Three newOphioceras species (Ascomycetes) from the tropics

Ophioceras guttulatum sp. nov.,O. hongkongense sp. nov. andO. palmae sp. nov. are described and illustrated from decaying terrestrial palms and woody substrates in freshwater habitats. They all have black perithecia with long necks, cylindrical asci with refractive apical rings and filiform ascospores.     

Economic effects and the efficacy of intervention: exploring unintended effects of management and policy on the spread of non-indigenous species

Models have been well developed describing human movements as vectors of the spread of non-indigenous species (NIS). However, to be maximally useful, predictions need to be integrated with management models of how different policies change human behaviour and lead to concurrent changes in invasion risk. Using the dispersal of freshwater organisms by recreational boaters as our study system and mandatory boat washing as our management strategy, we conducted a survey of recreational boaters (n = 580 respondents, t = 2354 boating trips) in Ontario, Canada, and performed counterfactual analysis of boater behavior across different management options. We developed a model to quantify three responses to mandatory boat washing policies: (1) the continued use of a policy lake; (2) switching to a non-policy lake (“trip redistribution”); or, (3) a reduction in boating trips (“trip loss”). We found that boater and locational traits did not have a significant effect, but even modest user fees at washing stations greatly influenced trip redistribution and loss, explaining 87% of the variation in boater choices. These results indicate that user fees can strongly reduce the effectiveness of boat washing programs to mitigate invasion risk and could have unintended local economic effects, supporting the need to minimize boater expense as a program goal. In contrast, only minor redistribution and loss occurred if users washed but did not pay, and when taken together with the lack of effect for boater and locational traits, suggest that simple human-mediated dispersal models would be sufficient to prioritize management actions under “zero fee” scenarios. Simulating management scenarios using an existing spread model for 10 aquatic NIS in Ontario further emphasized the benefit of zero fees. Although averted invasions increased monotonically with effort (number of lakes with washing stations), the relative effectiveness (number of invasions averted per unit effort) was high even with management of a single lake, given zero fees, but required washing stations at far more lakes to maximize relative effectiveness when user fees were imposed.     

Virtual screening and biophysical studies lead to HSP90 inhibitors

Heat shock protein 90 (HSP90) is a molecular chaperone that plays important functional roles in cells. The chaperone activity of HSP90 is regulated by the hydrolysis of ATP at the protein’s N-terminal domain. HSP90, in particular the N-terminal domain, is a current inhibition target for therapeutic treatments of cancers. This paper describes an application of virtual screening, thermal shift assaying and protein NMR spectroscopy leading to the discovery of HSP90 inhibitors that contain the resorcinol structure. The resorcinol scaffold can be found in a class of HSP90 inhibitors that are currently undergoing clinical trials. The proved success of the resorcinol moiety in HSP90 inhibitors validates this combined virtual screen and biophysical technique approach, which may be applied for future inhibitor discovery work for HSP90 as well as other targets.     

Lipopolysaccharide pretreatment increases protease-activated receptor-2 expression and monocyte chemoattractant protein-1 secretion in vascular endothelial cells

Background

This study investigated whether lipopolysaccharide (LPS) increase protease-activated receptor-2 (PAR-2) expression and enhance the association between PAR-2 expression and chemokine production in human vascular endothelial cells (ECs).

Methods

The morphology of ECs was observed through microphotography in cultured human umbilical vein ECs (EA. hy926 cells) treated with various LPS concentrations (0, 0.25, 0.5, 1, and 2 μg/mL) for 24 h, and cell viability was assessed using the MTT assay. Intracellular calcium imaging was performed to assess agonist (trypsin)-induced PAR-2 activity. Western blotting was used to explore the LPS-mediated signal transduction pathway and the expression of PAR-2 and adhesion molecule monocyte chemoattractant protein-1 (MCP-1) in ECs.

Results

Trypsin stimulation increased intracellular calcium release in ECs. The calcium influx was augmented in cells pretreated with a high LPS concentration (1 μg/mL). After 24 h treatment of LPS, no changes in ECs viability or morphology were observed. Western blotting revealed that LPS increased PAR-2 expression and enhanced trypsin-induced extracellular signal-regulated kinase (ERK)/p38 phosphorylation and MCP-1 secretion. However, pretreatment with selective ERK (PD98059), p38 mitogen-activated protein kinase (MAPK) (SB203580) inhibitors, and the selective PAR-2 antagonist (FSLLRY-NH2) blocked the effects of LPS-activated PAR-2 on MCP-1 secretion.

Conclusions

Our findings provide the first evidence that the bacterial endotoxin LPS potentiates calcium mobilization and ERK/p38 MAPK pathway activation and leads to the secretion of the pro-inflammatory chemokine MCP-1 by inducing PAR-2 expression and its associated activity in vascular ECs. Therefore, PAR-2 exerts vascular inflammatory effects and plays an important role in bacterial infection-induced pathological responses.

     

Biomechanical consequences of subtalar joint arthroereisis in treating posterior tibial tendon dysfunction: a theoretical analysis using finite element analysis

Subtalar joint arthroereisis (SJA) has been introduced to control the hyperpronation in cases of flatfoot. The objective of this study is to evaluate the biomechanical consequence of SJA to restore the internal stress and load transfer to the intact state from the attenuated biomechanical condition induced by posterior tibial tendon dysfunction (PTTD). A three-dimensional finite element model of the foot and ankle complex was constructed based on clinical images of a healthy female (age 28 years, height 165 cm, body mass 54 kg). The boundary and loading condition during walking was acquired from the gait experiment of the model subject. Five sets of simulations (conditions) were completed: intact condition, mild PTTD, severe PTTD, mild PTTD with SJA, severe PTTD with SJA. The maximum von Mises stress of the metatarsal shafts and the load transfer along the midfoot during stance were analyzed. Generally, SJA deteriorated the joint force of the medial cuneonavicular and calcaneocuboid joints during late stance, while that of the metatarsocuneiform joints during early stance were over-corrected. Only the calcaneocuboid joint force at 45% stance demonstrated a trend of improvement. Besides, SJA exaggerated the increased stress of the metatarsals compared to the PTTD conditions, except that of the first metatarsal. Our study did not support the hypothesis that SJA can restore the internal load transfer and midfoot stress. SJA cannot compensate the salvage of midfoot stability attributed by PTTD and could be biomechanically insufficient to restore the biomechanical environment. Additional procedures such as orthotic intervention may be necessary.     

Decreased IL-15 may contribute to elevated IgE and acute inflammation in atopic dermatitis.

PBMC and acute skin lesions of patients with atopic dermatitis (AD) are characterized by increased IL-4 and IL-13, but decreased IFN-gamma production. This bias toward an increased Th2 cytokine profile may contribute to the elevated IgE levels and acute skin inflammation seen in AD. In this study, we examined the levels of IL-15, a Th1-like cytokine, in the PBMC and the skin lesions of AD patients. IL-15 secretion by Staphylococcal enterotoxin B-treated PBMC of AD patients was significantly lower than that of normals and psoriasis patients (p < 0.001). Membrane-bound IL-15 expression as measured by mean fluorescence intensity and percentage of IL-15-positive cells in Staphylococcal enterotoxin B-treated monocytes of AD patients (644 +/- 49% and 12.7 +/- 0.6%, respectively) were significantly lower than that of normals (869 +/- 56% and 15.8 +/- 1.2%, respectively) and psoriasis patients (1488 +/- 217% and 22.7 +/- 0.8%, respectively; p < 0.0007 and p < 0.0001, respectively). The membrane-bound IL-15 expression was also significantly lower in the control monocytes of AD patients compared with that in normals and psoriasis patients. There was no significant difference in the absolute number or percentage of monocytes between the study subjects. However, psoriasis skin lesions were found to have significantly more IL-15 mRNA-expressing cells (22.4 +/- 1.7) compared with that in acute AD (7.5 +/- 1.7) and chronic AD (13.7 +/- 1.7) skin lesions (p < 0.05). IL-15 enhanced IFN-gamma production by the PBMC of AD patients (p < 0.01), but not by that of normal individuals or psoriasis patients. In addition, IL-15 was found to suppress IgE synthesis (p < 0.01) by the PBMC of AD patients. These data support the concept that reduced IL-15 expression may contribute to the pathogenesis of AD.     

Mechanism-based partial inactivation of glutathione S-transferases by nitroglycerin: tyrosine nitration vs sulfhydryl oxidation.

Liver glutathione-S-transferases (GSTs) are responsible for the detoxification of electrophiles, and specifically for the metabolism of orally administered organic nitrates such as nitroglycerin (NTG). Recent studies showed that reactive nitrogen species produced by tetranitromethane (TNM), peroxynitrite, or the myeloperoxidase/H2O2/nitrite system can inactivate GST. It is not known whether NTG can similarly inactivate liver GSTs, and if shown, by what mechanism(s). We incubated purified GSTs with NTG, S-nitroso-N-acetylpenicillamine (SNAP), TNM, or vehicle (5% dextrose, D5W), followed by determination of GST activity. Incubation of GST with NTG and TNM caused significant decreases in GST activity whereas no changes were observed with SNAP or D5W. The relative GST activity (vs preincubation) was 73+/-14% for NTG, 37+/-8% for TNM, 98+/-13% for SNAP, and 98+/-9% for D5W, respectively. Exogenous glutathione (GSH) prevented both NTG- and TNM-induced changes in GST activity, consistent with the observed oxidative modification of GST, such as -SH oxidation and dimerization of oxidized GST. In contrast, NTG and TNM exhibited substantial differences in their ability to nitrate tyrosine (TYR) sites in GST. These results demonstrated that NTG can reduce the activity of its own metabolizing enzyme such as GST and this inhibitory effect of NTG was unlikely to be mediated through NO, as such, since SNAP had no effect on GST activity. The partial inactivation of GST by NTG appeared to involve -SH oxidation, but not TYR nitration. These findings provided the first evidence of mechanism-based protein inactivation by NTG, and may lend insight into the hepatic metabolism of NTG and other organic nitrates after repeated oral exposure.     

Synthesis and photophysical properties of new fluorinated benzo[c]xanthene dyes as intracellular pH indicators.

Two new fluorinated benzo[c]xanthene dyes were synthesized by reaction of fluorinated 1,6-dihydroxynaphthalenes with 2,4- (and 2,5)-dicarboxy-3'-dimethylamino-2'-hydroxybenzophenone. The two critical fluorinated 1,6-dihydroxynaphthalene intermediates were prepared via a regioselective route. The fluorinated benzo[c]xanthene dyes exhibit desired lower pK(a) values (6.4 and 7.2, respectively) than their parent compound (pK(a)=7.5) while the pH-dependent dual-emission characteristics are well retained. Their cell-permeable esters have been prepared for intracellular applications.