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Objective To study the effect of spermatogenic cell-specific promoter Dazl in Spermatogenic cells. Constructing the eukaryotic reporter gene expression vector AAV-Dazl-RFP-Flag containing spermatogenic cell-specific promoter Dazl. Methods The Adeno-associated virus vector AAV-CMV-RFP-Flag was modified by homologous recombination to construct mouse permatogenic cell-specific expression vector AAV-Dazl-RFP-Flag. In vitro, GC1-spg wild-type cells and HEK-293T cells were transfected, and the expression specificity of AAV-Dazl-RFP-Flag in vitro was verified by immunofluorescence, q-PCR, and Western blot. In vivo, AAV-Dazl-RFP-Flag was packaged with adeno-associated virus (AAV), and AAV was injected into the testes of mouse through microinjection technology, then the expression specificity of AAV-Dazl-RFP-Flag in vivo was detected by q-PCR. Results Mouse Sper- matogenic cell-specific expression vector AAV-Dazl-RFP-Flag is specifically expressed in spermatogenic, cells thus provid providing a more efficient tool for gene therapy of male reproductive infertility.

Abstract Background Acinetobacter baumannii, a significant nosocomial pathogen, has evolved resistance to almost all conventional antimicrobial drugs. Bacteriophage therapy is a potential alternative treatment for multidrug-resistant bacterial infections. In this study, one lytic bacteriophage, ZZ1, which infects A. baumannii and has a broad host range, was selected for characterization. Results Phage ZZ1 and 3 of its natural hosts, A. baumanni clinical isolates AB09V, AB0902, and AB0901, are described in this study. The 3 strains have different sensitivities to ZZ1, but they have the same sensitivity to antibiotics. They are resistant to almost all of the antibiotics tested, except for polymyxin. Several aspects of the life cycle of ZZ1 were investigated using the sensitive strain AB09V under optimal growth conditions. ZZ1 is highly infectious with a short latent period (9 min) and a large burst size (200 PFU/cell). It exhibited the most powerful antibacterial activity at temperatures ranging from 35°C to 39°C. Moreover, when ZZ1 alone was incubated at different pHs and different temperatures, the phage was stable over a wide pH range (4 to 9) and at extreme temperatures (between 50°C and 60°C). ZZ1 possesses a 100-nm icosahedral head containing double-stranded DNA with a total length of 166,682 bp and a 120-nm long contractile tail. Morphologically, it could be classified as a member of the Myoviridae family and the Caudovirales order. Bioinformatic analysis of the phage whole genome sequence further suggested that ZZ1 was more likely to be a new member of the Myoviridae phages. Most of the predicted ORFs of the phage were similar to the predicted ORFs from other Acinetobacter phages. Conclusion The phage ZZ1 has a relatively broad lytic spectrum, high pH stability, strong heat resistance, and efficient antibacterial potential at body temperature. These characteristics greatly increase the utility of this phage as an antibacterial agent; thus, it should be further investigated.

Systemic inflammatory response syndrome plays an important role in the development of sepsis. GABAergic and cholinergic pathways activation are considered important for inflammatory response regulation. Tumor necrosis factor (TNF)-[alpha], interleukin (IL)-1[beta], IL-12, IL-10, as well as inducible nitric oxide synthase (iNOS)-derived nitric oxide (NO) are important inflammatory mediators involved in the pathogenesis of sepsis. Muscimol, an active compound from the mushroom Amanita muscaria (L.) Lam., is a potent [GABA.sub.A] agonist, inhibits inflammatory response via activating [GABA.sub.A] receptor and vagus nerve. However, the effect of muscimol on lipopolysaccharide (LPS)-induced systemic inflammatory response is still unclear. Therefore, we studied the effects of muscimol on systemic inflammatory response and survival rate in endotoxemic mice. Mice endotoxemia was induced by LPS. Muscimol was given to mice or RAW264.7 cells 30 min before LPS (10 mg/kg, i.p., or 10 ng/mL, respectively). Mice received GABAergic and cholinergic receptor antagonists 30 min before muscimol and LPS. Muscimol decreased TNF- [alpha], IL-1[beta], IL-12, iNOS-derived NO, and increased IL-10 levels and survival rate after LPS treatment. Muscimol significantly decreased nuclear factor kappa B (NF-[kappa]B) activity, increased I[kappa]B expression, and decreased pIKK expression in LPS- treated RAW264.7 cells. GABAergic and cholinergic antagonists failed to reverse muscimol's protection in LPS-treated mice. In conclusion, muscimol protected against systemic inflammatory response in endotoxemic mice may be partially independent of GABAergic and cholinergic receptors. Key words: muscimol, systemic inflammatory response, LPS, GABA receptor, cholinergic receptor Le syndrome de reaction inflammatoire generalisee joue un role important dans le developpement de la septicemie. L'activation des voies de signalisation GABAergiques et cholinergiques est consideree comme etant importante dans la regulation de la reaction inflammatoire. Le facteur de necrose tumorale (TNF)-, l'interleukine (IL)-1[beta], l'IL-12, l'IL-10, de meme que l'oxyde nitrique (NO) derive de la NO synthase inductible (iNOS) sont d'importants mediateurs inflammatoires, qui jouent un role dans la pathogenese de la septicemie. Le muscimol, un compose actif issu du champignon Amanita muscaria, puissant agoniste des recepteurs GABAA, permet d'inhiber la reaction inflammatoire par l'intermediaire de l'activation des recepteurs GABAA et du nerf vague. Cependant, l'effet du muscimol sur la reaction inflammatoire generalisee engendree par les lipopolysaccharides (LPS) demeure a clarifier. Par consequent, nous avons etudie les effets du muscimol sur la reaction inflammatoire generalisee et sur le taux de survie chez la souris endotoxemique. Nous avons obtenu une endotoxemie chez des souris a l'aide de LPS. Nous avons administre du muscimol chez des souris ou chez des cellules RAW264.7 30 minutes avant les LPS (10 mg/kg, i.p., ou 10 ng/ml, respectivement). Les souris ont recu des antagonistes GABAergiques et cholinergiques 30 minutes avant le muscimol et les LPS. Apres l'administration des LPS, le muscimol a entraine une diminution des taux de TNF-[alpha], d'IL-1[beta], d'IL-12, de NO derive de l'iNOS, avec une diminution des taux d'IL-10 de meme que du taux de survie. Le muscimol entrainait une diminution marquee de l'activite du NF-[kappa]B, une augmentation de l'activite de I'I[kappa]B, ainsi qu'une diminution de l'expression de pIKK dans les cellules RAW264.7 exposees aux LPS. Les antagonistes GABAergiques et cholinergiques n'ont pas permis de contrer la protection qu'engendrait le muscimol chez les souris auxquelles des LPS etaient administres. En conclusion, la protection contre la reaction inflammatoire generalisee obtenue avec le muscimol chez des souris endotoxemiques pourrait etre partiellement independante des recepteurs GABAergiques et cholinergiques. [Traduit par la Redaction] Mots-cles : muscimol, reaction inflammatoire generalisee, LPS, recepteurs du GABA, recepteur cholinergique, Introduction Systemic inflammatory response syndrome (SIRS), mainly caused by Gram-negative bacteria, plays a vital role in the development of sepsis (Kaukonen et al. 2015; Chakraborty and Burns 2021). It estimates [...]

[Objectives] To clarify the influence of sex, age (days) and temperature, on the flight ability of adult Hyphantria cunea, and there by provide information essential for monitoring, and predicting, the dispersal of this pest. [Methods] The effect of gender, age and temperature on the flight ability of adult H. cunea was measured in an insect flight mill. [Results] Males were significantly stronger fliers than females with a maximum flight distance of 17.87 km and cumulative flight time of 17.38 h. One day-old males had the longest cumulative flight distance; the average cumulative flight distance and average cumulative flight time of males decreased with age. The maximum female flight distance was 8.21 km and the longest cumulative flight time was 7.4 h. Female flight distance tended to first increase, then decrease, with age. Three day-old females had the maximum cumulative flight distance. A two-way ANOVA of the average cumulative flight distance, average cumulative flight time, and average flight speed, did not detect a significant interaction between sex and age. Male flight speed was significantly slower at a temperature of 16 °C. [Conclusion] Male H. cunea are stronger fliers than females. Male flight ability gradually decreases with age, whereas that of females tends to increase up to 3 days of age, then decrease. Low temperatures reduce the flight speed of H. cunea males. [ABSTRACT FROM AUTHOR]