Effects of intrauterine infection in different periods on the placenta and endometrial blood vessel formation of pregnant mice and the growth and development of fetal rats

Objective: To investigate the effects of intrauterine infection in different periods on the placenta and endometrial blood vessel formation of pregnant rats and the growth and development of fetal rats.Methods: According to the random number table method, 32 pregnant rats were divided into the early infection group, the mid-term infection group, the late infection group and the control group, with 8 rats in each group. On the 3rd, 9th and 15th day of pregnancy, lipopolysaccharide was injected intraperitoneally to construct intrauterine infection models. The pregnant rats in the control group were intraperitoneally injected with the same dose of 0.9% sodium chloride solution. On the 18th day of pregnancy, the inflammatory factors [interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α)], the blood vessel density of placenta and endometrium in the placental tissues of pregnant rats, dead fetus + absorbed fetus, the inflammatory factors IL-6, TNF-α and oxidation reaction indicators [malondialdehyde (MDA) and myeloperoxidase (MPO)] in the fetal rat lung and brain tissues were detected.Results: The changing trend of IL-6 and TNF-α levels in the placental tissues of pregnant rats with intrauterine infection in different periods was: the control group < the late infection group < the mid-term infection group < the early infection group, the differences were statistically significant (p <.05). The changing trend of fetal rat weight, placental weight and placental coefficient in the intrauterine infection groups in different periods was: the control group > the late infection group > the mid-term infection group > the early infection group, the differences were statistically significant (p < .05). The blood vessel density of placenta and endometrium, the mean number of fetuses, brain coefficient and lung coefficient in the late infection group were significantly increased in comparison with the early infection group and the mid-term infection group. The total number and the ratio of dead fetus + absorbed fetus, the levels of IL-6, TNF-α, MDA and MPO in brain and lung tissues were significantly reduced, and the differences were statistically significant (p < .05). The blood vessel density of placenta and endometrium, brain coefficient and lung coefficient of pregnant rats in the mid-term infection group were significantly increased in comparison with the early infection group, and the differences were statistically significant (p < .05). There was no statistically significant difference in the other indicators between the two groups (p > .05).Conclusions: Intrauterine infection in different periods can inhibit placental and endometrial angiogenesis, and affect the survival rate of fetal rats and the growth and development of brain and lung. The reason may be related to the aggravation of fetal inflammatory responses and oxidative stress. The earlier the intrauterine infection occurs, the severer the adverse effects on the fetal rats will be.

Botrytis cinerea BcPTP1 is a late infection phase, cysteine rich protein cytotoxic effector

Botrytis cinerea is a broad-host-range necrotrophic phytopathogen responsible for serious crops diseases. To facilitate infection, B. cinerea secretes a large number of effectors that induce plant cell death. In screening secretome data of B. cinerea during infection stage, we identified a phytotoxic protein (BcPTP1) that can also induce immune resistance in plants. BcPTP1 is a small (90 aa), cysteine rich protein without any known domains. Transiently expression of BcPTP1 in leaves caused chlorosis that intensifies with time and eventually lead to cell death. Point mutations in eight of the 10 cysteine residues of BcPTP1 abolished the toxic effect, however residual toxic activity remained after heating the peptide, suggesting contribution of unknown epitopes to protein phytotoxic effect. The transcript level of the bcptp1 gene was low during the first 36 h after inoculation and increased sharply upon transition to the late infection stage, suggesting a role of BcPTP1 in lesion spreading. While statistically insignificant, deletion of the bcptp1 gene led to slightly smaller lesions on bean leaves. Further analyses indicated that BcPTP1 is internalized into plant cells after secreting into the apoplast and its phytotoxic effect is negatively regulated by the receptor-like kinases BAK1 and SOBIR1. Collectively, our findings show that BcPTP1 is a virulence factor that toxifies the host cells and facilitates lesion spreading during the late infection stage.

Dual species dynamic transcripts reveal the interaction mechanisms between Chrysanthemum morifolium and Alternaria alternata

Background Chrysanthemum (Chrysanthemum morifolium) black spot disease caused by Alternaria alternata is one of the plant’s most destructive diseases. Dual RNA-seq was performed to simultaneously assess their transcriptomes to analyze the potential interaction mechanism between the two species, i.e., host and pathogen. Results C. morifolium and A. alternata were subjected to dual RNA-seq at 1, 12, and 24 h after inoculation, and differential expression genes (DEGs) in both species were identified. This analysis confirmed 153,532 DEGs in chrysanthemum and 14,932 DEGs in A. alternata, which were involved in plant-fungal interactions and phytohormone signaling. Fungal DEGs such as toxin synthesis related enzyme and cell wall degrading enzyme genes played important roles during chrysanthemum infection. Moreover, a series of key genes highly correlated with the early, middle, or late infection stage were identified, together with the regulatory network of key genes annotated in the Plant Resistance Genes database (PRGdb) or Pathogen-Host Interactions database (PHI-base). Highly correlated genes were identified at the late infection stage, expanding our understanding of the interplay between C. morifolium and A. alternata. Additionally, six DEGs each from chrysanthemum and A. alternata were selected for quantitative real-time PCR (qRT-PCR) assays to validate the RNA-seq output. Conclusions Collectively, data obtained in this study enriches the resources available for research into the interactions that exist between chrysanthemum and A. alternata, thereby providing a theoretical basis for the development of new chrysanthemum cultivars with resistance to pathogen.

Autophagy Receptor Protein Tax1-Binding Protein 1/TRAF6-Binding Protein Is a Cellular Substrate of Enteroviral Proteinase

Enteroviruses (EVs) usurp the host autophagy pathway for pro-viral functions; however, the consequence of EV-induced diversion of autophagy on organelle quality control is poorly defined. Using coxsackievirus B3 (CVB3) as a model EV, we explored the interplay between EV infection and selective autophagy receptors, i.e., Tax1-binding protein 1/TRAF6-binding protein (T6BP), optineurin (OPTN), and nuclear dot 10 protein 52 (NDP52), known to be involved in regulating the clearance of damaged mitochondria, a process termed as mitophagy. Following CVB3 infection, we showed significant perturbations of the mitochondrial network coincident with degradation of the autophagy receptor protein T6BP, similar phenomenon to what we previously observed on NDP52. Notably, protein levels of OPTN are not altered during early infection and slightly reduced upon late infection. Cell culture studies revealed that T6BP degradation occurs independent of the function of host caspases and viral proteinase 3C, but requires the proteolytic activity of viral proteinase 2A. Further investigation identified the cleavage site on T6BP after the amino acid 621 that separates the C-terminal ubiquitin-binding domain from the other functional domains at the N-terminus. Genetic silencing of T6BP and OPTN results in the attenuation of CVB3 replication, suggesting a pro-viral activity for these two proteins. Finally, functional assessment of cleaved fragments from NDP52 and T6BP revealed abnormal binding affinity and impaired capacity to be recruited to depolarized mitochondria. Collectively, these results suggest that CVB3 targets autophagy receptors to impair selective autophagy.

Starch Catabolism Revealed during Secondary Metabolite Released Under Vascular Streak Dieback Infections in Cocoa (Theobroma cacao L)

This study aimed to study the profile of starch content in cocoa leaf and phytoalexin production based on GC-MS analysis at several stages of VSD pathogen infection. Research was conducted on January – October 2015 at Kaliwining Experimental Field, Indonesian Coffee and Cocoa Research Institute, Jember, East Java. The research was designed based on a Completely Randomized Block Design with two factors with three replications. The first factor was clone, i.e. the resistant clone (Scavina 6) and susceptible (TSH 858) to VSD infection. The second factor was the level of O. theobromae infection, i.e. pre-infection, early infection, and late infection. Starch catabolism revealed during Vascular Streak Dieback infections in Cacao. Starch content in Sca 6 (resistant clone) in late infection decreased 24,33 % than healthy condition (no infection), however, starch content in TSH 858 (succeptible clone) in late infection decreased only 9,63 % than healthy condition (no infection). This indicated that starch catabolism rate on resistant clone was higher than susceptible clone. Some secondary metabolites releases under Vascular Streak Dieback i.e. I-limonene, eugenol and coumaran. Scavina 6 (resistant clone) had higher concentration of eugenol and coumaran than TSH 858 (susceptible clone). I-limonene compound, TSH 858 (Susceptible clone) had higher concentration than Scavina 6. I-Limonene concentration increased in lined with the severity of pathogen infection. There were an negative correlation between starch content with contentration of I-limonene (R= - 0,74), concentration of Eugenol (R= - 0,44), and contentration of Coumaran.

Anti-phospholipids antibodies and immune complexes in COVID-19 patients: a putative role in disease course for anti-annexin-V antibodies

Introduction Besides distinctive respiratory and digestive hallmarks, COVID-19 has been recently associated with a high prevalence of pro-inflammatory and hypercoagulable states known as “COVID-19 Associated Coagulopathy” (CAC), corresponding to a worsening in patients’ conditions, whose causes are still to be elucidated. A link between anti-phospholipid antibodies (aPLs) and viral infections has long been suggested. APLs are assessed for anti-phospholipid syndrome (APS) diagnosis, characterized by thrombocytopenia, thrombosis, and coagulopathy. Furthermore, circulating immune complexes (CICs), arisen upon inflammatory responses and related immune dysregulation, can lead to endothelial cell damage and thrombotic complications. Method We performed an extended panel including IgG/IgM anti-cardiolipin, IgG/IgM anti-β2-glycoprotein-1, coupled with IgG/IgM anti-prothrombin, IgG/IgM anti-annexin-V on two COVID-19 patient groups (early and late infection time), and a negative control group. IgG CIC analysis followed to evaluate inflammatory status, through a possible complement system activation. Results Our results showed low positive case percentage in IgG/IgM anti-cardiolipin and IgG/IgM anti-β2-glycoprotein-1 assays (4.54%, 6.25%, and 4.55%; in early infection group, late infection group, and control group, respectively); few positive cases in IgG/IgM anti-prothrombin and IgG/IgM anti-annexin-V immunoassays; and no IgG CIC positivity in any patient. Conclusions In conclusion, our data show a low aPL prevalence, likely excluding an involvement in the pathogenesis of CAC. Interestingly, IgG/IgM anti-prothrombin and anti-annexin-V positive cases, detected in late infection group, suggest that aPLs could temporarily increase or could trigger a “COVID-19-induced-APS-like-syndrome” in predisposed patients. Key Points•To our knowledge, anti-prothrombin (aPT) antibodies, anti-annexin-V antibodies and CICs in COVID-19 patients have not been reported in the scientific literature.•Lack of uniformity and the low percentage of aCL/aβ2GP1 positivity preclude a putative role in CAC pathogenesis.•IgG/IgM anti-prothrombin and IgG/IgM anti-annexin-V data show that distribution of positive case number increases in late infection patients, significantly in anti-annexin-V results, suggesting a possible role for these anti-phospholipid antibodies in disease course.•aPLs can arise transiently in some patients with critical illness and SARS-CoV-2 infection (disappearing in a few weeks), as well as in other genetically predisposed patients; they could trigger a “COVID-19-induced-APS-like-syndrome”.

Safety and Efficacy of Chloroquine/Hydroxychloroquine in SARS-CoV-2 Infection

This work summarizes the available evidence of the use of chloroquine/hydroxychloroquine (CQ/HCQ) in SARS-CoV-2 infection. Most of the published works indicate CQ/HCQ is likely effective against SARS-CoV-2 infection, almost 100% in prophylaxis and mild-medium severity cases and 60% in late infection cases. The percentage of positive works is larger if those works conducted under a probable conflict of interest are excluded from the list. Despite this overwhelming evidence from independent studies, the use of CQ/HCQ is currently limited or prevented in many western countries, based on a very singular examination of the science. The case of a work published in late May 2020, despite being openly defective and then retracted, prompted the World Health Organization (WHO) to ban the use of CQ/HCQ. This position has not yet rectified, thanks to the results of the not less questionable RECOVERY trial, where very sick patients were administered more than double the dose, over more than double the time, recommended for asymptomatic patients in current protocols of other countries, where CQ/HCQ are used for asymptomatic and mild but not severe pneumonia critically ill patients. While the case fatality rate does not depend only on therapies, it is finally shown based on the number of cases and fatalities per million and the case fatality rate as the western countries enforcing the ban on CQ/HCQ did not perform better, but much worse, than other countries, also because of therapies.

Dynamics of Schistosoma Mansoni Development in The Intermediate Host, Biomphalaria Glabrata, A Freshwater Snail

Background Schistosomiasis is one of the most significant and prevalent waterborne parasitic diseases. Even though many studies have been reported about schistosomiasis, the dynamics of schistosome in intermediate host snails is little known. In the present study, the dynamics of Schistosoma larvae in infected snails was histologically investigated. Methods To examine the localization of Schistosoma mansoni (S. mansoni) parasites in the snails, Biomphalaria glabrata snails infected with miracidia were harvested and examined by stereoscopic observation. Then, frozen sections were prepared and stained with H&E. Furthermore, immunohistochemical detection of parasites was performed using anti-S. mansoni antibody, and their localization in the snails was analyzed. Results Snails infected with S. mansoni miracidia were harvested at 10 and 56 days post-infection (DPI) and analyzed. In the stereoscopic observations, white spots were observed at 56 DPI, while no spots were observed at 10 DPI. However, histological investigations visualized the larvae specifically in the head-foot area of the snail at 10 DPI. Further, it was observed that the larvae relocated to the hepatopancreas and ovotestis areas at 56 DPI. Conclusions The present study revealed the dynamics of Schistosoma larvae in intermediate snails, shown as the differential localization of S. mansoni larvae at early and late infection stages.

Differential Apoptotic Responses of Hemocyte Subpopulations to White Spot Syndrome Virus Infection in Fenneropenaeus chinensis

The apoptosis of hemocytes plays an essential function in shrimp immune defense against pathogen invasions. In order to further elucidate the differential apoptotic responses of the granulocytes and the hyalinocytes in Fenneropenaeus chinensis post WSSV infection, the characteristics of apoptotic dynamics and viral proliferation in total hemocytes and hemocyte subpopulations were respectively investigated in the present work. The results showed that the apoptotic rate of hemocytes changed significantly, and the apoptosis-related genes also showed significantly differential expression responses during WSSV infection. Interestingly, we found that the apoptotic rate of virus-negative hemocytes was significantly higher than that of virus-positive hemocytes in the early stage of WSSV infection, while it was significantly lower than that of virus-positive cells in the middle and late infection stages. The difference of apoptosis between virus-positive and virus-negative hemocytes seems to be an important way for the WSSV to destroy the host’s immune system and facilitate the virus spread at different infection stages. It was further found that the apoptosis rate of granulocytes was always significantly higher than that of hyalinocytes during WSSV infection, indicating that granulocytes have a stronger apoptotic response to WSSV infection. Moreover, a higher viral load was detected in granulocytes, and the density of granulocytes decreased more rapidly post WSSV infection, indicating that the granulocytes are more susceptible and vulnerable to WSSV infection compared with the hyalinocytes. These results collectively demonstrated that the apoptotic response in shrimp hemocytes was significantly influenced by the WSSV infection, and the differential apoptotic response of granulocytes and hyalinocytes to WSSV indicated the differences of antiviral mechanisms between the two hemocyte subpopulations.

Dual species dynamic transcripts reveal the interaction mechanisms between Chrysanthemum morifolium and Alternaria alternata

Background: Chrysanthemum (C. morifolium) black spot disease caused by Alternaria alternata is one of the plant’s most destructive diseases. Dual RNA-seq was performed to simultaneously assess their transcriptomes to analyze the potential interaction mechanism between the two species, i.e., host and pathogen. Results: C. morifolium and A. alternata were subjected to dual RNA-seq at 1, 12, and 24 hours after inoculation, and differential expression genes (DEGs) in both species were identified. This analysis confirmed 153,532 DEGs in chrysanthemum and 14,932 DEGs in A. alternata, that were involved in plant-fungal interactions and phytohormone signaling. Fungal DEGs such as toxin synthesis related enzyme and cell wall degrading enzyme genes played important roles during chrysanthemum infecton. Moreover, a series of key genes highly correlated with the early, middle, or late infection stage was identified, together with the regulatory network of key genes annotated in PRG or PPI databases. Highly correlated genes were identified at the late infection stage, expanding our understanding of the interplay between C. morifolium and A. alternata. Additionally, six DEGs each from chrysanthemum and A. alternata were selected for qRT-PCR assays to validate the RNA-seq output. Conclusions: Collectively, data obtained in this study enriches the resources available for research into the interactions that exist between chrysanthemum and A. alternata, thereby providing a theoretical basis for the development of new chrysanthemum cultivars with resistance to pathogen.