Bimetallic nanoplatform for synergistic sonodynamic-calcium overload therapy utilizing self-supplied hydrogen peroxide and relieved hypoxia.

Sonodynamic therapy (SDT) has emerged as a potential alternative to traditional cancer treatments as it offers deep cellular penetration and reduced invasivity. Sonosensitizers generate reactive oxygen species (ROS) under ultrasound activation, focusing the ultrasound energy on malignant sites located deep in tissues and causing cell apoptosis and necrosis. However, due to tumor hypoxia and the limited levels of intracellular endogenous hydrogen peroxide (H2O2 is a fundamental species for supplying oxygen via catalase activity), SDT efficacy is still insufficient. In this study, a bimetallic and multifunctional system (Fe3O4-TAPP@PVP-CaO2) was prepared by using ferrosoferric oxide (Fe3O4) as a carrier loaded with 5,10,15,20-tetrakis(4-aminophenyl), porphyrin (TAPP), that was then coated with polyvinyl pyrrolidone (PVP) and calcium peroxide (CaO2). The CaO2 layer elevated the levels of H2O2 and Ca2+ in the tumor microenvironment when exposed to intracellular acidity, providing essential elements for oxygen generation. Intracellular hypoxia was alleviated via the catalase-like activity of Fe3O4 inducing calcium overload. Under ultrasonic irradiation, SDT generated toxic reactive oxygen species (ROS, singlet oxygen) and activated calcium influx through acoustic cavitation. Meanwhile, calcium overload therapy efficiently induced cell apoptosis at the moment of uncontrollable cellular accumulation of Ca2+. In addition, we modified the PVP on the surface to make it more stable. This study presents a bimetallic nanoplatform that can efficiently induce cancer cell death by synergistic sonodynamic-calcium overload therapy via modulation of O2/ROS/Ca2+ species, indicating its potential for multi-modality cancer therapy.

Key Modulation of ROS and HSP for Effective Therapy Against Hypoxic Tumor with Multifunctional Nanosystem.

Background: Though nanomedicine-based photothermal therapy (PTT) has demonstrated promising prospect in tumor treatment due to its high therapeutic efficiency and controllable range, the overexpression of heat shock proteins (HSPs) during PTT can lead to intracellular thermal resistance and reduce its effectiveness. Reactive oxygen species (ROS), followed by the application of chemodynamic therapy (CDT) and photodynamic therapy (PDT), can eliminate HSPs and overcome thermal resistance. However, the tumor microenvironment, including hypoxia and glutathione (GSH) overexpression, impedes the production of ROS and therapeutic efficacy of CDT and PDT. Therefore, we proposed a multifunctional nanoplatform (HMPB@TCPP-Cu) driving PTT/ PDT/ CDT synergistic therapy for tumor treatment via modulating ROS and HSPs. Methods and Results: In this work, a novel nanoplatform (HMPB@TCPP-Cu) composed of O2/PTT supplier HMPB (hollow mesoporous Prussian blue) and the loaded PDT/CDT agent (TCPP-Cu2+) was prepared. HMPB acts as an photothermal converter, effectively raising the tumor temperature and inducing apoptosis. HMPB is also a potent catalase-like nanozyme, which can catalyze hydrogen peroxide into oxygen and reduce tumor hypoxia, thus elevating the efficiency of ROS production and the effectiveness of PDT with the wing of sonosensitizer-TCPP. The intracellular glutathione(GSH) was depleted by Cu2+ and •OH was generated along with the Cu2+/Cu+ converting and Cu+-mediated Fenton-like reaction. Subsequently, the increased levels of ROS effectively eliminate intratumoral thermal resistance. The HMPB@TCPP-Cu has achieved synergistic PTT/PDT/CDT for hepatoblastoma treatment and significant inhibition of tumor growth was detected both in vitro and in vivo. Conclusion: This study presents a multifunctional nanoplatform that combines photothermal/ chemodynamic/ photodynamic therapy for efficient hepatoblastoma treatment via modulating ROS and HSPs. Collectively, this study provides an appealing strategy in the cleavage of thermal resistance and a novel assistance and enhancement on thermal-related therapies.

Independent wing reductions and losses among stick and leaf insects (Phasmatodea), supported by new Cretaceous fossils in amber.

BACKGROUND: Phasmatodea (stick and leaf insects) play a central role on the debate regarding wing reduction and loss, and its wings are putative reacquisition from secondarily wingless ancestors based solely on extant species. A pivotal taxon in this respect is the species-poor Timematodea, consisting of approximately 21 wingless extant species, which form the sister group of all remaining winged or wingless stick and leaf insects, the Euphasmatodea. RESULTS: Herein, the new fossils of Timematodea from mid-Cretaceous Kachin amber are reported, with winged and wingless species co-occurring. The palaeogeographic distributions of all fossils of Holophasmatodea are summarized, showing their wide paleo-distributions. The phylogenetic analysis based on morphological characters confirms the earliest-diverging lineage of winged Breviala cretacea gen. et sp. nov. in Timematodea, and the possible relationships among all families of Holophasmatodea. These are critical for the reconstruction of patterns of wing evolution in early Phasmatodea. CONCLUSIONS: The new fossils suggest that Timematodea once had wings, at least during the mid-Cretaceous. The palaeogeographic occurrences imply that Timematodea probably have been widely distributed since at least the Jurassic. The phylogenetic analysis with the ancestral-state reconstruction of wings indicates that the common ancestors of Holophasmatodea were winged, the reductions and losses of wings among Timematodea and Euphasmatodea have occurred independently since at least the Cretaceous, and the reduction or loss of the forewing earlier than the hind wings.

Video-Based Neonatal Pain Assessment in Uncontrolled Conditions.

OBJECTIVE: Neonatal pain can have long-term adverse effects on newborns' cognitive and neurological development. Video-based Neonatal Pain Assessment (NPA) method has gained increasing attention due to its performance and practicality. However, existing methods focus on assessment under controlled environments while ignoring real-life disturbances present in uncontrolled conditions. METHODS: We propose a video-based NPA method, which is robust to four real-life disturbances and adaptively highlights keyframes. Our method involves a region-channel-attention module for extracting facial features under the disturbances of facial occlusion and pose variation; a body language analysis module robust to disturbances from body occlusion and movement interference, which utilizes skeleton sequences to represent the neonate's body; and a keyframes-aware convolution to get rid of information located at non-contributing moments. For evaluation, we built an NPA video dataset of 1091 neonates with disturbance annotations. RESULTS: The results show that our method consistently outperforms state-of-the-art methods on the full dataset and nine subsets, where it achieves an accuracy of 91.04% on the full dataset with an accuracy increment of 6.27%. Contributions: We present the problem of video-based NPA under uncontrolled conditions, propose a method robust to four disturbances, and construct a video NPA dataset, thus facilitating the practical applications of NPA.

Evaluation of procedural pain for neonates in a neonatal intensive care unit: a single-centre study.

BACKGROUND: To evaluate the procedural pain experienced by neonates in a neonatal intensive care unit (NICU) setting and determine the corresponding pain grades. METHODS: Two experienced nurses independently used the Neonatal Infant Pain Scale (NIPS) to evaluate the neonatal pain during procedures taking place in the tertiary NICU and two level-two neonatal care units in the Children's Hospital of Zhejiang University School of Medicine. The mean and distribution of NIPS pain scores and the corresponding pain grades of participants when experiencing clinical painful procedures were analysed. RESULTS: A total of 957 neonates exposed to 15 common clinical painful procedures were included in the study. The clinical painful procedures experienced by 957 participants could be divided into three groups: severe pain (NIPS score 5-7: peripheral intravenous cannulation, arterial catheterisation, arterial blood sampling, peripherally inserted central catheter placement and nasopharyngeal suctioning), mild to moderate pain (NIPS score 3-4: finger prick, intramuscular injection, adhesive removal, endotracheal intubation suctioning, heel prick, lumbar puncture and subcutaneous injection) and no pain to mild pain (NIPS score 0-2: gastric tube insertion, enema and intravenous injection). CONCLUSIONS: The neonatal pain response to clinical procedures in NICU had certain pattern and preintervention drug analgesia could be taken for painful procedures with clustered high NIPS pain scores. Meanwhile, full coverage non-drug pain relief measures could be taken for procedures that are with scattered pain scores, and real-time pain evaluation should be provided to determine whether further drug analgesia is required.

Effect of Cinobufacini plus platinum-based chemotherapy regimen on the immune function of patients with non-small-cell lung cancer: A meta-analysis.

Background: Cinobufacini is a Chinese medicinal preparation extracted from the traditional Chinese medicine toad skin and is commonly used clinically as an adjuvant treatment for malignant tumours. Purpose: To systematically evaluate the effects of Cinobufacini combined with a first-line platinum-based chemotherapy regimen in patients with non-small-cell lung cancer (NSCLC), especially in terms of immune function. Materials and methods: Eight electronic databases were searched for randomised controlled trials (RCTs) investigating Cinobufacini in conjunction with platinum-based chemotherapy for NSCLC (stage III-IV) published from 2012 to the present. GRADE Pro GDT was used to assess RCT quality and meta-analysis was performed mainly using Review Manager version 5.4, with the assistance of Stata version 16.0 (StataCorp LLC, College Station, TX, USA), and trial sequential analysis software. Results: A total of 35 studies were included. Meta-analysis revealed that the combination therapy group exhibited a better disease control rate (DCR) [OR = 2.63, 95%CI (2.15, 3.21), P < 0.00001], with a higher one-year [OR = 2.41,95% CI (1.75,3.33), P < 0.00001], and two-year [OR = 2.28, 95% CI (1.56,3.33), P < 0.00001] survival rate, plus lower leukocyte toxicity [OR = 0.40, 95%CI (0.33,0.49), P < 0.00001]. For immune function, the combination of chemotherapy with Cinobufacini effectively increased the proportion of CD3+ [SMD = 1.15, 95% CI (0.89,1.42), P < 0.00001], CD4+ [SMD = 1.60, 95%CI (1.26,1.94), P < 0.00001] and the CD4+/CD8+ ratio [SMD = 2.15, 95% CI (1.45,2.86), P < 0.00001] in peripheral blood. Conclusion: The addition of Cinobufacini to platinum-based chemotherapies for advanced NSCLC significantly improved clinical efficacy, enhanced immune function, and reduced chemotherapeutic toxicity, irrespective of administration and treatment duration.

Pose-invariant and occlusion-robust neonatal facial pain assessment.

Neonatal Facial Pain Assessment (NFPA) is essential to improve neonatal pain management. Pose variation and occlusion, which can significantly alter the facial appearance, are two major and still unstudied barriers to NFPA. We bridge this gap in terms of method and dataset. Techniques to tackle both challenges in other tasks either expect pose/occlusion-invariant deep learning methods or first generate a normal version of the input image before feature extraction, combining these we argue that it is more effective to jointly perform adversarial learning and end-to-end classification for their mutual benefit. To this end, we propose a Pose-invariant Occlusion-robust Pain Assessment (POPA) framework, with two novelties. We incorporate adversarial learning-based disturbance mitigation for end-to-end pain-level classification and propose a novel composite loss function for facial representation learning; compared to the vanilla discriminator that implicitly determines occlusion and pose conditions, we propose a multi-scale discriminator that determines explicitly, while incorporating local discriminators to enhance the discrimination of key regions. For a comprehensive evaluation, we built the first neonatal pain dataset with disturbance annotation involving 1091 neonates and also applied the proposed POPA to the facial expression recognition task. Extensive qualitative and quantitative experiments prove the superiority of the POPA.

Video-Based versus On-Site Neonatal Pain Assessment in Neonatal Intensive Care Units: The Impact of Video-Based Neonatal Pain Assessment in Real-World Scenario on Pain Diagnosis and Its Artificial Intelligence Application.

BACKGROUND: Neonatal pain assessment (NPA) represents a huge global problem of essential importance, as a timely and accurate assessment of neonatal pain is indispensable for implementing pain management. PURPOSE: To investigate the consistency of pain scores derived through video-based NPA (VB-NPA) and on-site NPA (OS-NPA), providing the scientific foundation and feasibility of adopting VB-NPA results in a real-world scenario as the gold standard for neonatal pain in clinical studies and labels for artificial intelligence (AI)-based NPA (AI-NPA) applications. SETTING: A total of 598 neonates were recruited from a pediatric hospital in China. METHODS: This observational study recorded 598 neonates who underwent one of 10 painful procedures, including arterial blood sampling, heel blood sampling, fingertip blood sampling, intravenous injection, subcutaneous injection, peripheral intravenous cannulation, nasopharyngeal suctioning, retention enema, adhesive removal, and wound dressing. Two experienced nurses performed OS-NPA and VB-NPA at a 10-day interval through double-blind scoring using the Neonatal Infant Pain Scale to evaluate the pain level of the neonates. Intra-rater and inter-rater reliability were calculated and analyzed, and a paired samples t-test was used to explore the bias and consistency of the assessors' pain scores derived through OS-NPA and VB-NPA. The impact of different label sources was evaluated using three state-of-the-art AI methods trained with labels given by OS-NPA and VB-NPA, respectively. RESULTS: The intra-rater reliability of the same assessor was 0.976-0.983 across different times, as measured by the intraclass correlation coefficient. The inter-rater reliability was 0.983 for single measures and 0.992 for average measures. No significant differences were observed between the OS-NPA scores and the assessment of an independent VB-NPA assessor. The different label sources only caused a limited accuracy loss of 0.022-0.044 for the three AI methods. CONCLUSION: VB-NPA in a real-world scenario is an effective way to assess neonatal pain due to its high intra-rater and inter-rater reliability compared to OS-NPA and could be used for the labeling of large-scale NPA video databases for clinical studies and AI training.

Cacopsylla fuscicella Sp. Nov. (Hemiptera, Psyllidae), a New Loquat Pest in China.

Psylloidea, commonly known as jumping plant-lice, have a phloem-sucking habit and strong specificity to their host plants. Within the Psyllidae family, the genus Cacopsylla Ossiannilsson, 1970 stands out as being the most diverse, with three species feeding on the genus Eriobotrya Lindl. In this study, a new psyllid species, Cacopsylla fuscicella sp. nov., was described from China. It is a pest of Eriobotrya japonica (Thunb.) Lindl. and has been cultivated as a commercial fruit tree for years. Illustrations of habitus, morphological structures, and its damage to loquat were also provided. The complete mitochondrial genome of C. fuscicella sp. nov. was sequenced and annotated. The phylogenetic tree inferred from the maximum likelihood analyses confirmed C. fuscicella sp. nov. as a member of the genus Cacopsylla. Genetic distances were generated to assess comparative genetic distances within Psyllinae.

Review of Kissing Bugs (Hemiptera: Reduviidae: Triatominae) from China with Descriptions of Two New Species.

Triatominae, the only blood-sucking subfamily in Reduviidae, are the vectors of Chagas disease. The majority of them are distributed in the Americas, while the diversity in China has been underestimated, as only two species have been recorded. Here, we describe two new species from China, Triatoma picta Zhao & Cai sp. nov. and T. atrata Zhao & Cai sp. nov., and provide a redescription of T. sinica Hsiao, 1965, along with remarks on T. rubrofasciata (De Geer, 1773). To facilitate the identification, we include photos, especially of genitalia, as well as a distribution map and a key to Chinese triatomines. We calculated the pairwise genetic distances between 23 Triatoma species, which further supported the validity of these new species. We anticipate that our taxonomic review will be useful for identifying Chinese Triatominae.

Scanning Electron Microscopy of Antennae and Mouthparts of Mezira yunnana Hsiao (Hemiptera: Aradidae): Specialized Microstructures Reflecting Adaptation to Mycetophagy.

Many species of the family Aradidae (also known as flat bugs) feed on fungal mycelia and fruiting bodies. In order to better understand the morphological adaptation to this unique feeding habit, we examined the microstructure of antennae and mouthparts of an aradid species, Mezira yunnana Hsiao, using scanning electron microscope, and documented the fungal feeding process under laboratory conditions. The antennal sensilla include three subtypes of sensilla trichodea, three subtypes of sensilla basiconica, two subtypes of sensilla chaetica, sensilla campaniformia, and sensilla styloconica. The apex of the second segment of flagellum has a large number of various sensilla forming a sensilla cluster. The labial tip is distally constricted, which is rarely observed in other Pentatomomorpha species. The labial sensilla include three subtypes of sensilla trichodea, three subtypes of sensilla basiconica, and a sensilla campaniformia. The tip of the labium has only three pairs of sensilla basiconica III and small comb-shaped cuticular processes. The external surface of the mandibular apex has 8-10 ridge-like central teeth. A series of key morphological structures associated with mycetophagous feeding habit were identified, which will facilitate future studies on adaptive evolution of species in Pentatomomorpha as well as in other heteropteran lineages.

Association between hypertension and the prevalence of liver steatosis and fibrosis.

BACKGROUND: Hypertension (HTN) and non-alcoholic fatty liver disease (NAFLD) frequently coexist and share pathophysiological symptoms. Based on the liver stiffness measurement and controlled attenuation parameter obtained by performing liver transient ultrasound elastography (TUE), we determined the relationship between HTN status and the rates of liver steatosis and fibrosis in this study. METHODS: To perform this cross-sectional study, data were obtained from the National Health and Nutrition Examination Survey for 2017-March 2020 Pre-pandemic cycle. The relationship between HTN and the rates of liver steatosis and fibrosis was analyzed by constructing a multivariate logistic regression model. The VCTE was performed using a FibroScan® system (model 502, V2 Touch), and CAP was measured at ≥ 274 dB/m for liver steatosis, and the LSM result (median, ≥ 8 kPa) confirmed fibrosis. We also conducted subgroup analyses based on the age, sex, ethnicity, and body mass index (BMI) of the patients. RESULTS: In total, 4,705 participants were recruited, including 2,287 participants with HTN and 2,418 without HTN. After adjusting possible confounders, HTN was positively related to the liver steatosis rate (OR = 1.4, 95% CI: 1.1-1.8). Such HTN-associated prevalence was higher among males (OR = 1.6, 95% CI: 1.1-2.2), non-Hispanic African American individuals (OR = 2.1, 95% CI: 1.1-3.7), and participants with BMI ≥ 25 < 30 kg/m2 (OR = 1.7, 95% CI: 1.1-2.5). Additionally, HTN was positively associated with the fibrosis rate (OR = 2.0, 95% CI: 1.3-3.0), especially among females (OR = 2.6, 95% CI: 1.3-5.2), among individuals who were 40-59 years old (OR = 2.1, 95% CI: 1.0-4.3), 60-80 years old (OR = 2.4, 95% CI:1.3-4.6), non-Hispanic Caucasian (OR = 2.9, 95% CI: 1.5-5.6), among those with BMI ≥ 25 < 30 kg/m2 (OR = 3.0, 95% CI: 1.1-8.2), and those with BMI ≥ 30 kg/m2 (OR = 2.1, 95% CI: 1.4-3.2). CONCLUSION: The results of this study revealed that HTN status was associated with higher rates of liver steatosis and fibrosis, particularly in subjects with BMI ≥ 25 kg/m2. The ethnicity of the participants also had an impact on the relationship.

Artificial Intelligence Based Pain Assessment Technology in Clinical Application of Real-World Neonatal Blood Sampling.

Background: Accurate neonatal pain assessment (NPA) is the key to neonatal pain management, yet it is a challenging task for medical staff. This study aimed to analyze the clinical practicability of the artificial intelligence based NPA (AI-NPA) tool for real-world blood sampling. Method: We performed a prospective study to analyze the consistency of the NPA results given by a self-developed automated NPA system and nurses' on-site NPAs (OS-NPAs) for 232 newborns during blood sampling in neonatal wards, where the neonatal infant pain scale (NIPS) was used for evaluation. Spearman correlation analysis and the degree of agreement of the pain score and pain grade derived by the NIPS were applied for statistical analysis. Results: Taking the OS-NPA results as the gold standard, the accuracies of the NIPS pain score and pain grade given by the automated NPA system were 88.79% and 95.25%, with kappa values of 0.92 and 0.90 (p < 0.001), respectively. Conclusion: The results of the automated NPA system for real-world neonatal blood sampling are highly consistent with the results of the OS-NPA. Considering the great advantages of automated NPA systems in repeatability, efficiency, and cost, it is worth popularizing the AI technique in NPA for precise and efficient neonatal pain management.

On-tissue derivatization for isomer-specific mass spectrometry imaging and relative quantification of monosaccharides in biological tissues.

Spatially-resolved profiling of tissue monosaccharides not only gives an insight into the spatial heterogeneity of monosaccharides, but also helps to decipher the possible roles of monosaccharides in biological processes. Here, we develop an on-tissue derivatization method, coupled with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) to image and quantify the aldose and ketose isomers of monosaccharide in biological tissues. A new derivatization reagent, 1-naphthaleneacethydrazide (NAH) was synthesized for the on-tissue derivatization of monosaccharides, and it significantly enhanced the imaging sensitivity of monosaccharides. Moreover, the NAH-derivatized aldose and ketose can generate isomer-specific diagnostic ions during MALDI-MS/MS analysis, and thus paves way for the isomer-specific MS imaging of aldose and ketose monosaccharides. On this basis, we further constructed a quantitative MALDI-MS imaging model based on isomer-specific diagnostic ions, and calculated the expression contents of aldose and ketose monosaccharide isomers in different tissue regions of carrot section. We expect that the development of this method should provide more precise view on the spatial distributions and contents of different monosaccharides in heterogeneous biological tissues.

Robust Heartbeat Classification for Wearable Single-Lead ECG via Extreme Gradient Boosting.

Wearable electrocardiogram (ECG) monitoring devices have enabled everyday ECG collection in our daily lives. However, the condition of ECG signal acquisition using wearable devices varies and wearable ECG signals could be interfered with by severe noises, resulting in great challenges of computer-aided automated ECG analysis, especially for single-lead ECG signals without spare channels as references. There remains room for improvement of the beat-level single-lead ECG diagnosis regarding accuracy and efficiency. In this paper, we propose new morphological features of heartbeats for an extreme gradient boosting-based beat-level ECG analysis method to carry out the five-class heartbeat classification according to the Association for the Advancement of Medical Instrumentation standard. The MIT-BIH Arrhythmia Database (MITDB) and a self-collected wearable single-lead ECG dataset are used for performance evaluation in the static and wearable ECG monitoring conditions, respectively. The results show that our method outperforms other state-of-the-art models with an accuracy of 99.14% on the MITDB and maintains robustness with an accuracy of 98.68% in the wearable single-lead ECG analysis.

Rhodnius micki, a new species of Triatominae (Hemiptera, Reduviidae) from Bolivia.

Rhodnius Stål, 1859 is the second largest genus of Triatominae after Triatoma Laporte, 1832, and includes several important Chagas vectors. Genitalia in Reduviidae are frequently used for species identification, but the current use of terminology for it is inconsistent in Triatominae. Here, Rhodnius micki sp. nov., is described from Bolivia and considered as belonging to the pictipes group based on its morphological characters and distribution. Detailed documentation of the genitalia of Rhodnius micki sp. nov. is provided with emphasis on its everted phallus, especially the endosomal sclerites, which are potentially useful as species-level diagnostic features in Rhodnius. To further verify the validity of this species, the head shapes and wing venation patterns of five species in Rhodnius are compared with morphometric analysis. After reviewing taxonomic and comparative morphology papers of assassin bugs, a vocabulary with a terminology of morphological characters, especially of external male genitalic characters, is assembled with the preferred terms and the synonyms listed. Establishing a consistent terminological framework will greatly facilitate future research on the homology of these structures across Triatominae and will ultimately contribute to our understanding of the evolution of these groups.

Immunogenicity Assessment of Different Segments and Domains of Group a Streptococcal C5a Peptidase and Their Application Potential as Carrier Protein for Glycoconjugate Vaccine Development.

Group A streptococcal C5a peptidase (ScpA) is a highly conserved surface virulence factor present on group A streptococcus (GAS) cell surfaces. It has attracted much more attention as a promising antigenic target for GAS vaccine development due to its high antigenicity to stimulate specific and immunoprotective antibodies. In this study, a series of segments of ScpA were rationally designed according to the functional domains described in its crystal structure, efficiently prepared and immunologically evaluated so as to assess their potential as antigens for the development of subunit vaccines. Immunological studies revealed that Fn, Fn2, and rsScpA193 proteins were promising antigen candidates worthy for further exploration. In addition, the potential of Fn and Fn2 as carrier proteins to formulate effective glycoconjugate vaccine was also investigated.

Group A Streptococcus Cell Wall Oligosaccharide-Streptococcal C5a Peptidase Conjugates as Effective Antibacterial Vaccines.

Group A streptococcus (GAS) is one of the common Gram-positive pathogenic bacteria accounting for a variety of infectious diseases. Currently, there is no commercial vaccine for GAS. To develop efficient GAS vaccines, synthetic tri-, hexa-, and nonasaccharides of a conserved group A carbohydrate (GAC) were conjugated with an inactive mutant of group A streptococcal C5a peptidase (ScpA), ScpA193, to create bivalent conjugate vaccines, which were compared with the corresponding CRM197 and TT conjugates. Systematic evaluations of these semisynthetic conjugates demonstrated that they could induce robust and comparable T-cell-dependent immune responses in mice. It was further disclosed that antibodies provoked by the ScpA193 conjugates, especially that of hexa- and nonasaccharides, could recognize and bind to GAS cells and mediate GAS opsonophagocytosis in vitro. In vivo evaluations of the hexa- and nonasaccharide-ScpA193 conjugates using a mouse model revealed that immunizing mice with especially the latter conjugate could effectively protect the animals from GAS challenges and GAS-induced pulmonary damage and significantly increase animal survival. Further in vitro studies suggested that the two ScpA193 conjugates could function through activating CD4+ T cells and promoting helper T cells (Th) to differentiate into antigen-specific Th1 and Th2 cells. In conclusion, the nonasaccharide-ScpA193 conjugate was identified as a particularly promising GAS vaccine candidate that is worthy of further investigation and development.

Mitochondrial genomes of three kissing bugs (Reduviidae: Triatominae) and their phylogenetic implications.

The reduviid subfamily Triatominae, also called kissing bugs, are vectors of Chagas disease, which is one of the most seriously neglected tropical parasitic diseases. Only three complete mitochondrial genomes of kissing bugs from the genus Triatoma have been sequenced to date. To better understand the diversity of mitochondrial genomes and the evolution of kissing bugs, mitochondrial genomes of three kissing bugs, Triatoma migrans, Panstrongylus rufotuberculatus, and Rhodnius pictipes, were sequenced using next-generation sequencing and a comparative mitochondrial genomic analysis of three genera and two tribes in Triatominae was conducted. Kissing bug mitochondrial genomes shared a similar pattern of nucleotide composition, gene order, and structure of control region. The comparison among orthologous protein-coding genes indicated that different genes had different rates of molecular evolution and six genes (ND1, ND2, ND4L, ND5, ND6, and ATP8) had higher evolutionary rates than other protein-coding genes. Phylogenetic analyses inferred from mitochondrial genome sequences supported a sister relationship between Triatominae and Stenopodainae, and the genus Triatoma was paraphyletic. The present study revealed the high conservation of the mitochondrial genome organization of kissing bugs and highlighted the utility of mitochondrial genomes in the phylogenetic study of Triatominae.

The mitochondrial genome of the ambush bug Carcinochelis bannaensis (Hemiptera: Reduviidae).

The complete mitochondrial genome (mitogenome) of the ambush bug, Carcinochelis bannaensis, was determined in this study. The sequenced mitogenome is a typical circular DNA molecule of 15,335 bp, containing 13 protein-coding genes, two rRNA genes, 22 tRNA genes and a putative control region. All protein-coding genes initiate with ATN codons and terminate with TAA codons except for COII,COIII and ND5 use a single T residue as the stop codon. All tRNAs have the clover-leaf structure except for the tRNASer(AGN) and the length of them range from 61 to 71 bp. The control region is 797 bp long with an A + T content of 66.3%. The phylogenetic analysis result supports the monophyly of Phymatinae.