Melatonin alleviates lung injury in H1N1-infected mice by mast cell inactivation and cytokine storm suppression.

Influenza A virus (IAV) H1N1 infection is a constant threat to human health and it remains so due to the lack of an effective treatment. Since melatonin is a potent antioxidant and anti-inflammatory molecule with anti-viral action, in the present study we used melatonin to protect against H1N1 infection under in vitro and in vivo conditions. The death rate of the H1N1-infected mice was negatively associated with the nose and lung tissue local melatonin levels but not with serum melatonin concentrations. The H1N1-infected AANAT-/- melatonin-deficient mice had a significantly higher death rate than that of the WT mice and melatonin administration significantly reduced the death rate. All evidence confirmed the protective effects of melatonin against H1N1 infection. Further study identified that the mast cells were the primary targets of melatonin action, i.e., melatonin suppresses the mast cell activation caused by H1N1 infection. The molecular mechanisms involved melatonin down-regulation of gene expression for the HIF-1 pathway and inhibition of proinflammatory cytokine release from mast cells; this resulted in a reduction in the migration and activation of the macrophages and neutrophils in the lung tissue. This pathway was mediated by melatonin receptor 2 (MT2) since the MT2 specific antagonist 4P-PDOT significantly blocked the effects of melatonin on mast cell activation. Via targeting mast cells, melatonin suppressed apoptosis of alveolar epithelial cells and the lung injury caused by H1N1 infection. The findings provide a novel mechanism to protect against the H1N1-induced pulmonary injury, which may better facilitate the progress of new strategies to fight H1N1 infection or other IAV viral infections.

UiO-66 nanoparticles combat influenza A virus in mice by activating the RIG-I-like receptor signaling pathway.

The Influenza A virus (IAV) is a zoonotic pathogen that infects humans and various animal species. Infection with IAV can cause fever, anorexia, and dyspnea and is often accompanied by pneumonia characterized by an excessive release of cytokines (i.e., cytokine storm). Nanodrug delivery systems and nanoparticles are a novel approach to address IAV infections. Herein, UiO-66 nanoparticles (NPs) are synthesized using a high-temperature melting reaction. The in vitro and in vivo optimal concentrations of UiO-66 NPs for antiviral activity are 200 µg mL-1 and 60 mg kg-1, respectively. Transcriptome analysis revealed that UiO-66 NPs can activate the RIG-I-like receptor signaling pathway, thereby enhancing the downstream type I interferon antiviral effect. These NPs suppress inflammation-related pathways, including the FOXO, HIF, and AMPK signaling pathways. The inhibitory effect of UiO-66 NPs on the adsorption and entry of IAV into A549 cells is significant. This study presents novel findings that demonstrate the effective inhibition of IAV adsorption and entry into cells via UiO-66 NPs and highlights their ability to activate the cellular RIG-I-like receptor signaling pathway, thereby exerting an anti-IAV effect in vitro or in mice. These results provide valuable insights into the mechanism of action of UiO-66 NPs against IAV and substantial data for advancing innovative antiviral nanomedicine.

Remediation of cadmium contaminated soil using electrokinetic-phytoremediation system with rotary switching electrodes.

Considering both electrokinetic remediation and phytoremediation have limitations, an electrokinetic phytoremediation (EP) system was constructed to obtain efficient and environmentally friendly remediation results. This study indicates that the electric field can promote the absorption of Cd by ryegrass with little impact on soil physicochemical properties under the condition of rotary switching electrodes, and the accumulation of Cd in the aboveground and underground parts of ryegrass increased by 145.2% and 93.7%, respectively. The DC electric field combined with ryegrass under rotary switching electrode mode proved to be the optimal condition for the remediation of Cd contaminated soil with a remediation efficiency of 66.7%. Moreover, the rotary switching of the electrodes alleviated the suppression of the growth of ryegrass by the DC electric field. During the EP remediation process, the electric field promoted the transformation of the residue state of Cd to the other forms, which accelerated the desorption rate of Cd from the soil and facilitated the migration of Cd into plants. In conclusion, EP is a green and efficient remediation technology for heavy metal contaminated soil with good application prospects.

Gut commensal bacteria exacerbate toxoplasmosis associated with TgSheepCHn5 (ToxoDB#2) and TgRedpandaCHn1 (ToxoDB#20) through Th1 immune response.

Oral infection of mice with several strains of Toxoplasma gondii results in intestinal pathological lesions, which contributes to the invasion of this parasite. However, the exact mechanism is unclear, and only a few strains have been explored. Here, T. gondii TgSheepCHn5 and TgRedpandaCHn1 strains from sheep and red panda were evaluated. The TgSheepCHn5 and TgRedpandaCHn1 strains induced intestinal lesions, loss of Paneth cells, and gut commensal bacteria dysbiosis in Swiss Webster mice. The lesions and loss of Paneth cells were dependent on IFN-γ and gut commensal bacteria during T. gondii infection. Deleting IFN-γ or gut commensal bacteria suppressed the Th1 immune response, alleviated the lesions and parasite loading, and upregulated the number of Paneth cells. Loss of IFN-γ production accelerated mice death, whereas the deletion of gut commensal bacteria enhanced the survival time of the host. The Th1 cell immune responses have positive and negative effects on toxoplasmosis, resistance to T. gondii infection, and acceleration intestine lesions. Adjustment of Th1 cell responses and gut commensal bacteria may be effective treatments for toxoplasmosis.

Sarcocystis species in wild and domestic sheep (Ovis ammon and Ovis aries) from China.

BACKGROUND: Sarcocystis species are intracellular protozoan parasites that can pose a threat to animal health and food safety. The aim of this study was to investigate the prevalence of infection with Sarcocystis infection in sheep from China. RESULTS: In total, 52.51% (335/638) of tissue samples from domestic sheep contained sarcocysts through examination by light microscopy. The organisms were identified as S. tenella and S. arieticanis by molecular assays. Macroscopic S. gigantea and S. medusiformis were not found. The average sarcocysts loading was 18.07 ± 29.87 per square centimeter in the myocardium of domestic sheep. Furthermore, two specimens of argali (Ovis ammon) were examined and sarcocysts were found in the myocardium of one animal. According to the sequence of the cox1 gene of sarcocysts from argali, it was speculated as S. tenella. CONCLUSIONS: We found a high prevalence and parasite load of Sarcocystis in sheep from both central and northwest China. This report is the first to indicate that argali may be a natural intermediate host for S. tenella.

Synergistic Photothermal Tumor Immunotherapy by 1-MT Based on Zeolitic Imidazolate Framework-8 with pH-High Sensitivity.

Background: A successful immune response against tumors depends on various cellular processes. Hence, there is an urgent need to construct a proficient nanoplatform for immunotherapy that can concurrently regulate the activities of various cells participating in the immune process. We have developed zeolitic imidazolate framework-8 (ZIF-8) formula, with good pH sensitivity, which is conducive to the release of drugs in the tumor site (acidic environment) and significantly improves immunotherapy. This is achieved through the coordinated action of different therapeutic agents, such as the photothermal agent polydopamine (PDA), the chemodrug camptothecin (CPT), and the immunomodulator 1-methyl-D-tryptophan (1-MT). Materials and Methods: In this study, we evaluated the antitumor effect of PDA/(CPT + 1-MT) @ZIF-8 (PCMZ) nanoparticles (NPs) in vitro and in vivo and investigated the molecular mechanism of PCMZ NPs in tumor suppression via photothermal-chemo-immunotherapy. Results: MTT and Annexin V-FITC/PI double staining apoptosis test showed that PCMZ NPs could induce apoptosis of 4T1 cell, and PCMZ NPs could cause 4T1 cell necrosis under 808 nm laser irradiation. The objective is to establish a unilateral breast cancer model in mice and investigate the effect of PCMZ NPs on tumor growth and tumor suppression in tumor bearing mice. The results showed that PCMZ NPs showed good heating effect in vivo and effectively inhibited tumor growth under 808 nm laser irradiation. In addition, PCMZ NPs could induce the immunogenic death of tumor cells, promote the maturation of DCs, inhibit IDO pathway, and finally differentiate T cells into cytotoxic T cells and helper T cells, so as to effectively activate the anti-tumor immune response. Conclusion: The PCMZ NPs, possessing good photothermal conversion capabilities due to join of PDA, effectively overcome two main challenges in immunotherapy: insufficient stimulation of the immune response and evasion of the immune system. This provides a robust platform against invasive cancer and recurrent tumors.

PI3K/AKT-mediated autophagy inhibition facilitates mast cell activation to enhance severe inflammatory lung injury in influenza A virus- and secondary Staphylococcus aureus-infected mice.

Influenza A virus infection causes considerable morbidity and mortality each year globally, and secondary bacterial infection further exacerbates the severity and fatality of the initial viral infection. Mast cells have substantial roles in protecting the respiratory tract mucosa, while their role in viral and bacterial co-infection remains unclear. The present study revealed that secondary Staphylococcus aureus infection significantly aggravated the activation of mast cells during the initial H1N1 infection both in vivo and in vitro, which was closely related to the increased inflammatory lung injury and mortality. Meanwhile, the secondary S. aureus infection suppressed autophagy and promoted inflammatory mediators released by mast cells through activating the PI3K/Akt signaling pathway. Blocking PI3K/Akt pathway by LY294002, an inhibitor of Akt phosphorylation, could rescue autophagy and inhibit the release of inflammatory mediators. Furthermore, based on the influenza A viral and secondary bacterial infected mice model, we showed that the combination of LY294002 and antiviral drug oseltamivir could effectively reduce the inflammatory damage and pro-inflammatory cytokines releasing in lungs, recovering body weight loss and improving the survival rate from the co-infections. In conclusion, secondary bacterial infection can inhibit autophagy and stimulate mast cell activation through the PI3K/Akt pathway, which might explain why secondary bacterial infection would cause severe and fatal consequences following an initial influenza A viral infection.

Low Prevalence of Antibodies Against Toxoplasma gondii in Chinese Populations.

Toxoplasma gondii has been found to infect almost all warm-blooded animals, including humans. In this study, a total of 3,275 human serum samples were collected from hospitals in five provinces of China. About 5.13% (168/3,275) (95% CI, 4.42-5.94) of the serum samples tested positive for T. gondii IgG antibody by a modified agglutination test (MAT) (cut-off: 1:20). Significant associations were detected between geographic location (OR = 1.763), age (OR = 3.072), infertility in women (OR = 2.4409) and T. gondii infection in humans (p < 0.05). To minimize infection, citizens need to be informed about the best practices for toxoplasmosis prevention, including eating well-cooked meat, drinking boiled water, washing vegetables and fruits, and being careful during contact with cats.

Toxoplasma gondii in lambs of China: Heart juice serology, isolation and genotyping.

Toxoplasmosis is one of the most common foodborne diseases in the world. The objective of this study was to determine Toxoplasma gondii infection in lambs from Henan province, China. A total of 166 lamb hearts were collected from 2017 to 2019. T. gondii infection was determined by the Modified Agglutination Test (MAT) using heart juice of lambs. 11 isolates (TgSheepCHn3 - TgSheepCHn13) were obtained from samples with MAT titers ≥1:100. The rate of T. gondii isolation increased with antibody titer against T. gondii (P < 0.05). No isolate was obtained from samples with titer 1:25 and 1:50, suggesting the cut-off titer for MAT is better set at 1:100. With cut-off value of 1:100, IgG antibodies to T. gondii were found in 25.3% (42/166) of the lambs by MAT. T. gondii parasite was not found in IHC and HE-stained tissue sections of lamb hearts (0/166). Sixty-seven heart tissues with ≥1:25 MAT titers were subjected to acid pepsin digestion and detected T. gondii by PCR. Only 7.5% (5/67) of DNA amplified products were found in heart tissues by the primer TOX5/TOX8. Brain tissue cysts were observed in all mice infected with the 11 isolates at day 60 post infection, suggesting these isolates are non-lethal to mice. PCR-RFLP analysis revealed that 7 isolates belonged to ToxoDB#2, 4 isolates belonged to ToxoDB#4. This is the first isolation of ToxoDB#2 and ToxoDB#4 from lambs in China. Interestingly, none of these isolates belongs to the ToxoDB#9 that is common in China. Our results suggest that the genetic diversity and population structure of T. gondii from China maybe more abundant and magical than previous speculation.

Low prevalence of viable Toxoplasma gondii in swine from slaughter houses in the central of China.

BACKGROUND: Toxoplasma gondii is widely distributed and can infect many species of warm-blooded animals, including swine. This study aimed to evaluate the prevalence of T. gondii in swines from the central of China. A total of 2798 samples, including 305 hearts, 2086 diaphragms, and 407 sera were collected from different swine in Henan Province, China. The modified agglutination test was used to detect antibodies against T. gondii in sera from jugular vein blood and heart blood (cut-off: 1:25), diaphragm juice (cut-off: 1:10). T. gondii DNA was screened from the digestive fluids of all diaphragm tissue samples and seropositive hearts, and attempt to isolate viable T. gondii strain by bioassay in mice. RESULTS: A total of 9.94% (278/2798) swine tested positive for T. gondii antibodies. Region, but not gender, was associated with T. gondi seropositivity in swine. T. gondii nucleic acid was not found in the tissue digestive fluids (2090 swines). Three groups of mice showed T. gondii antibodies after having been bioassayed with diaphragm samples (n = 81, which came from 2090 swine). No viable T. gondii strain was isolated from muscle of swine. CONCLUSIONS: This is the first large-scale survey T. gondi infection in swine from the central of China. Overall, the prevalence of viable T. gondii in swine was low. Nevertheless, T. gondii infection is present in swine from the central of China. Consumers may acquire T. gondii infection from ingestion of raw or undercooked pork.

Evidence of red panda as an intermediate host of Toxoplasma gondii and Sarcocystis species.

Toxoplasma gondii has been found to infect almost all warm-blooded animals; however, some hosts lack direct evidence of T. gondii infection. The red panda (Ailurus fulgens) is an endangered species that mainly lives in temperate forests of South Asia. Here, T. gondii infection in red pandas from zoos in China were reported. Antibodies to T. gondii were found in 14.3% (2/14) of red pandas via the modified agglutination test (MAT) with a cut-off titer of 1:25. One viable T. gondii strain was isolated from tissues of red panda and designated as TgRedpandaCHn1. DNA from tachyzoites obtained from cell culture was characterized by PCR-RFLP with 10 markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) and virulence genes of ROP5 and ROP18. The results indicate that this isolate belonged to ToxoDB genotype #20. The ROP18/ROP5 genotype combination predicated that this strain is non-lethal to mice, which is supported by the infection in mice. T. gondii tissue cysts were readily formed and mice survived. Tissue cysts observed in the histopathological sections of the tongue and diaphragm of one red panda were speculated as sarcocysts, but not T. gondii base on morphological characteristics. To our knowledge, this study is the first to report on the isolation of T. gondii from red panda. Additionally, this report provides direct evidence of red panda as an intermediate host of T. gondii and Sarcocystis species.

Isolation, genotyping and pathogenicity of a Toxoplasma gondii strain isolated from a Serval (Leptailurus serval) in China.

Toxoplasma gondii typically causes lifelong chronic infection and has been identified in a variety of intermediate and definitive hosts. Felids are capable of serving as both intermediate and definite hosts for T. gondii infection. However, there is no direct evidence to prove that servals are the intermediate host of T. gondii. In this study, T. gondii antibodies were detected in a serval by a modified agglutination test (titer, 1:200). Viable T. gondii was isolated from the striated muscles of the serval. This strain was further propagated in cell culture and designated as TgServalCHn1. Genetic characterization of DNA derived from cell culture was performed by RFLP-PCR of 10 markers, as well as polymorphic ROP5 and ROP18 genes. Results showed that this strain of T. gondii belonged to the genotype ToxoDB#20. The ROP5 allele 4 and ROP18 allele 3 suggested that this strain was avirulent, which was further supported by infection in mice. Encephalitis, immune organ necrosis and focal mononuclear cell infiltration in multiple organs were the main pathology characteristics observed in BALB/C mice infected with the TgServalCHn1 strain. To our knowledge, the present study is the first to report the isolation of T. gondii from a serval, which gives direct evidence for servals serving as an intermediate host of T. gondii. The genotyping results revealed the presence of genotype ToxoDB#20 in central China, enriching the scope of the distribution of T. gondii genotypes in Asia.

Toxoplasma gondii in four captive kangaroos (Macropus spp.) in China: Isolation of a strain of a new genotype from an eastern grey kangaroo (Macropus giganteus).

Marsupials are highly susceptible to Toxoplasma gondii infection. Here, we report T. gondii infection in four kangaroos from a zoo in China. Kangaroos were imported into China in 2000 and were since bred in zoo. In 2017-2018, four kangaroos died due to respiratory system disease or injury. The bodies were submitted to the laboratory to test for T. gondii infection. Antibodies to T. gondii were found in 75% (3/4) of the kangaroos via the modified agglutination test with the cut-off 1:25. Cysts were observed in the histopathological sections of tongue and diaphragm or squashes of fresh myocardium in two kangaroos. These cysts were confirmed as T. gondii by immunohistochemical staining and molecular biological analysis. One viable T. gondii strain was isolated from one kangaroo and designated as TgRooCHn1. DNA from T. gondii tachyzoites obtained from cell culture was characterized by 10 PCR-RFLP markers and the virulence genes ROP5 and ROP18. The genotype of this isolate did not match with any known genotypes; it was designated as ToxoDB#292. The virulence of TgRooCHn1 (104 tachyzoites) was non-lethal to mice, and it formed tissue cysts. To our knowledge, the present study is the first isolation of ToxoDB#292 strain from kangaroo. Improvemets for captive settings were initiated, including greater attention being paied to birds and stray cats, fed frozen meat for carnivores.

Direct evidence of an extra-intestinal cycle of Toxoplasma gondii in tigers (Panthera tigris) by isolation of viable strains.

Toxoplasmosis is one of the most common zoonotic diseases in the world. Felines excrete environmentally resistant Toxoplasma gondii oocysts. However, there is no direct evidence to prove tigers are the intermediate host of T. gondii. Here, we show that, IgG antibodies to T. gondii in 80% (8/10) of captive tigers. Two viable T. gondii strains (ToxoDB genotype #9) were isolated by bioassay in mice using striated muscles of two tigers (Tiger#3 and Tiger#8). Additionally, mice were confirmed as T. gondii-positive by bioassay of feces #89-110, but no viable T. gondii strain was isolated successfully. The fecal samples from tigers may contain T. gondii oocysts. This is the first report of T. gondii isolation from tigers. These results provide direct evidence that an extra-intestinal cycle of T. gondii may develop in tigers.

High prevalence of Sarcocystis spp. infections in cattle (Bos taurus) from central China.

Myocardium and diaphragm samples of cattle (n = 521) from HeNan Province (China) were screened for Sarcocystis sarcocysts by histological examination, pepsin digestion, and molecular assays. Morphology and molecular assays were used for identification. The prevalence of Sarcocystis infection in cattle was 41.5% (216/521). Histological examination identified sarcocysts in the myocardium (49.4%, 200/405) and diaphragm (13.8%, 16/116) of cattle. Two species were identified, namely S. cruzi (41.3%, 215/521) and S. hominis (0.2%, 1/521). The findings of the present study indicate a high prevalence of S. cruzi infection in cattle from central China.

Prevalence, Risk Factors, and Genotypes of Toxoplasma gondii in Food Animals and Humans (2000-2017) From China.

Toxoplasma gondii as a food-borne pathogen, the infection of it in food animals has relation with human toxoplasmosis, but the trends and epidemiological features of T. gondii infections in food animals are rarely studied in China. The aimed of this study was to assess the epidemiology and risks of T. gondii in sheep, goats, swines, chickens, yaks, cattle and humans from 2000 to 2017 and to explore prevention and control strategies. The overall seroprevalence of T. gondii infections in food animals is 23.7% (39,194/165,417, 95%CI, 23.49-23.90%), which is significantly higher than that in humans (8.2%, 95%CI, 8.06-8.39%, 8,502/103,383) (P < 0.0001). Compared the prevalence of T. gondii infections in animals and humans sampled from 2000 to 2010, it was significantly increased in the period 2011 to 2017 (P < 0.0001). Compared the food animals from non-Yangtze River, animals from regions of the Yangtze River have high seroprevalence rates for T. gondii (P < 0.0001). Furthermore, samples from the western to eastern regions of the Yellow River showed an increase in seroprevalence for T. gondii (P < 0.0001). It was speculated that T. gondii oocysts may be transmitted by water and annual precipitation possible help the oocyst spread and retain accessible for potential hosts. Effective prevention and control strategies are including water filtration or water boiling, inactivating oocysts from feline's feces, monitoring birds and rodents. Chinese 1 (ToxoDB#9) is the predominant genotype in food animals from China.

Studies on the properties of graphene oxide-alkaline protease bio-composites.

Graphene oxide (GO) nanosheets as functional material have a unique planar structure and intriguing mechanical that have attracted intensive interests recently. A method was developed for the immobilization protease on GO sheets using glutaraldehyde as cross-linking reagent. The results showed that the thermostability and reusability of immobilized protease have been obviously improved compared to the free enzyme. However, there was no significant change in optimum pH value between the free and immobilized protease. The immobilized protease exhibited good operational stability. The apparent K(m) and V(max) for free and immobilized alkaline protease were determined, and the bio-catalytic activity was not impaired by immobilization.

Supported cobalt oxide on graphene oxide: highly efficient catalysts for the removal of Orange II from water.

The current paper investigated the removal of the azo dye Orange II from water using advanced oxidation processes based on sulfate radicals. The cobalt oxide catalyst immobilized on graphene oxide (GO) can activate peroxymonosulfate (PMS) for the degradation of Orange II in water. The Co(3)O(4)/GO catalyst system was characterized via X-ray diffraction, Fourier transform infrared spectroscopy, Raman spectroscopy, scanning electron microscopy, and X-ray spectroscopy. Results showed that Co(3)O(4) was distributed on GO. The Co(3)O(4)/GO catalyst system exhibited high activity in Orange II oxidation when the Co(3)O(4)/GO catalyst has an optimum Co(3)O(4) loading. In addition, 100% decomposition could be achieved within 6 min with 0.2mM Orange II, 0.1 g L(-1) catalyst, and 2mM PMS. Meanwhile, inductively coupled plasma analysis revealed that the leach of cobalt ions was low. The catalyst also exhibited stable performance after several rounds of regeneration. Several operational parameters, such as catalyst amount, oxidant amount, pH, temperature, and oxidation rate, affected the degradation of Orange II.