 %, p < 0.001). The enrichment of RBM10 mutations in EGFR-mutant LUAD was also seen in another Chinese cohort (OrigMed: 14.9 % vs. 7.8 %, p < 0.001), but not in the two western cohorts (MSKCC: 7.4 % vs. 9.5 %, p = 0.272; TCGA: 8.1 % vs. 6.7 %, p = 0.624). RBM10 mutations co-occurred more frequently with EGFR L858R mutations (23.7 %) than with other types of EGFR mutations (19 del: 7.7 %; other: 7.1 % in others, p < 0.001). In EGFR-mutant LUAD, RBM10 mutations were more commonly found in stage I (18.2 %) and II (21.8 %) vs. stage III (9.4 %) and IV (11.3 %) tumors (p < 0.001). The proportion of PD-L1 positive expression in EGFR-mutant LUAD with concomitant RBM10 mutation was not different from that those without RBM10 mutations (41.8 % vs. 47.9 %, p = 0.566). In contrast, RBM10 mutation occurred more frequently in EGFR-wt LUAD at stage II-IV (stage II: 12.0 %, stage III: 8.7 %, stage IV: 6.6 %) than at stage I (2.8 %). EGFR-wt LUAD with concomitant RBM10 mutations had higher proportions of PD-L1 expression positivity (78.9 % vs. 61.9 %, p = 0.014) and higher tumor mutational load (8.97 vs. 2.99 muts/Mb, p < 0.001) than those without. Patients with EGFR-wt LUAD who also harbored RBM10 loss of function (LOF) mutations had a longer median PFS upon immunotherapy than those with RBM10 non-LOF mutations (7.15 m vs. 2.60 m, HR = 4.83 [1.30-17.94], p = 0.010). Conclusion: We comprehensively investigated RBM10 mutations in a Chinese cohort with LUAD. Compared to western cohorts, a significant enrichment of RBM10 mutations in EGFR-mutant LUAD compared to EGFR-wildtype LUAD in the Chinese population. RBM10 mutation shows different associations with clinical and molecular characteristics between EGFR-mutant and EGFR-wt LUAD, suggesting a divergent mechanism between these two subsets via which RBM10 deficiency contribute to tumor pathogenesis. The findings contribute to our understanding of the molecular landscape of LUAD and highlight the importance of considering population-specific factors in cancer genomics research.

Does sadness bring myopia: an intertemporal choice experiment with college students.

Introduction: While economics often interprets individual intertemporal choice preferences through the rationality assumption of utility maximization, the reality is that as emotional beings, individuals' preferences for intertemporal behavior are much more diverse and inconsistent. Prior research has predominantly focused on positive or negative emotions based on prospect theory, such as anxiety, anger, disgust, and depression. However, there has been relatively little research on how sadness affects individuals' preferences for immediate and future rewards. Methods: In this study, 170 college students are recruited as participants, and their emotions are primed with a video before engaging in an intertemporal task. Covariance analysis and logit regression model are established to examine the main and interactive effects of sadness on individuals' immediate reward preferences. Results: The findings reveal that sadness led individuals to prefer smaller immediate rewards, demonstrating a more myopic behavioral pattern, but didn't affect time discount rate. As the reward baseline increases, sadness's impact on immediate reward preferences is more pronounced, exacerbating individuals' myopic behavior. Discussion: In conclusion, these findings underscore the importance of considering emotional states in economic decision-making models and suggest avenues for future research to explore the complex dynamics of emotions and intertemporal choices.

The Role of Gluten in Food Products and Dietary Restriction: Exploring the Potential for Restoring Immune Tolerance.

Wheat is extensively utilized in various processed foods due to unique proteins forming from the gluten network. The gluten network in food undergoes morphological and molecular structural changes during food processing, affecting the final quality and digestibility of the food. The present review introduces the formation of the gluten network and the role of gluten in the key steps of the production of several typical food products such as bread, pasta, and beer. Also, it summarizes the factors that affect the digestibility of gluten, considering that different processing conditions probably affect its structure and properties, contributing to an in-depth understanding of the digestion of gluten by the human body under various circumstances. Nevertheless, consumption of gluten protein may lead to the development of celiac disease (CD). The best way is theoretically proposed to prevent and treat CD by the inducement of oral tolerance, an immune non-response system formed by the interaction of oral food antigens with the intestinal immune system. This review proposes the restoration of oral tolerance in CD patients through adjunctive dietary therapy via gluten-encapsulated/modified dietary polyphenols. It will reduce the dietary restriction of gluten and help patients achieve a comprehensive dietary intake by better understanding the interactions between gluten and food-derived active products like polyphenols.

UV-B irradiation-activated E3 ligase GmILPA1 modulates gibberellin catabolism to increase plant height in soybean.

Plant height is a key agronomic trait that affects yield and is controlled by both phytohormone gibberellin (GA) and ultraviolet-B (UV-B) irradiation. However, whether and how plant height is modulated by UV-B-mediated changes in GA metabolism are not well understood. It has not been reported that the E3 ubiquitin ligase Anaphase Promoting Complex/Cyclosome (APC/C) is involved in the regulation of plant growth in response to environmental factors. We perform a forward genetic screen in soybean and find that a mutation in Glycine max Increased Leaf Petiole Angle1 (GmILPA1), encoding a subunit of the APC/C, lead to dwarfism under UV-B irradiation. UV-B promotes the accumulation of GmILPA1, which ubiquitinate the GA catabolic enzyme GA2 OXIDASE-like (GmGA2ox-like), resulting in its degradation in a UV-B-dependent manner. Another E3 ligase, GmUBL1, also ubiquitinate GmGA2ox-like and enhance the GmILPA1-mediated degradation of GmGA2ox-like, which suggest that GmILPA1-GmGA2ox-like module counteract the UV-B-mediated reduction of bioactive GAs. We also determine that GmILPA1 is a target of selection during soybean domestication and breeding. The deletion (Indel-665) in the promoter might facilitate the adaptation of soybean to high UV-B irradiation. This study indicates that an evolutionary GmILPA1 variant has the capability to develop ideal plant architecture with soybean cultivars.

Electron Transport in Organic Photovoltaic Acceptor Materials: Improving the Carrier Mobilities by Intramolecular and Intermolecular Modulations.

High carrier mobility is beneficial to increase the active-layer thickness while maintaining a high fill factor, which is crucial to further improve the light harvesting and organic photovoltaic efficiency. The aim of this Perspective is to elucidate the electron transport mechanisms in prototypical non-fullerene (NF) acceptors through our recent theoretical studies. The electron transport in A-D-A small-molecule acceptors (SMAs), e.g., ITIC and Y6, is mainly determined by end-group π-π stacking. Relative to ITIC, the angular backbone along with more flexible side chains leads to Y6 having a closer stacking and enhanced intermolecular electronic connectivity. For polymerized rylene diimide acceptors, to achieve high electron mobilities, they need to simultaneously increase intramolecular and intermolecular connectivity. Finally, finely tuning the π-bridge modes to enhance intramolecular superexchange coupling is essential to develop novel polymerized A-D-A SMAs.

Kinetics of thermal degradation of raw lacquer enhanced by formaldehyde urea prepolymer.

In this study, formaldehyde-urea prepolymer (FUP) were synthesized, which were used to modify the raw lacquer (RL) and this composition named LF, while the basic properties of the RL were tested. Thermal gravimetric (TG) analysis and scanning electron microscopy (SEM) were used to analyze the degradative characteristics and the surface morphology of RL before and after modification. The result indicated that FUP can significantly improve the performance of RL. The drying time of the LF is significantly shortened, the gloss, the pencil hardness, and the impact performance are significantly enhanced at the same time. TG analysis and thermal decomposition kinetics analysis illustrated that the thermal stability and the activation energy of LF2 were stronger than that of RL. In addition, SEM analysis illustrated that the surface smoothness of RL were also improved.

Effects of childhood trauma on aggressive behaviors and hippocampal function: the modulation of COMT haplotypes.

Background: Aggression is a commonly hostile behavior linked to the hippocampal activity. Childhood trauma (CT) exposure has been associated with altered sensitization of the hypothalamic-pituitary-adrenal (HPA) axis and hippocampal volume，which could increase violent aggressive behaviors. Additionally, Catechol-O-methyltransferase (COMT), the major dopamine metabolism enzyme, is implicated in stress responsivity, including aggression. Hence, CT exposure may affect aggression through the effect on the hippocampal function, which might also be modulated by the COMT variations. Objectives: This study examined whether both CT and haplotypes of COMT moderate hippocampal function and thus affect human aggressive behavior. Methods: We obtained bilateral hippocampal functional connectivity maps using resting state functional magnetic resonance imaging (MRI) data. COMT haplotype estimation was performed using Haploview 4.2 and PHASE 2.1. Then we constructed a moderated mediation model to study the effect of the CTQ × COMT on aggressive behavior. Results: Three major haplotypes were generated from thirteen single nucleotide polymorphisms (SNPs) within the COMT gene and formed three haplotypes corresponding to high, medium, and low enzymatic activity of COMT. The results showed interactive relationships between the Childhood Trauma Questionnaire (CTQ) and COMT with respect to the functional connectivity (FC) of the bilateral hippocampus (HIP)-orbital frontal cortex (OFC). Specifically, CT experience predicted lower negative HIP-OFC coupling in the APS and HPS haplotypes corresponding to the medium and high enzymatic activity of COMT, but greater FC in the LPS haplotypes corresponding to the low enzymatic activity. We also observed a conditional mediation effect of the right HIP-OFC coupling in the link between COMT and aggressive behavior that was moderated by CT experience. Conclusions: These results suggest that CT and COMT have a combined effect on aggressive behavior through hippocampal function. This mediation analysis sheds light on the influence of childhood experience on aggressive behavior in different genetic backgrounds.

Efficacy of osimertinib in a metastatic lung adenocarcinoma patient harboring somatic EGFR delL747_S752 and germline BIM deletion polymorphism: a case report and literature review.

Background: Epidermal growth factor receptor (EGFR) exon 19 deletion (19del) and the exon 21 L858R point mutation are the most established predictive factors for the efficacy of EGFR-tyrosine kinase inhibitor (TKI) in patients with non-small cell lung cancer (NSCLC). To date, more than 50 subtypes of EGFR 19dels have been documented in NSCLC. Evidence suggests that different subtypes of 19dels exhibit different survival outcomes to EGFR-TKI treatment. Whether patients harboring EGFR Leu747_Ser752 deletion (delL747_S752) as an uncommon subtype of 19dels benefit from EGFR-TKIs has not been investigated. BIM (B-cell chronic lymphocytic leukemia/lymphoma-like 1) deletion polymorphism is common in East Asian with EGFR-mutant NSCLC. Currently, the predictive role of BIM deletion polymorphism in patients with EGFR-mutant NSCLC treated with osimertinib remains debatable. Case Description: A 34-year-old female patient was diagnosed with stage IV lung adenocarcinoma (LUAD) harboring somatic EGFR del L747_S752 and germline BIM deletion polymorphism in August 2018. She obtained benefit from the front-line treatment of osimertinib lasting for 8 months. After progression from osimertinib, the patient received bevacizumab combined with platinum-doublet chemotherapy, stereotactic radiosurgery plus osimertinib and crizotinib, anlotinib, and a programmed cell death-1 inhibitor sintilimab plus bevacizumab and docetaxel. She succumbed to her disease in June 2020 with an overall survival of 23 months. Conclusions: Our work suggests that osimertinib might be a compromised treatment option for NSCLC patients with somatic EGFR delL747_S752 and germline BIM deletion polymorphism. Development of more effective regimens are needed for this small subset of NSCLCs.

Lung microbiome in children with hematological malignancies and lower respiratory tract infections.

Background: Respiratory infectious complications remain a major cause of morbidity and mortality in children with hematological malignancies. Knowledge regarding the lung microbiome in aforementioned children is limited. Methods: A prospective cohort was conducted, enrolling 16 children with hematological malignancies complicated with moderate-to-severe lower respiratory tract infections (LRTIs) versus 21 LRTI children with age, gender, weight, and infection severity matched, with no underlying malignancies, to evaluate the lung microbiome from bronchoalveolar lavage fluid samples in different groups. Results: The lung microbiome from children with hematological malignancies and LRTIs showed obviously decreased α and ß diversity; increased microbial function in infectious disease:bacteria/parasite; drug resistance:antimicrobial and human pathogenesis than the control group; a significantly reduced proportion of Firmicutes, Bacteroidota, Actinobacteriota; increased Proteobacteria at the phylum level; and distinctly elevated Parabacteroides, Klebsiella, Grimontia, Escherichia_Shigella, unclassified_Enterobacteriaceae at the genus level than the control group. Furthermore, it was revealed that α diversity (Shannon), ß diversity (Bray-Curtis dissimilarity), Proteobacteria at the phylum level, and unclassified_Enterobacteriaceae and Escherichia_Shigella at the genus level were significantly negatively associated with hospitalization course whereas Firmicutes at the phylum level was established positively correlated with the hospitalization course. Conclusions: Children with hematological malignancies and LRTIs showed obviously decreased α and ß diversity, significantly increased function in infectious disease pathogenesis, antimicrobial drug resistance, and unfavorable environment tolerance. Moreover, α diversity (Shannon), ß diversity (Bray-Curtis dissimilarity), and Proteobacteria may be used as negative correlated predictors for hospitalization course in these children whereas Firmicutes may be utilized as a positive correlated predictor.

Interleukin-6 in blood and bronchoalveolar lavage fluid of hospitalized children with community-acquired pneumonia.

Background: Host biomarkers and cytokines help in the prediction of disease severity in adults with community-acquired pneumonia (CAP). Accurate assessment of pathogens and disease severity is essential to clinical decision-making. There are few validated prognostic tools in blood and bronchoalveolar lavage for children with CAP to assist with proper decision and management. Methods: We performed a retrospective study of 118 children under 18 years of age, hospitalized for CAP with bronchoalveolar lavage management within the first 2 days. The primary outcome was disease severity: mild (with no complications), moderate (with mild to moderate complications), and severe (with severe complications). Comparison and performance analysis of biomarkers and cytokines in the blood or bronchoalveolar lavage fluid (BALF) across different severity categories/different pathogens were performed. Results: Analysis of 118 CAP cases revealed significant differences in the BALF levels of IL-6 (p = 0.000), CRP (p = 0.001), and ESR (p = 0.004) across different severity categories, while BALF IL-6 level was indicated as the best indicator to discriminate mild from moderate-to-severe cases with highest AUC (0.847, 95% CI: 0.748-0.946), fair sensitivity (0.839), and specificity (0.450), and severe from non-severe cases with highest AUC (0.847), sensitivity (0.917), and specificity (0.725). ALL biomarkers and cytokines exhibited no significant differences across different pathogen categories (p > 0.05), while BALF IL-6 (p = 0.000), blood ANC (p = 0.028), and ESR (p = 0.024) levels were obviously different in comparison to single Mycoplasma pneumoniae (MP)-, bacteria-, or virus-positive group vs. non-group. Blood CRP (r = 0.683, p = 0.000) and ESR (r = 0.512, p = 0.000) levels revealed significant correlation with the hospitalization course (HC). Among all the BALF cytokines, only BALF IL-6 showed a significant difference (p = 0.004, p < 0.01) across different severity categories, with good performance for predicting CAP severity in hospitalized children (AUC = 0.875, P = 0.004). Blood IL-6 and BALF IL-6 levels showed no significant correlation; in addition, BALF IL6 was better at predicting CAP severity in hospitalized children (AUC = 0.851, p = 0.011, p < 0.05) compared to blood IL-6. Conclusion: BALF IL-6 and blood CRP levels, and ESR may have the ability for discriminating disease severity in hospitalized children with CAP, whereas WBC count and ANC have limited ability. No biomarkers or cytokines seemed to have the ability to predict the pathogen category, while BALF IL-6, blood ANC, and ESR may assist in the diagnosis of single MP, bacteria, and virus infections, respectively.

ATIC facilitates cell growth and migration by upregulating Myc expression in lung adenocarcinoma.

5-Aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (ATIC), a catalysing enzyme in the de novo purine biosynthetic pathway, has been previously reported to be upregulated and to participate in myeloma and hepatocellular carcinoma progression. In the present study, by using bioinformatics technology, a higher ATIC expression was identified in lung adenocarcinoma (LUAD) tissues than in normal tissues, and ATIC expression was found to be positively associated with Myc expression in LUAD tissues. In addition, the role of ATIC in modulating the growth and migration of LUAD cells was explored and the involvement of Myc was revealed. ATIC expression in 56 paired LUAD and tumour adjacent non-cancerous tissues was assessed using reverse transcription-quantitative PCR and western blot analysis. Pearson's correlation analysis was applied to evaluate the correlation between ATIC and Myc expression levels in LUAD tissues. A rescue experiment was performed to explore the role of ATIC/Myc in regulating the growth, migration and invasion of HCC827 and NCI-H1435 cells. It was demonstrated that ATIC was overexpressed in LUAD tissues, particularly in advanced-stage LUAD, and was predicted to be associated with an advanced TNM stage, a higher lymph node metastasis rate, poor tissue differentiation and a lower overall survival rate. ATIC overexpression promoted cell growth, migratory and invasive capacities, whereas this effect was abrogated by Myc knockdown in the HCC827 and NCI-H1435 cells. On the whole, the present study demonstrates that ATIC promotes LUAD cell growth and migration by increasing Myc expression.

Organic Solar Cells with 18% Efficiency Enabled by an Alloy Acceptor: A Two-in-One Strategy.

The trade-off between the open-circuit voltage (Voc ) and short-circuit current density (Jsc ) has become the core of current organic photovoltaic research, and realizing the minimum energy offsets that can guarantee effective charge generation is strongly desired for high-performance systems. Herein, a high-performance ternary solar cell with a power conversion efficiency of over 18% using a large-bandgap polymer donor, PM6, and a small-bandgap alloy acceptor containing two structurally similar nonfullerene acceptors (Y6 and AQx-3) is reported. This system can take full advantage of solar irradiation and forms a favorable morphology. By varying the ratio of the two acceptors, delicate regulation of the energy levels of the alloy acceptor is achieved, thereby affecting the charge dynamics in the devices. The optimal ternary device exhibits more efficient hole transfer and exciton separation than the PM6:AQx-3-based system and reduced energy loss compared with the PM6:Y6-based system, contributing to better performance. Such a "two-in-one" alloy strategy, which synergizes two highly compatible acceptors, provides a promising path for boosting the photovoltaic performance of devices.

Highly Efficient Deamidation of Wheat Gluten by Glucose-Citric Acid-Based Natural Deep Eutectic Solvent: A Potential Effective Reaction Media.

An efficient technique using citric acid and glucose based natural deep eutectic solvent (G-C-NADES) was developed to obtain ultrahigh deamidated wheat gluten (UDWG) (deamidation degree (DD) > 90%). FTIR and 1H NMR indicated intensive hydrogen bonds (HBs) in G-C-NADES supermolecules. Quantum chemical calculations and molecular dynamic simulations demonstrated that 10 wt % diluted G-C-NADES still had a myriad of HBs. Physicochemical results showed UDWG had DD up to 92.45% after G-C-NADES deamidation, that is, 22% higher than citric-acid-DWG with a weak degree of hydrolysis (1.75%). Conformational characterization demonstrated the obvious conversion from α-helix to ß-sheet via FTIR, the least amount of disulfide bonds by Raman spectra, and more exposure of tryptophan residues by fluorescence measurement for UDWG. It is proven that enhanced accessible conformation of WG reached with HBs of G-C-NADESs could contribute to the improvement on nucleophilic attack of deamidation, declaring that G-C-NADES might be a potential solvent for obtaining an ultrahigh deamidation for WG to successfully guarantee the safety of wheat gluten based cereal food regarding to lowering its allergy.

Improved stabilization of coix seed oil in a nanocage-coating framework based on gliadin-carboxymethyl chitosan-Ca2.

Coix seed oil (CSO) is easily suffered functional-loss by oxidation and hydrothermal-treatment. The environmental stable nanocage-coating-CSO particles (OGC-Ca) by the frameworks consist of gliadins, carboxymethyl chitosan (CMCS) and Ca2+ were investigated. Results showed Ca2+ was the key controller for fabricating this nanocage-coating-frameworks, bridging macromolecule-chains with electrostatic interaction and hydrogen bonds, detected by FTIR, CD, DSC and XRD. SEM displayed new-formed velvet-like twigs after cross-linking CMCS to gliadins. Ca2+ assisted the nanocage-coating by significant down-sizing conversion OGC to OGC-Ca with consumption of twigs. OGC-Ca displayed a good stability towards heat (60-80 °C, 0-80 min), pH (3-8), NaCl (0-0.5 mM), storage (4/25 °C, 12 days), and a reduce of the pre-oxidation value of CSO in water and the improved controlled release of CSO in simulated GI tract. It illustrated GC-Ca frameworks would be a suitable delivery carrier for the CSO like pharmaceuticals and nutraceuticals for the food or medical use.

New Synthetic Approaches to N-Aryl and π-Expanded Diketopyrrolopyrroles as New Building Blocks for Organic Optoelectronic Materials.

Diketopyrrolopyrrole (DPP) as a building block has been intensively investigated for organic semiconductors and light emitting materials. The synthesis of N-aryl DPPs remains challenging. Herein, we firstly report a new easily handled synthetic method toward N-aryl DPPs through H-DPP with diaryliodonium salt in the presence of CuI, which shows broad reaction scope. Sixteen N-aryl DPPs, including phenyl, furan and thiophene as flanking aromatic groups, were synthesized with yields up to 78 %. These N-aryl DPPs are fluorescent in both solutions and solid states, with quantum yields up to 96 % and 40 %, respectively. Moreover, we show that the reaction between H-DPP and diaryliodonium salt can lead to π-expanded DPPs by using Pd(OAc)2 as catalyst. Nine π-expanded DPPs were obtained in 27-61 % yields. These π-expanded DPPs exhibit good semiconducting properties with hole mobility of 0.71 cm2 V-1 s-1 , demonstrating that they are useful building blocks for high performance organic semiconductors.

Ac-HSP20 Is Associated With the Infectivity and Encystation of Acanthamoeba castellanii.

Acanthamoeba castellanii is a pathogenic and opportunistic free-living amoeba that causes Acanthamoeba keratitis (AK) and granulomatous amebic encephalitis (GAE) in immunocompromised individuals. The biological and pathogenic characterizations behind this opportunistic protozoan is not fully understood. This study aimed to determine the biological functions of heat shock protein (HSP)-20 of A. castellanii (Ac-HSP20) involved in the maintenance of life cycle and the infectivity of A. castellanii. Immunoscreening A. castellanii cDNA library with A. castellanii infected rabbit sera identified three positive clones, one of them was a putative heat shock protein (Ac-HSP20). The recombinant 23 kDa Ac-HSP20 protein (rAc-HSP20) was successfully expressed in Escherichia coli BL21 (DE3) and purified using metal affinity chromatography. The rabbits immunized with rAc-HSP20 produced high titer antibody (1:25,600). Immunolocalization with the antibody identified the expression of native Ac-HSP20 on the surface of both A. castellanii trophozoites and cysts. Further, Western blot with antibody identified that the expression of native Ac-HSP20 was 7.5 times higher in cysts than in trophozoites. Blocking Ac-HSP20 on the membrane of trophozoites with specific antibody or silencing Ac-hsp20 gene transcription by siRNA inhibited their transformation into cysts at the early stage but returned to normal at the late stage by stimulating the transcription of Ac-hsp20. Incubation of trophozoites with anti-Ac-HSP20 IgG increased macrophage-involved phagocytosis to the protozoa and inhibited trophozoite infectivity on the cornea of rabbits compared with that without antibody. Our study provides that Ac-HSP20 is a surface antigen involved in the encystation and infectivity of A. castellanii and thus an important target for vaccine and drug development.

Vaccination with Bivalent DNA Vaccine of α1-Giardin and CWP2 Delivered by Attenuated Salmonella typhimurium Reduces Trophozoites and Cysts in the Feces of Mice Infected with Giardia lamblia.

BACKGROUND: Giardia lamblia is one of the most common infectious protozoans in human that may cause diarrhea in travelers. Searching for antigens that induced effectively protective immunity has become a key point in the development of vaccine against giardiasis. METHODOLOGY/PRINCIPAL FINDINGS: Mice vaccinated with G. lamblia trophozozite-specific α1-giardin DNA vaccine delivered orally by attenuated Salmonella typhimurium SL7027 elicited 74.2% trophozoite reduction, but only 28% reduction in cyst shedding compared with PBS buffer control. Oral vaccination with Salmonella-delivered cyst-specific CWP2 DNA produced 89% reduction in cysts shedding in feces of vaccinated mice. Significantly, the mice vaccinated with Salmonella-delivered bivalent α1-giardin and CWP2 DNA vaccines produced significant reduction in both trophozoite (79%) and cyst (93%) in feces of vaccinated mice. This parasite reduction is associated with the strong local mucosal IgA secretion and the IgG2a-dominant systemic immune responses in vaccinated mice. CONCLUSIONS: The results demonstrate that bivalent vaccines targeting α1-giardin and CWP2 can protect mice against the colonization of Giardia trophozoite and block the transformation of cyst in host at the same time, and can be used to prevent Giardia infection and block the transmission of giardiasis.

A Rabbit Model of Acanthamoeba Keratitis That Better Reflects the Natural Human Infection.

Acanthamoeba species are ubiquitous, free-living protozoa that can invade the cornea and result in Acanthamoeba keratitis (AK), a painful progressive sight-threatening corneal disease. Disease progression in current animal models is too rapid to mimic AK in humans accurately. This study provides a novel method for establishing AK in rabbits and compared it with the conventional method with regard to pathogenesis and immune response in humans. The New Zealand white rabbits were randomly divided into two experimental groups (Groups A and B). Rabbits in the Group A (n = 14) received intrastromal injections of 1 × 10(4) /100 µL Acanthamoeba healyi trophozoites (conventional AK model). The Group B animals (n = 14) received microinjections of 1 × 10(4) /10 µL A. healyi trophozoites between the corneal epithelium and Bowman's layer, anterior to the corneal stroma (novel AK model). In addition, two rabbits were left untreated as normal controls. AK in the treated rabbits was evaluated clinically, histopathologically, and immunologically for 35 days. AK was successfully established in both the conventional and novel model groups. Compared with the Group A, AK in the Group B displayed an efficient immune response with less severe pathology. Moreover, the self-limiting but chronic nature of the infection in the Group B was strikingly similar to that of AK in humans. The novel animal model for AK described here more closely simulates the pathogenesis and immune response of Acanthamoeba corneal infection in humans than the animal models currently in use.

[Dynamic changes of serum antibody titer in a rabbit model of Acanthamoeba keratitis].

Ten healthy New Zealand white rabbits were randomly divided into two groups named as experiment group (n=8) and normal control group (n=2). Left eyes were for experiment, right eyes served as control. New Zealand rabbits were each injected by subconjunctival route with hydrocortisone for three days, and then Acanthamoeba keratitis was induced by intrastromal injection of live Acanthamoeba healyi trophozoites and cysts. Eyes in control group were injected with equivalent volume of physiological saline. Corneal lesions of rabbits were recorded every day after injection, etiological diagnosis was carried out by corneal scraping. Blood samples were examined for serum antibody titer by ELISA. Corneas were removed for pathological examination. Corneal scraping and corneal histopathologic examination proved that experiment eyes were infected by Acanthamoeba, and appeared typical manifestations and pathological changes of Acanthamoeba keratitis. Serum antibody titer raised continuously with infection time and reached the highest level (A450 value=2.2358) on the 28th days post-infection, then began to decline and remained higher level than the control until the rabbits were sacrificed. In control group, no significant change in antibody titer had taken place.