Chemoproteomic strategy identifies PfUCHL3 as the target of halofuginone.

The human malaria parasite Plasmodium falciparum (P. falciparum) continues to pose a significant public health challenge, leading to millions of fatalities globally. Halofuginone (HF) has shown a significant anti-P. falciparum effect, suggesting its potential as a therapeutic agent for malaria treatment. In this study, we synthesized a photoaffinity labeling probe of HF to identify its direct target in P. falciparum. Our results reveal that ubiquitin carboxyl-terminal hydrolase 3 (PfUCHL3) acts as a crucial target protein of HF, which modulates parasite growth in the intraerythrocytic cycle. In particular, we discovered that HF potentially forms hydrogen bonds with the Leu10, Glu11, and Arg217 sites of PfUCHL3, thereby inducing an allosteric effect by promoting the embedding of the helix 6' region on the protein surface. Furthermore, HF disrupts the expression of multiple functional proteins mediated by PfUCHL3, specifically those that play crucial roles in amino acid biosynthesis and metabolism in P. falciparum. Taken together, this study highlights PfUCHL3 as a previously undisclosed druggable target of HF, which contributes to the development of novel anti-malarial agents in the future.

[Arachidonic acid Alox15/12-HETE signaling inhibits vascular calcification].

This study aims to explore the effects of arachidonic acid lipoxygenase metabolism in vascular calcification. We used 5/6 nephrectomy and high-phosphorus feeding to establish a model of vascular calcification in mice. Six weeks after nephrectomy surgery, vascular calcium content was measured, and Alizarin Red S and Von Kossa staining were applied to detect calcium deposition in aortic arch. Control aortas and calcified aortas were collected for mass spectrometry detection of arachidonic acid metabolites, and active molecules in lipoxygenase pathway were analyzed. Real-time quantitative PCR was used to detect changes in the expression of lipoxygenase in calcified aortas. Lipoxygenase inhibitor was used to clarify the effect of lipoxygenase metabolic pathways on vascular calcification. The results showed that 6 weeks after nephrectomy surgery, the aortic calcium content of the surgery group was significantly higher than that of the sham group (P < 0.05). Alizarin Red S staining and Von Kossa staining showed obvious calcium deposition in aortic arch from surgery group, indicating formation of vascular calcification. Nine arachidonic acid lipoxygenase metabolites were quantitated using liquid chromatography/mass spectrometry (LC-MS) analysis. The content of multiple metabolites (12-HETE, 11-HETE, 15-HETE, etc.) was significantly increased in calcified aortas, and the most abundant and up-regulated metabolite was 12-HETE. Furthermore, we examined the mRNA levels of metabolic enzymes that produce 12-HETE in calcified blood vessels and found the expression of arachidonate lipoxygenase-15 (Alox15) was increased. Blocking Alox15/12-HETE by Alox15 specific inhibitor PD146176 significantly decreased the plasma 12-HETE content, promoted calcium deposition in aortic arch and increased vascular calcium content. These results suggest that the metabolism of arachidonic acid lipoxygenase is activated in calcified aorta, and the Alox15/12-HETE signaling pathway may play a protective role in vascular calcification.

Mechanisms of potentially toxic metal removal from biogas residues via vermicomposting revealed by synchrotron radiation-based spectromicroscopies.

Biogas residues (BR) contaminated with potentially toxic metals pose environmental risks to soils and food chains, and strategies are needed to decrease the concentration and bioavailability of potentially toxic metals in BR. Here, metal fractions and removal mechanisms were quantified by synchrotron radiation-based Fourier transform infrared and micro X-ray fluorescence spectromicroscopies on BR and earthworms subject to vermicomposting. Vermicomposting resulted in decreases in concentrations of potentially toxic metals in BR and increases in metal removal efficiencies due to uptake by earthworms. Prior to vermicomposting, Zn, Cu and Pb were associated with N-H, O-H, aromatic C, aliphatic C, and amide functional groups, but following maturation during vermicomposting, metals were associated with N-H, O-H, aliphatic C and polysaccharide functional groups. Following vermicomposting, Zn and Cu were mainly distributed in the dermal portions of earthworms, whereas Pb was more homogeneously distributed among the inner and outer portions of the earthworms, revealing that different metals may have different uptake routes. These findings provide a new strategy for safe utilization of BR by using earthworms via vermicomposting to remove potentially toxic metals and in situ insights into how metals binding and distribution characteristics in BR and earthworms during compost and vermicomposting processes.

Cardiac Tamponade as Initial Presentation of Angioimmunoblastic T-Cell Lymphoma.

A 21-year-old man presented initially with impending cardiac tamponade secondary to an angioimmunoblastic T-cell lymphoma (AITL). Following unsuccessful pericardiocentesis and subxiphoid pericardiostomy, the patient's deteriorating hemodynamics prompted an urgent sternotomy. Histopathological diagnosis confirmed AITL. While chemotherapy remains the most effective treatment of AITL, life-threatening complications of the tumor necessitates surgical intervention.

Cloning and characterization of a salt responsive gene AcPsbQ1 from Atriplex canescens.

PsbQ is an extrinsic subunit of the photosystem II in eukaryotic photosynthetic organisms. Numerous studies have demonstrated that PsbQ can stabilize the inorganic cofactors and enhance the oxygen release in PSII. The decrease of photosynthesis rate under salinity condition is normally attributed to the high concentration of injurious ions, such as Na(+) and Cl(-), which accumulate in the chloroplast and damage thylakoid membrane under salinity stress. In this study, AcPsbQ1 was isolated from a halophyte Atriplex canescens cDNA library. The AcPsbQ1 contains an open reading frame of 699 bp encoding a 233 amino acid protein. In order to investigate its function, AcPsbQ1 was cloned and transformed into Saccharomyces cerevisiae INVSc1. The heterologous expression of AcPsbQ1 in transgenic yeast significantly helped to increase the adapting and recovery ability of yeast cells under the salt and drought. Quantitative real-time PCR assay was performed to reveal the expression pattern of AcPsbQ1 under different abiotic stresses. On exposure to NaCl stress, the transcript level of AcPsbQ1 was significantly enhanced. AcPsbQ1 expression level was also up-regulated under drought stress. These results indicated that AcPsbQ1 might involve in the response to salt stress in A. canescens.

A heavy metal-associated protein (AcHMA1) from the halophyte, Atriplex canescens (Pursh) Nutt., confers tolerance to iron and other abiotic stresses when expressed in Saccharomyces cerevisiae.

Many heavy metals are essential for metabolic processes, but are toxic at elevated levels. Metal tolerance proteins provide resistance to this toxicity. In this study, we identified and characterized a heavy metal-associated protein, AcHMA1, from the halophyte, Atriplex canescens. Sequence analysis has revealed that AcHMA1 contains two heavy metal binding domains. Treatments with metals (Fe, Cu, Ni, Cd or Pb), PEG6000 and NaHCO3 highly induced AcHMA1 expression in A. canescens, whereas NaCl and low temperature decreased its expression. The role of AcHMA1 in metal stress tolerance was examined using a yeast expression system. Expression of the AcHMA1 gene significantly increased the ability of yeast cells to adapt to and recover from exposure to excess iron. AcHMA1 expression also provided salt, alkaline, osmotic and oxidant stress tolerance in yeast cells. Finally, subcellular localization of an AcHMA1/GFP fusion protein expressed in tobacco cells showed that AcHMA1 was localized in the plasma membrane. Thus, our results suggest that AcHMA1 encodes a membrane-localized metal tolerance protein that mediates the detoxification of iron in eukaryotes. Furthermore, AcHMA1 also participates in the response to abiotic stress.

[High accuracy sample positioning system for hard X-ray microprobe].

A novel sample offline positioning system was developed for hard X-ray micro-focus beamline (BL15U1) at Shanghai Synchrotron Radiation Facility (SSRF). The positioning system is composed of three parts: off-line sample microscope system, on-line sample experiment system, and high-precision positioning sample holder. It makes a potent combination of the on-line X-ray fluorescence imaging and the off-line microscopic examination in three steps: compiling of control program, positioning of sample holder, and conversion of the two coordinates. It's the first time in the domestic synchrotron radiation facilities to achieve sample offline positioning in micron scale. The system helps the researchers find the object of study in micro zone quickly and accurately, when they study the micro characteristics of materials using synchrotron radiation micro X-ray beam. The gold mesh was used as an object of study. By comparing the differences of coordinates of gold mesh nodes between pictures from offline microscope and pictures from X-ray fluorescence mapping, the accuracy of the offline positioning system was verified. The results showed that the average errors of X-axis and Z-axis were 1.3 and 2.5 microm respectively, using the positioning method. It was demonstrated that the sample offline positioning system not only is suitable for these applications with high efficiency, but also supply hard X-ray micro-focus beamline with the technical preparations of sample automatic focusing method.