Protective effect of arctiin against Toxoplasma gondii HSP70-induced allergic acute liver injury by disrupting the TLR4-mediated activation of cytosolic phospholipase A2 and platelet-activating factor.

Toxoplasma gondii (T. gondii)-derived heat shock protein 70 (T.g.HSP70) is a toxic protein that downregulates host defense responses against T. gondii infection. T.g.HSP70 was proven to induce fatal anaphylaxis in T. gondii infected mice through cytosolic phospholipase A2 (cPLA2) activated-platelet-activating factor (PAF) production via Toll-like receptor 4 (TLR4)-mediated signaling. In this study, we investigated the effect of arctiin (ARC; a major lignan compound of Fructus arctii) on allergic liver injury using T.g.HSP70-stimulated murine liver cell line (NCTC 1469) and a mouse model of T. gondii infection. Localized surface plasmon resonance, ELISA, western blotting, co-immunoprecipitation, and immunofluorescence were used to investigate the underlying mechanisms of action of ARC on T. gondii-induced allergic acute liver injury. The results showed that ARC suppressed the T.g.HSP70-induced allergic liver injury in a dose-dependent manner. ARC could directly bind to T.g.HSP70 or TLR4, interfering with the interaction between these two factors, and inhibiting activation of the TLR4/mitogen-activated protein kinase/nuclear factor-kappa B signaling, thereby inhibiting the overproduction of cPLA2, PAF, and interferon-γ. This result suggested that ARC ameliorates T.g.HSP70-induced allergic acute liver injury by disrupting the TLR4-mediated activation of inflammatory mediators, providing a theoretical basis for ARC therapy to improve T.g.HSP70-induced allergic liver injury.

ATL Protein Family: Novel Regulators in Plant Response to Environmental Stresses.

Plants actively develop intricate regulatory mechanisms to counteract the harmful effects of environmental stresses. The ubiquitin-proteasome pathway, a crucial mechanism, employs E3 ligases (E3s) to facilitate the conjugation of ubiquitin to specific target substrates, effectively marking them for proteolytic degradation. E3s play critical roles in many biological processes, including phytohormonal signaling and adaptation to environmental stresses. Arabidopsis Toxicosa en Levadura (ATL) proteins, belonging to a subfamily of RING-H2 E3s, actively modulate diverse physiological processes and plant responses to environmental stresses. Despite studies on the functions of certain ATL family members in rice and Arabidopsis, most ATLs still need more comprehensive study. This review presents an overview of the ubiquitin-proteasome system (UPS), specifically focusing on the pivotal role of E3s and associated enzymes in plant development and environmental adaptation. Our study seeks to unveil the active modulation of plant responses to environmental stresses by E3s and ATLs, emphasizing the significance of ATLs within this intricate process. By emphasizing the importance of studying the roles of E3s and ATLs, our review contributes to developing more resilient plant varieties and promoting sustainable agricultural practices while establishing a research roadmap for the future.

Poly(1,3-Propylene Glycol Citrate) as a Plasticizer for Toughness Enhancement of Poly-L-Lactic Acid.

Despite the unique features of poly-L-lactic acid (PLLA), its mechanical properties, such as the elongation at break, need improvement to broaden its application scope. Herein, poly(1,3-propylene glycol citrate) (PO3GCA) was synthesized via a one-step reaction and evaluated as a plasticizer for PLLA films. Thin-film characterization of PLLA/PO3GCA films prepared via solution casting revealed that PO3GCA shows good compatibility with PLLA. The addition of PO3GCA slightly improves the thermal stability and enhances the toughness of PLLA films. In particular, the elongation at break of the PLLA/PO3GCA films with PO3GCA mass contents of 5%, 10%, 15%, and 20% increases to 172%, 209%, 230%, and 218%, respectively. Therefore, PO3GCA is promising as a plasticizer for PLLA.

Coixol ameliorates Toxoplasma gondii infection-induced lung injury by interfering with T. gondii HSP70/TLR4/NF-κB signaling pathway.

Toxoplasma gondii (T. gondii) is an obligate intracellular protozoan parasite that causes pulmonary toxoplasmosis, although its pathogenesis is incompletely understood. There is no cure for toxoplasmosis. Coixol, a plant polyphenol extracted from coix seeds, has a variety of biological activities. However, the effects of coixol on T. gondii infection have not been clarified. In this study, we infected a mouse macrophage cell line (RAW 264.7) and BALB/c mice with the T. gondii RH strain to establish infection models in vitro and in vivo, respectively, to explore protective effects and potential mechanisms of coixol on lung injury caused by T. gondii infection. Anti-T. gondii effects and underlying anti-inflammatory mechanisms of coixol were investigated by real-time quantitative PCR, molecular docking, localized surface plasmon resonance, co-immunoprecipitation, enzyme-linked immunosorbent assay, western blotting, and immunofluorescence microscopy. The results show that coixol inhibits T. gondii loads and T. gondii-derived heat shock protein 70 (T.g.HSP70) expression. Moreover, coixol reduced inflammatory cell recruitment and infiltration, and ameliorated pathological lung injury induced by T. gondii infection. Coixol can directly bind T.g.HSP70 or Toll-like receptor 4 (TLR4) to disrupt their interaction. Coixol prevented overexpression of inducible nitric oxide synthase, tumor necrosis factor-α, and high mobility group box 1 by inhibiting activation of the TLR4/nuclear factor (NF)-κB signaling pathway, consistent with effects of the TLR4 inhibitor CLI-095. These results indicate that coixol improves T. gondii infection-induced lung injury by interfering with T.g.HSP70-mediated TLR4/NF-κB signaling. Altogether, these findings suggest that coixol is a promising effective lead compound for the treatment of toxoplasmosis.

OsCSN1 regulates the growth of rice seedlings through the GA signaling pathway in blue light.

Blue light can regulate the photomorphogenesis of plants through blue light receptors to influence seedling growth and development. The COP9 signaling complex (CSN), a vital regulator of photomorphogenesis, is a highly conserved protein complex. CSN1 is the largest and most critical subunit in the CSN with a complex N-terminal function that supports most of the functions of CSN1 and is mainly involved in plant growth and development processes. The CSN is also required in the blue light-mediated photomorphogenesis response of seedlings. In this study, the OsCSN1 subunit of Oryza sativa subsp. japonica (rice) was edited and screened, and OsCSN1 deletion mutant, OsCSN1 weak expression mutant and OsCSN1 overexpression mutant were constructed. The mechanism of OsCSN1 and its N-terminal effects on rice seedling growth and development under blue light conditions were investigated. The addition of exogenous hormone gibberellin (GA3) and gibberellin synthesis inhibitor paclobutrazol (PAC) caused aboveground phenotypic and protein (such as CUL4 and SLR1) changes. Blue light regulates the degradation of SLR1 through OsCSN1, which regulates the growth and development of rice seedling height, the first incomplete leaf, and the coleoptile. It is hypothesized that rice affects CRY-COP1 interactions after sensing blue light signals through the cryptochrome, and the nuclear localization of COP1 is regulated by the CSN complex. OsCSN1 is a negative regulator in response to blue light. The core structural domain of action that inhibits the growth of the aboveground part of rice seedlings is located at the N-terminal of OsCSN1. OsCSN1 regulates the nuclear localization of COP1 through the COP9 signaling complex and degrades SLR1 through CUL4-based E3 ligase. Ultimately, it affects the synthesis of the endogenous hormone GA, thereby inhibiting the aboveground growth and development of rice seedlings.

Protective effect of ginsenoside Rh2 against Toxoplasma gondii infection-induced neuronal injury through binding TgCDPK1 and NLRP3 to inhibit microglial NLRP3 inflammasome signaling pathway.

BACKGROUND: Toxoplasma gondii (T. gondii) is a neurotropic obligate intracellular parasite that can activate microglial and promote neuronal apoptosis, leading to central nervous system diseases. The NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome signaling complex plays a key role in inducing neuroinflammation. Our previous studies have found that ginsenoside Rh2 (GRh2) inhibits T. gondii infection-induced microglial activation and neuroinflammation by downregulating the Toll-like receptor 4/nuclear factor-kappa B signaling pathway. However, whether GRh2 reduces T. gondii infection-induced neuronal injury through actions on microglial NLRP3 inflammasome signaling has not yet been clarified. METHODS: In this study, we employed T. gondii RH strain to establish in vitro and in vivo infection models in BV2 microglia cell line and BALB/c mice. Molecular docking, localized surface plasmon resonance assay, quantitative competitive-PCR, ELISA, western blotting, flow cytometric analysis, and immunofluorescence were performed. RESULTS: Our results showed that GRh2 alleviated neuropathological damage and neuronal apoptosis in cortical tissue of T. gondii-infected mice. GRh2 and CY-09 (an inhibitor of NLRP3) exhibited potent anti-T. gondii effects through binding T. gondii calcium-dependent protein kinase 1 (TgCDPK1). GRh2 decreased Iba-1 (a specific microglial marker) and NLRP3 inflammasome signaling pathway-related protein expression by binding NLRP3. Co-culture of microglia/primary cortical neurons revealed that T. gondii-induced microglial activation caused neuronal apoptosis, but GRh2 reduced this effect, consistent with the effects of CY-09. CONCLUSION: Taken together, our results show that GRh2 has a protective effect against T. gondii infection-induced neuronal injury by binding TgCDPK1 and NLRP3 to inhibit NLRP3 inflammasome signaling pathway in microglia.

Ginsenoside Rh2 reduces depression in offspring of mice with maternal toxoplasma infection during pregnancy by inhibiting microglial activation via the HMGB1/TLR4/NF-κB signaling pathway.

BACKGROUND: Maternal Toxoplasma gondii (T. gondii) infection during pregnancy has been associated with various mental illnesses in the offspring. Ginsenoside Rh2 (GRh2) is a major bioactive compound obtained from ginseng that has an anti-T. gondii effect and attenuates microglial activation through toll-like receptor 4 (TLR4)/nuclear factor-kappa B (NF-κB) signaling pathway. GRh2 also alleviated tumor-associated or lipopolysaccharide-induced depression. However, the effects and potential mechanisms of GRh2 on depression-like behavior in mouse offspring caused by maternal T. gondii infection during pregnancy have not been investigated. METHODS: We examined GRh2 effects on the depression-like behavior in mouse offspring, caused by maternal T. gondii infection during pregnancy, by measuring depression-like behaviors and assaying parameters at the neuronal and molecular level. RESULTS: We showed that GRh2 significantly improved behavioral measures: sucrose consumption, forced swim time and tail suspended immobility time of their offspring. These corresponded with increased tissue concentrations of 5-hydroxytryptamine and dopamine, and attenuated indoleamine 2,3-dioxygenase or enhanced tyrosine hydroxylase expression in the prefrontal cortex. GRh2 ameliorated neuronal damage in the prefrontal cortex. Molecular docking results revealed that GRh2 binds strongly to both TLR4 and high mobility group box 1 (HMGB1). CONCLUSION: This study demonstrated that GRh2 ameliorated the depression-like behavior in mouse offspring of maternal T. gondii infection during pregnancy by attenuating the excessive activation of microglia and neuroinflammation through the HMGB1/TLR4/NF-κB signaling pathway. It suggests that GRh2 could be considered a potential therapy in preventing and treating psychiatric disorders in the offspring mice of mothers with prenatal exposure to T. gondii infection.

New role of sertraline against Toxoplasma gondii-induced depression-like behaviours in mice.

Toxoplasma gondii (T. gondii) is a neurotropic protozoan parasite, which can cause mental and behavioural disorders. The present study aimed to elucidate the effects and underlying molecular mechanisms of sertraline (SERT) on T. gondii-induced depression-like behaviours. In the present study, a mouse model and a microglial cell line (BV2 cells) model were established by infecting with the T. gondii RH strain. In in vivo and in vitro experiments, the underlying molecular mechanisms of SERT in inhibiting depression-like behaviours and cellular perturbations caused by T. gondii infection were investigated in the mouse brain and BV2 cells. The administration of SERT significantly ameliorated depression-like behaviours in T. gondii-infected mice. Furthermore, SERT inhibited T. gondii proliferation. Treatment with SERT significantly inhibited the activation of microglia and decreased levels of pro-inflammatory cytokines such as tumour necrosis factor-alpha, and interferon-gamma, by down-regulating tumour necrosis factor receptor 1/nuclear factor-kappa B signalling pathway, thereby ameliorating the depression-like behaviours induced by T. gondii infection. Our study provides insight into the underlying molecular mechanisms of the newly discovered role of SERT against T. gondii-induced depression-like behaviours.

Resveratrol modulates Toxoplasma gondii infection induced liver injury by intervening in the HMGB1/TLR4/NF-κB signaling pathway.

Toxoplasma gondii (T. gondii) is an obligate intracellular parasite that can cause liver diseases in the host, including hepatitis and hepatomegaly. High mobility group box 1 (HMGB1) is the main inflammatory mediator causing cell injury or necrosis. HMGB1 binds to toll like receptor 4 (TLR4), then activates the nuclear factor-κB (NF-κB) signaling pathway, which promotes the release of inflammatory factors. Our previous studies showed that HMGB1 mediated TLR4/NF-κB signaling pathway plays an important role in liver injury induced by T. gondii infection. Resveratrol (RSV) is a small polyphenol, which has anti-inflammatory, anti-cancer, anti-T. gondii effect. However, the effect of RSV on liver injury caused by T. gondii infection is unclear. This study used the RH strain tachyzoites of T. gondii to infect murine liver line, NCTC-1469 cells to establish an in vitro model and acute infection of mice for the in vivo model to explore the protective effect of RSV on liver injury induced by T. gondii infection. The results showed that RSV inhibited the proliferation of T. gondii in the liver, reduced the alanine aminotransferase/aspartate aminotransferase levels and pathological liver damage. Additionally, RSV inhibited the production of tumor necrosis factor-α, inducible nitric oxide synthase and HMGB1 by interfering with the TLR4/NF-κB signaling pathway. These results indicate that RSV can protect liver injury caused by T. gondii infection by intervening in the HMGB1/TLR4/NF-κB signaling pathway. This study will provide a theoretical basis for RSV treatment of T. gondii infection induced liver injury.

Cross-infection of adenovirus among medical staff: A warning from the intensive care unit in a tertiary care teaching hospital in China.

RATIONALE: In 2019, a small HAdV55-associated outbreak of adenovirus infection occurred among the intensive care unit (ICU) staff in Xiangya Hospital of Central South University in Hunan Province, China, during the treatment of a patient. OBJECTIVE: To investigate the characteristics of a nosocomial adenovirus outbreak in an ICU. METHODS: We evaluated all the patients treated and the medical staff working in the ICU from August 1 to September 4, 2019. We further performed an epidemiological and molecular analysis for this outbreak from patient to healthcare workers and between healthcare workers. After the outbreak, we adopted exposure prevention and droplet prevention measures based on standard precautions. MEASUREMENTS AND MAIN RESULTS: Between August 1 and August 27, 2019, 27 cases of human adenovirus cross-infection were reported in our institution. Among the cases, eleven were doctors (41%), eleven were nurses (41%), three were respiratory therapists (11%), and two were caregivers (7%). The attack rate was 28.4%, and the fatality rate was 0. The results showed that contact with the index case, lack of hand hygiene or gloving adherence were risk factors for infection after adenovirus exposure. After taking specific precautions, no new cases of infection have appeared since August 27. CONCLUSIONS: Our results show that HAdV55 in a single patient had strong transmission potential in an intensive care unit with adequate facilities and standardized operation. We provide convincing evidence indicating that attention could be highlighted on the role of standard and specific precautions for controlling the spread of adenovirus in ICUs.

Impact of miR-223-3p and miR-2909 on inflammatory factors IL-6, IL-1ß, and TNF-α, and the TLR4/TLR2/NF-κB/STAT3 signaling pathway induced by lipopolysaccharide in human adipose stem cells.

MicroRNAs (miRNAs) are small non-coding RNA molecules that play an important role in the regulation of gene expression related to inflammatory responses. Human adipose stem cells are characterized by pluripotent differentiation potential and isolated from adipose tissues. These cells regulate inflammation mainly by interacting with immune cells and affecting the secretion of immune factors; details of this interaction are currently unknown. In the current study, we successfully established an acute inflammation model and a chronic inflammation model involving adipose stem cells. We used high-throughput miRNA microarray analysis to identify miRNAs that were significantly (p < 0.05) differentially expressed during both acute and chronic inflammation. Lipopolysaccharide (LPS) significantly (p < 0.05) reduced the expression of miR-223-3P and miR-2909, while promoting the production of pro-inflammatory cytokines, interleukin (IL) 6, IL-1ß, and tumor necrosis factor (TNF)-α via the Toll-like receptor (TLR) 4/TLR2/nuclear factor (NF)-κB/signal transducer and activator of transcription (STAT) 3 signaling pathway in human adipose stem cells. Further, miR-223-3P expression was significantly (p < 0.05) reduced in human adipose stem cells during activation by IL-6 stimulation. The inducible down-regulation of miR-223-3P resulted in the activation of STAT3, which was directly targeted by miR-223-3P. STAT3 directly targeted TLR4 and TLR2, promoting the production of the pro-inflammatory cytokine, IL-6, and formed a positive feedback loop to regulate IL-6 levels. Similarly, TNF-α significantly (p < 0.05) increased the expression of miR-223-3p, with LPS and TLR4/TLR2/NF-κB/STAT3 forming a negative feedback loop to regulate TNF-α levels. In addition, miR-2909, which depends on NF-κB, targeted Krueppel-like factor (KLF) 4 to regulate the levels of pro-inflammatory cytokines, IL-6, IL-1ß, and TNF-α. We conclude that miR-223-3p and miR-2909 form a complex regulatory network with pro-inflammatory factors and signaling pathways in adipose stem cells stimulated by LPS. These findings will inform the development of therapies against autoimmune and inflammatory diseases.

Prevalence and Risk Factors of Work-Related Musculoskeletal Disorders Among Intensive Care Unit Nurses in China.

This cross-sectional study investigated the prevalence and risk factors of work-related musculoskeletal disorders among intensive care nurses in the Hunan Province of China. Nurses working in mixed intensive care units of 20 tertiary hospitals in this province participated in an online survey regarding work-related musculoskeletal injuries. The seven-part questionnaire included basic demographics; job and workplace characteristics; risk perception; physical, psychosocial, and workplace organizational factors; and musculoskeletal symptoms. The response rate was 70.7% (702 of 993 nurses). Approximately 97% of the respondents reported experiencing at least one work-related musculoskeletal disorder within the previous year. Low back pain was the most commonly reported musculoskeletal disorder (80.1%), followed by neck (78.6%) and shoulder pain (70.4%). The multivariate logistic regression analysis indicated that work-related musculoskeletal disorders were significantly associated with female gender (odds ratio [OR] = 0.115), unmarried status (OR = 0.136), a greater perception of risk (OR = 2.352), and lack of a safe work environment (OR = 1.056). These findings underscore the need for nurses and managers to reinforce risk awareness, improve physical and psychosocial working conditions, and promote a safer work environment.

Comparative morphology of the leaf epidermis in Lobelia (Lobelioideae) from China.

The leaf epidermal morphology of 38 samples, representing 22 Lobelia species from China, was studied using light microscopy and scanning electron microscopy. The stomata and other epidermal features were largely consistent within species, and therefore represented good characters for taxonomic purposes. Diagnostic characters of the leaf epidermis were used to differentiate species of Lobelia. The species pairs L. clavata and L. pyramidalis, L. alsinoides and L. terminalis, and L. davidii and L. erectiuscula, were differentiated using leaf epidermal characters. Results at the micro-structural level partly supported the current subdivisions of Lobelia. Previous taxonomic revisions of Lobelia in China were evaluated based on species epidermal characteristics. The results supported the recognition of Pratia merged with Lobelia, and of L. chevalieri as a distinct species. Lobelia brevisepala should not be included in L. montana. The status of Lobelia in the evolutionary history of Campanulaceae was assessed. The Lobelia species investigated retained many primitive leaf epidermal features.

[Experimental intervention study of safe injection in basic-level hospitals in Hunan by medical staff].

OBJECTIVE: To experimentally intervene safe injection by medical staff in basic-level hospitals and observe the recent and long-term effect after the intervention and to provide practical measures to improve safe injection. METHODS: We used random sampling methods to set up groups in county hospitals and township hospitals of Hunan Province, and offered lectures, delivered safe injection guide, brochure and on-site guidance in the experimental group. We surveyed the 2 groups after the intervention at 1 month and 6 months to compare the effect of unsafe injection behaviors and safe injection behaviors. RESULTS: One month after the intervention, the unsafe injection rate in the experimental group decreased from 27.8% to 21.7%, while in the control group injection the unsafe injection rate rose from 26.0% to 27.9%, with significant difference (P<0.01). Six months after the intervention, the unsafe injection rate in the experimental group declined to 18.4% while the unsafe injection rate in the control group also dropped to 22.4%, with significant difference (P<0.01). Unsafe injection rate was decreased in the experimental group at different intervention points, with significant difference (P<0.01). The safe injection behavior scores in the experimental group were higher than those in the control group after the intervention of 1 month and 6 month intervention (P<0.01); the experimental group got higher scores after the intervention (P<0.01). CONCLUSION: Training of safe injection, distribution of safe injection guide, and comprehensive intervention model can significantly change the primary care practitioners' behaviors in unsafe injections and it is worth promoting.

catena-Poly[[bis-[2-(2-pyrid-yl)-1-H-imidazole-κN,N]cadmium]-µ-benzene-1,3-dicarboxyl-ato-κO:O].

In the title coordinaltion polymer, [Cd(C(8)H(4)O(4))(C(8)H(7)N(3))(2)](n), the Cd(II) atom, lying on a twofold rotation axis, is six-coordinated by two carboxyl-ate O atoms from two benzene-1,3-dicarboxyl-ate (m-BDC) ligands and four N atoms from two chelating 2-(2-pyrid-yl)imidazole mol-ecules, forming a slightly distorted octa-hedral geometry. The m-BDC ligand is located over a twofold rotation axis. The Cd(II) atoms are bridged by the m-BDC ligands, leading to a wave-shaped chain structure along [010]. N-H⋯O hydrogen bonds connect the chains.

The antitumor and immunostimulating activities of water soluble polysaccharides from Radix Aconiti, Radix Aconiti Lateralis and Radix Aconiti Kusnezoffii.

In the present study, the water-soluble polysaccharides of Radix Aconiti, Radix Aconiti Lateralis and Radix Aconiti Kusnezoffii, were extracted and fractionated into four fractions of each material. The FT-IR and chemical analyses indicated the water-soluble polysaccharides of the three materials were all mainly composed of starch, non-starch type alpha-D-glucans and pectic polysaccharides with different molecular weight distributions and monosaccharide composition ratios. The antitumor assay showed that all the non-starch type polysaccharide fractions had good antitumor activities, and the tumor growth inhibition ratios were 37.24-70.42%. Specifically the inhibition ratios of pectic polysaccharides were over 60%. Moreover, the immunological tests using the Cyclophosphamide (Cy) induced immunosuppressive mice, including phagocytosis of macrophage, NK cell activity, concanavalin A (ConA)-induced T-cell proliferation, lipopolysaccharide (LPS)-induced B-cell proliferation, quantitative haemolysis of sheep red blood cells (SRBC) and dinitro-fluorobenzene (DNFB)-induced delayed-type hypersensitivity (DTH) response assays, exhibited that all the non-starch type polysaccharides, especially the pectic polysaccharide fractions, not only had remarkable immunostimulating activities including nonspecific immunity, cellular immunity and humoral immunity, but also could restore the antitumor drug-suppressed immune function. Therefore, the polysaccharides from Aconitum species might be conveniently exploited to be good immune stimulating modifiers and had the potential to apply in the tumor therapy.

Structure elucidation and antioxidant activity of a novel alpha-(1-->3),(1-->4)-D-glucan from Aconitum kusnezoffii Reichb.

Aconitum kusnezoffii Reichb. has been used as traditional Chinese medicine over the last 2500 years, but its polysaccharides have been paid little attention by now. In the present study, a hot alkali extracted polysaccharide (AKP) from A. kusnezoffii Reichb. was characterized to be an alpha-(1-->3),(1-->4)-D-glucan with Mw 1.4 x 10(5) Da, of which (1-->3)-linked and (1-->4)-linked alpha-Glcp residues were in a ratio of 1:7. In vitro antioxidant testing indicated that AKP had significant ferrous ion-chelating ability, reducing power and scavenging effects on DPPH radical, hydroxyl radical, superoxide anion, H2O2 and self-oxidation of 1,2,3-phentriol, suggesting that it should be explored as a novel natural antioxidant.

The iron-responsive Fur regulon in Yersinia pestis.

The ferric uptake regulator (Fur) is a predominant bacterial regulator controlling the iron assimilation functions in response to iron availability. Our previous microarray analysis on Yersinia pestis defined the iron-Fur modulon. In the present work, we reannotated the iron assimilation genes in Y. pestis, and the resulting genes in complementation with those disclosed by microarray constituted a total of 34 genome loci (putative operons) that represent the potential iron-responsive targets of Fur. The subsequent real-time reverse transcription-PCR (RT-PCR) in conjunction with the primer extension analysis showed that 32 of them were regulated by Fur in response to iron starvation. A previously predicted Fur box sequence was then used to search against the promoter regions of the 34 operons; the homologue of the above box could be predicted in each promoter tested. The subsequent electrophoretic mobility shift assay (EMSA) demonstrated that a purified His(6) tag-fused Fur protein was able to bind in vitro to each of these promoter regions. Therefore, Fur is a global regulator, both an activator and a repressor, and directly controls not only almost all of the iron assimilation functions but also a variety of genes involved in various non-iron functions for governing a complex regulatory cascade in Y. pestis. In addition, real-time RT-PCR, primer extension, EMSA, and DNase I footprinting assay were used to elucidate the Fur regulation of the ybt locus encoding a virulence-required iron uptake system. By combining the published data on the YbtA regulation of ybt, we constructed a concise Fur/YbtA regulatory network with a map of the Fur-promoter DNA interactions within the ybt locus. The data presented here give us an overview of the iron-responsive Fur regulon in Y. pestis.

(E,E)-2,5-Bis(4-tert-butyl-benzyl-idene)-cyclo-penta-none.

The asymmetric unit of the title compound, C(27)H(32)O, contains two and a half mol-ecules. In the crystal structure, one of the mol-ecules lies on a crystallographic twofold rotation axis. The dihedral angles between the benzene rings are 12.17 (6), 16.29 (11) and 51.24 (8)° for the three molecules. The dihedral angles between the benzene rings of each molecule in the asymmetric unit are 12.17 (6) and 16.29 (11)°