TLR9 and STING agonists synergistically induce innate and adaptive type‐II IFN

Agonists for TLR9 and Stimulator of IFN Gene (STING) act as vaccine adjuvants that induce type‐1 immune responses. However, currently available CpG oligodeoxynucleotide (ODN) (K‐type) induces IFNs only weakly and STING ligands rather induce type‐2 immune responses, limiting their potential therapeutic applications. Here, we show a potent synergism between TLR9 and STING agonists. Together, they make an effective type‐1 adjuvant and an anticancer agent. The synergistic effect between CpG ODN (K3) and STING‐ligand cyclic GMP–AMP (cGAMP), culminating in NK cell IFN‐γ (type‐II IFN) production, is due to the concurrent effects of IL‐12 and type‐I IFNs, which are differentially regulated by IRF3/7, STING, and MyD88. The combination of CpG ODN with cGAMP is a potent type‐1 adjuvant, capable of inducing strong T_h1‐type responses, as demonstrated by enhanced antigen‐specific IgG2c and IFN‐γ production, as well as cytotoxic CD8⁺ T‐cell responses. In our murine tumor models, intratumoral injection of CpG ODN and cGAMP together reduced tumor size significantly compared with the singular treatments, acting as an antigen‐free anticancer agent. Thus, the combination of CpG ODN and a STING ligand may offer therapeutic application as a potent type‐II IFN inducer.     

Early to middle gestational exposure to diethylstilbestrol impairs the development of labyrinth zone in mouse placenta

This study was performed to clarify the involvement of impaired labyrinth zone (LZ) of the placenta in the developmental toxicity of diethylstilbestrol (DES). DES at 10 μg/kg per day was administered orally to mice on days 4 through 8 of gestation. Histological observation of the LZ and determination of blood glucose levels in dam and fetus were performed on day 13. A high frequency of embryonic death was observed in the DES group. DES induced the underdevelopment of the plexus vasculosus, extensive maternal blood space and the decreased expression of glucose transporters in the LZ, and a reduction of the glucose level in embryos. These findings suggest that impaired LZ development may be related to the embryolethality of DES.     

Jerky spontaneous movements at term age in preterm infants who later developed cerebral palsy

Background

Assessment of spontaneous movements in infants has been a powerful predictor of cerebral palsy (CP). Recent advancements on computer-based video analysis can provide detailed information about the properties of spontaneous movements.

Aims

The aim of this study was to investigate the relationship between spontaneous movements of the 4 limbs at term age and the development of CP at 3 years of age by using a computer-based video analysis system.

Study design and subjects

We analyzed video recordings of spontaneous movements at 36–44 weeks postmenstrual age (PMA) for 145 preterm infants who were born preterm (22–36 weeks PMA with birthweights of 460–1498 g). Sixteen of the infants developed CP by 3 years of age, while 129 developed normally. We compared 6 movement indices calculated from 2-dimensional trajectories of all limbs between the 2 groups.

Results

We found that the indices of jerkiness were higher in the CP group than in the normal group (p < 0.1 for arms and p < 0.01 for legs). No decline was observed in the average velocity and number of movement units in the CP group compared with to the normal group.

Conclusions

Jerkiness of spontaneous movements at term age provides additional information for predicting CP in infants born preterm.     

Genetic Diversity and Dynamic Distribution of Mycobacterium tuberculosis Isolates Causing Pulmonary and Extrapulmonary Tuberculosis in Thailand

This study examined the genetic diversity and dynamicity of circulating Mycobacterium tuberculosis strains in Thailand using nearly neutral molecular markers. The single nucleotide polymorphism (SNP)-based genotypes of 1,414 culture-positive M. tuberculosis isolates from 1,282 pulmonary tuberculosis (PTB) and 132 extrapulmonary TB (EPTB) patients collected from 1995 to 2011 were characterized. Among the eight SNP cluster groups (SCG), SCG2 (44.1%), which included the Beijing (BJ) genotype, and SCG1 (39.4%), an East African Indian genotype, were dominant. Comparisons between the genotypes of M. tuberculosis isolates causing PTB and EPTB in HIV-negative cases revealed similar prevalence trends although genetic diversity was higher in the PTB patients. The identification of 10 reported sequence types (STs) and three novel STs was hypothesized to indicate preferential expansion of the SCG2 genotype, especially the modern BJ ST10 (15.6%) and ancestral BJ ST19 (13.1%). An association between SCG2 and SCG1 genotypes and particular patient age groups implies the existence of different genetic advantages among the bacterial populations. The results revealed that increasing numbers of young patients were infected with M. tuberculosis SCGs 2 and 5, which contrasts with the reduction of the SCG1 genotype. Our results indicate the selection and dissemination of potent M. tuberculosis genotypes in this population. The determination of heterogeneity and dynamic population changes of circulating M. tuberculosis strains in countries using the Mycobacterium bovis BCG (bacillus Calmette-Guérin) vaccine are beneficial for vaccine development and control strategies.     

The zoonotic potential of avian influenza viruses isolated from wild waterfowl in Zambia

Whilst remarkable progress in elucidating the mechanisms governing interspecies transmission and pathogenicity of highly pathogenic avian influenza viruses (AIVs) has been made, similar studies focusing on low-pathogenic AIVs isolated from the wild waterfowl reservoir are limited. We previously reported that two AIV strains (subtypes H6N2 and H3N8) isolated from wild waterfowl in Zambia harbored some amino acid residues preferentially associated with human influenza virus proteins (so-called human signatures) and replicated better in the lungs of infected mice and caused more morbidity than a strain lacking such residues. To further substantiate these observations, we infected chickens and mice intranasally with AIV strains of various subtypes (H3N6, H3N8, H4N6, H6N2, H9N1 and H11N9) isolated from wild waterfowl in Zambia. Although some strains induced seroconversion, all of the tested strains replicated poorly and were nonpathogenic for chickens. In contrast, most of the strains having human signatures replicated well in the lungs of mice, and one of these strains caused severe illness in mice and induced lung injury that was characterized by a severe accumulation of polymorphonuclear leukocytes. These results suggest that some strains tested in this study may have the potential to infect mammalian hosts directly without adaptation, which might possibly be associated with the possession of human signature residues. Close monitoring and evaluation of host-associated signatures may help to elucidate the prevalence and emergence of AIVs with potential for causing zoonotic infections.     

Long-term results of balloon-occluded retrograde transvenous obliteration for gastric variceal bleeding and risky gastric varices: a 10-year experience

Background and Aim: Balloon‐occluded retrograde transvenous obliteration (B‐RTO) is a new alternative treatment for gastric varices (GVx), but the long‐term efficacy is not known. We investigated the long‐term effects of B‐RTO on rebleeding, prevention of first bleeding, mortality and occurrence of risky esophageal varices (EVx). Methods: B‐RTO was performed in 68 cirrhotic patients with GVx. Twenty patients had recent bleeding, transiently treated by endoscopic Histoacryl injection or balloon tamponade. Forty‐eight patients had varices likely to bleed, but no bleeding. After B‐RTO, the recurrent bleeding, occurrence of EVx and mortality over the long‐term were evaluated. Results: B‐RTO was successfully performed in 63 of 68 patients (92.6%). Varices eradication was confirmed by endoscopy in 61 of 63 patients (96.6%). During follow up, GVx bleeding occurred in two patients (3.2%). The 8‐year cumulative rebleeding rates of patients with bleeding and risky GVx were 14% and 0%, respectively. Risky EVx occurred in 10 patients (17%) and the cumulative occurrence rate was 22% in 8 years. The cumulative occurrence rate of risky EVx was higher in GVx with EVx (GOV2‐GVx) compared to GVx without EVx (IGV1, P < 0.05). No ectopic variceal bleeding occurred. No patients died from variceal bleeding. Hepatocellular carcinoma was the only significant prognostic factor (P < 0.05). Conclusion: B‐RTO is beneficial over the long‐term, despite worsening EVx in some patients, because of excellent treatment efficacy and improved mortality. We believe that B‐RTO can become a first‐choice radical treatment following hemostasis for gastric variceal bleeding and prophylactic treatment for risky GVx.     

Cilia play a role in breaking left–right symmetry of the sea urchin embryo

Left–right asymmetry of bilaterian animals is established during early development. In mice, frogs and fishes, the ciliated left–right organizer plays an essential role in establishing bilateral asymmetry, and leftward flow of extracellular fluid generated by ciliary motion results in Nodal activity on the left side. However, H⁺/K⁺‐ATPase activity is also involved in the determination of left–right asymmetry in a variety of animals, and it has been thought to be an ancestral mechanism in deuterostomes. In sea urchin, the determination of the left–right asymmetry based on H⁺/K⁺‐ATPase activity was already clarified, but it remains to be uncovered whether ciliary motion is involved in the left–right asymmetry of the embryo. Here, we show evidence that ciliary motion is involved in the establishment of left–right asymmetry of sea urchin embryo. Furthermore, we show that the initial cilia generated on small micromeres during the early stage of embryogenesis may be involved in this process. These results suggest that the cilia‐mediated mechanism for the determination of left–right asymmetry may be acquired at the base of the deuterostomes.