Differential immune responses to Segniliparus rotundus and Segniliparus rugosus infection and analysis of their comparative virulence profiles.

Two closely related bacterial species, Segniliparus rotundus and Segniliparus rugosus, have emerged as important human pathogens, but little is known about the immune responses they elicit or their comparative pathophysiologies. To determine the virulence and immune responses of the two species, we compared their abilities to grow in phagocytic and non-phagocytic cells. Both species maintained non-replicating states within A549 epithelial cells. S. rugosus persisted longer and multiplied more rapidly inside murine bone marrow-derived macrophages (BMDMs), induced more pro-inflammatory cytokines, and induced higher levels of macrophage necrosis. Activation of BMDMs by both species was mediated by toll-like receptor 2 (TLR2), followed by mitogen-activated protein kinases (MAPK) and nuclear factor κB (NF-κB) signaling pathways, indicating a critical role for TLR2 in Segniliparus-induced macrophage activation. S. rugosus triggered faster and stronger activation of MAPK signaling and IκB degradation, indicating that S. rugosus induces more pro-inflammatory cytokines than S. rotundus. Multifocal granulomatous inflammations in the liver and lung were observed in mice infected with S. rugosus, but S. rotundus was rapidly cleared from all organs tested within 15 days post-infection. Furthermore, S. rugosus induced faster infiltration of innate immune cells such as neutrophils and macrophages to the lung than S. rotundus. Our results suggest that S. rugosus is more virulent and induces a stronger immune response than S. rotundus

Consoramides A–C, New Zwitterionic Alkaloids from the Fungus Irpex consors

In our ongoing search for new secondary metabolites from fungi, a basidiomycete fungus Irpex consors was selected for mycochemical investigation, and three new zwitterionic alkaloids (1-3) and five known compounds (4-8) were isolated from the culture broth (16 l) of I. consors. The culture filtrate was fractionated by a series of column chromatography including Diaion HP-20, silica gel, and Sephadex LH-20, Sep-Pak C18 cartridge, medium pressure liquid chromatography (MPLC), and high pressure liquid chromatography (HPLC) to yield eight compounds (1-8). The structures of the isolated compounds were elucidated by the interpretation of nuclear magnetic resonance (NMR) spectra and high-resolution mass spectrometry (HR-MS). Their antioxidant and antibacterial activities were examined. The zwitterionic structures of three new sesquiterpene alkaloids (1-3) were determined together with five known compounds identified as stereumamide E (4), stereumamide G (5), stereumamide H (6), stereumamide D (7), and sterostrein H (8). This is the first report of the zwitterionic alkaloids in the culture broth of I. consors. Three new zwitterionic alkaloids were named as consoramides A–C (1-3)

Clinical Usability of Embryo Development Using a Combined Qualitative and Quantitative Approach in a Single Vitrified-Warmed Blastocyst Transfer: Assessment of Pre-Vitrified Blastocyst Diameter and Post-Warmed Blastocyst Re-Expansion Speed

Improving the safety and efficacy of assisted reproductive technology programs has been a continuous challenge. Traditionally, morphological grading has been used for embryo selection. However, only a few studies have assessed the morphokinetic variables and morphological dynamics of blastocysts. In the present study, we aimed to perform a quantitative analysis of blastocyst diameter and re-expansion speed. This in-depth morphokinetic evaluation can correlate with currently observed pregnancy outcomes. In total, 658 single vitrified-warmed blastocyst transfer cycles were performed between October 2017 and December 2021, which were divided into four groups according to the pre-vitrified blastocyst diameter. After warming, the groups were subdivided according to the blastocyst re-expansion speed. These quantitative measurements were performed using a time-lapse system. Both diameter and speed are essential in determining the blastocyst quality, while age, day of freezing, and blastocyst quality are crucial from a clinical perspective. The application of both quantitative (diameter and speed) and qualitative (blastocyst quality scores) parameters can help evaluate the clinical usability of blastocysts. This method can prove useful for embryologists in counseling their patients and determining pregnancy patient-oriented strategies

Levels of humoral responses in C57BL/6 mice following infection with <i>S. rugosus</i> and <i>S. rotundus</i> at designated times.

<p>Antibody responses of IgG1 (A), IgG_2a (B) and the IgG_2a/IgG1 ratio (C), IL-4 (D), and IL-17 (E) were measured by ELISA as described in “Materials and Methods”. Each sample was examined in triplcate. Data are presented as the absorbance at 450 nm and are representative of three experiments. The results are presented as means ± SD of each group (<i>n</i> = 6 per group, *<i>p</i><0.05, **<i>p</i><0.01).</p

Quantitative analysis of necrosis and apoptosis in BMDMs infected with <i>S. rugosus</i> or <i>S. rotundus.</i>

<p>A) Apoptotic and necrotic cell death measured by 7-ADD and Annexin V assay. BMDMs were infected with <i>S. rugosus</i> or <i>S. rotundus</i> at a MOI of 1 or 10 for 24 h. After incubation, cells were collected and Annexin V-PE and 7-AAD were added. Samples were analyzed using flow cytometry. Results represent mean percentage of positive cells ± SD of all experiments performed. This experiment was repeated three times with similar results. B) LDH released from BMDMs infected with <i>S. rugosus</i> and <i>S. rotundus</i> at MOI of 1 according to the days post-infection. C) LDH released from BMDMs infected with <i>S. rugosus</i> and <i>S. rotundus</i> at MOI of 1 or 10 for 24 h. Culture supernatants were harvested and the release of LDH was assessed using a cytotoxicity detection kit. This experiment was repeated three times with similar results (*<i>p</i><0.05 or **<i>p</i><0.01).</p