Pyoluteorin regulates the biosynthesis of 2,4-DAPG through the TetR family transcription factor PhlH in Pseudomonas protegens Pf-5.

Soil and rhizosphere bacteria act as a rich source of secondary metabolites, effectively fighting against a diverse array of pathogens. Certain Pseudomonas species harbor biosynthetic gene clusters for producing both pyoluteorin and 2,4-diacetylphloroglucinol (2,4-DAPG), which are polyketides that exhibit highly similar antimicrobial spectrum against bacteria and fungi or oomycete. A complex cross talk exists between pyoluteorin and 2,4-DAPG biosynthesis, and production of 2,4-DAPG was strongly repressed by pyoluteorin, yet the underlying mechanism is still elusive. In this study, we find that the TetR family transcription factor PhlH is involved in the cross talk between pyoluteorin and 2,4-DAPG biosynthesis. PhlH binds to a palindromic sequence within the promoter of phlG (PphlG), which encodes a C-C bond hydrolase responsible for degrading 2,4-DAPG. As a signaling molecule, pyoluteorin disrupts the PhlH-PphlG complex by binding to PhlH, leading to decreased levels of 2,4-DAPG. Proteomics data suggest that pyoluteorin regulates multiple physiological processes including fatty acid biosynthesis and transportation of taurine, siderophore, and amino acids. Our work not only reveals a novel mechanism of cross talk between pyoluteorin and 2,4-DAPG biosynthesis, but also highlights pyoluteorin's role as a messenger in the complex communication network of Pseudomonas.IMPORTANCEAntibiosis serves as a crucial defense mechanism for microbes against invasive bacteria and resource competition. These bacteria typically orchestrate the production of multiple antibiotics in a coordinated fashion, wherein the synthesis of one antibiotic inhibits the generation of another. This strategic coordination allows the bacterium to focus its resources on producing the most advantageous antibiotic under specific circumstances. However, the underlying mechanisms of distinct antibiotic production in bacterial cells remain largely elusive. In this study, we reveal that the TetR family transcription factor PhlH detects the secondary metabolite pyoluteorin and mediates the cross talk between pyoluteorin and 2,4-DAPG biosynthesis in the biocontrol strain Pseudomonas protegens Pf-5. These findings hold promise for future research, potentially informing the manipulation of these systems to enhance the effectiveness of biocontrol agents.

Combined effects of copper and cadmium exposure on ovarian function and structure in Nile Tilapia (Oreochromis niloticus).

With the rapid development of industrialization and urbanization, the issue of copper (Cu) and cadmium (Cd) pollution in aquatic ecosystems has become increasingly severe, posing threats to the ovarian tissue and reproductive capacity of aquatic organisms. However, the combined effects of Cu and Cd on the ovarian development of fish and other aquatic species remain unclear. In this study, female Nile tilapia (Oreochromis niloticus) were individually or co-exposed to Cu and/or Cd in water. Ovarian and serum samples were collected at 15, 30, 60, 90, and 120 days, and the bioaccumulation, ovarian development, and hormone secretion were analyzed. Results showed that both single and combined exposure significantly reduced the gonadosomatic index and serum hormone levels, upregulated estrogen receptor (er) and progesterone receptor (pr) gene transcription levels, and markedly affected ovarian metabolite levels. Combined exposure led to more adverse effects than single exposure. The data demonstrate that the Cu and Cd exposure can impair ovarian function and structure, with more pronounced adverse effects under Cu and Cd co-exposure. The Cu and Cd affect the metabolic pathways of nucleotides and amino acids, leading to ovarian damage. This study highlights the importance of considering combined toxicant exposure in aquatic toxicology research and provides insights into the potential mechanisms underlying heavy metal-induced reproductive toxicity in fish.

Abscisic acid employs NRP-dependent PIN2 vacuolar degradation to suppress auxin-mediated primary root elongation in Arabidopsis.

How plants balance growth and stress adaptation is a long-standing topic in plant biology. Abscisic acid (ABA) induces the expression of the stress-responsive Asparagine Rich Protein (NRP), which promotes the vacuolar degradation of PP6 phosphatase FyPP3, releasing ABI5 transcription factor to initiate transcription. Whether NRP is required for growth remains unknown. We generated an nrp1 nrp2 double mutant, which had a dwarf phenotype that can be rescued by inhibiting auxin transport. Insufficient auxin in the transition zone and over-accumulation of auxin at the root tip was responsible for the short elongation zone and short-root phenotype of nrp1 nrp2. The auxin efflux carrier PIN2 over-accumulated in nrp1 nrp2 and became de-polarized at the plasma membrane, leading to slower root basipetal auxin transport. Knock-out of PIN2 suppressed the dwarf phenotype of nrp1 nrp2. Furthermore, ABA can induce NRP-dependent vacuolar degradation of PIN2 to inhibit primary root elongation. FyPP3 also is required for NRP-mediated PIN2 turnover. In summary, in growth condition, NRP promotes PIN2 vacuolar degradation to help maintain PIN2 protein concentration and polarity, facilitating the establishment of the elongation zone and primary root elongation. When stressed, ABA employs this pathway to inhibit root elongation for stress adaptation.

RNA-mediated gene fusion in mammalian cells.

One of the hallmarks of cancer is the formation of oncogenic fusion genes as a result of chromosomal translocations. Fusion genes are presumed to form before fusion RNA expression. However, studies have reported the presence of fusion RNAs in individuals who were negative for chromosomal translocations. These observations give rise to "the cart before the horse" hypothesis, in which the genesis of a fusion RNA precedes the fusion gene. The fusion RNA then guides the genomic rearrangements that ultimately result in a gene fusion. However, RNA-mediated genomic rearrangements in mammalian cells have never been demonstrated. Here we provide evidence that expression of a chimeric RNA drives formation of a specified gene fusion via genomic rearrangement in mammalian cells. The process is: (i) specified by the sequence of chimeric RNA involved, (ii) facilitated by physiological hormone levels, (iii) permissible regardless of intrachromosomal (TMPRSS2-ERG) or interchromosomal (TMPRSS2-ETV1) fusion, and (iv) can occur in normal cells before malignant transformation. We demonstrate that, contrary to "the cart before the horse" model, it is the antisense rather than sense chimeric RNAs that effectively drive gene fusion, and that this disparity can be explained by transcriptional conflict. Furthermore, we identified an endogenous RNA AZI1 that functions as the "initiator" RNA to induce TMPRSS2-ERG fusion. RNA-driven gene fusion demonstrated in this report provides important insight in early disease mechanisms, and could have fundamental implications in the biology of mammalian genome stability, as well as gene-editing technology via mechanisms native to mammalian cells.

Recurrent BCAM-AKT2 fusion gene leads to a constitutively activated AKT2 fusion kinase in high-grade serous ovarian carcinoma.

High-grade serous ovarian cancer (HGSC) is among the most lethal forms of cancer in women. Excessive genomic rearrangements, which are expected to create fusion oncogenes, are the hallmark of this cancer. Here we report a cancer-specific gene fusion between BCAM, a membrane adhesion molecule, and AKT2, a key kinase in the PI3K signaling pathway. This fusion is present in 7% of the 60 patient cancers tested, a significant frequency considering the highly heterogeneous nature of this malignancy. Further, we provide direct evidence that BCAM-AKT2 is translated into an in-frame fusion protein in the patient's tumor. The resulting AKT2 fusion kinase is membrane-associated, constitutively phosphorylated, and activated as a functional kinase in cells. Unlike endogenous AKT2, whose activity is tightly regulated by external stimuli, BCAM-AKT2 escapes the regulation from external stimuli. Moreover, a BCAM-AKT2 fusion gene generated via chromosomal translocation using the CRISPR/Cas9 system leads to focus formation in both OVCAR8 and HEK-293T cell lines, suggesting that BCAM-AKT2 is oncogenic. Together, the results indicate that BCAM-AKT2 expression is a new mechanism of AKT2 kinase activation in HGSC. BCAM-AKT2 is the only fusion gene in HGSC that is proven to translate an aberrant yet functional kinase fusion protein with oncogenic properties. This recurrent genomic alteration is a potential therapeutic target and marker of a clinically relevant subtype for tailored therapy of HGSC.

Preparation of Graphite Oxide Containing Different Oxygen-Containing Functional Groups and the Study of Ammonia Gas Sensitivity.

A series of graphite oxide samples were prepared using the modified Hummers method. Flake graphite was used as the raw material and the reaction temperature of the aqueous solution was changed (0 °C, 30 °C, 50 °C, 60 °C, 70 °C, 80 °C, and 100 °C). X-ray diffraction, Fourier-transform infrared spectroscopy, Raman spectral analysis, X-ray photoelectron spectroscopy, and contact angle tests were performed to characterize the structure, chemical bonding, type, and content of oxygen-containing functional groups of the graphite oxide samples. The results showed that the type and content of each oxygen-containing functional group could be controlled by changing the reaction temperature with the addition of water. As the temperature of the system increased, the degree of oxidation of the graphite oxide samples first increased and then decreased. Too high a temperature (100 °C) of the system led to the formation of epoxy groups by the decomposition of some hydroxyl groups in the samples, causing the reduction of oxygen-containing functional groups between the graphite layers, poor hydrophilic properties, and low moisture content. When the system temperature was 50 °C, the interlayer spacing of the graphite oxide samples was at its highest, the graphite was completely oxidized (C/O = 1.85), and the oxygen-containing functional groups were mainly composed of hydroxyl groups (accounting for approximately 28.88% of the total oxygen-containing functional groups). The high content of hydroxyl and carboxyl groups had good hydrophilic ability and showed the highest moisture content. The sample at 50 °C had better sensitivity to ammonia because of its high hydroxyl group and carboxyl group content, with the sample showing an excellent profile when the ammonia concentration was 20⁻60 ppm.

The in vitro selection world.

Through iterative cycles of selection, amplification, and mutagenesis, in vitro selection provides the ability to isolate molecules of desired properties and function from large pools (libraries) of random molecules with as many as 10(16) distinct species. This review, in recognition of a quarter of century of scientific discoveries made through in vitro selection, starts with a brief overview of the method and its history. It further covers recent developments in in vitro selection with a focus on tools that enhance the capabilities of in vitro selection and its expansion from being purely a nucleic acids selection to that of polypeptides and proteins. In addition, we cover how next generation sequencing and modern biological computational tools are being used to complement in vitro selection experiments. On the very least, sequencing and computational tools can translate the large volume of information associated with in vitro selection experiments to manageable, analyzable, and exploitable information. Finally, in vivo selection is briefly compared and contrasted to in vitro selection to highlight the unique capabilities of each method.

Land management influences trade-offs and the total supply of ecosystem services in alpine grassland in Tibet, China.

Developing sustainable use patterns for alpine grassland in Tibet is the primary challenge related to conserving these vulnerable ecosystems of the 'world's third pole' and guaranteeing the well-being of local inhabitants. This challenge requires researchers to think beyond the methods of most current studies that are limited to a single aspect of conservation or productivity, and focus on balancing various needs. An analysis of trade-offs involving ecosystem services provides a framework that can be used to quantify the type of balancing needed. In this study, we measured variations in four types of ecosystem services under five types of grassland management including grazing exclusion, sowing, combined plowing and grazing exclusion, combined plowing and sowing, and natural grassland, from 2013 to 2015. In addition, we accessed the existence and changing patterns of ecosystem service trade-offs using Spearman coefficients and a trade-off index. The results revealed the existence of trade-offs among provisioning and regulating services. Plowing and sowing could convert the trade-off relationships into synergies immediately. Grazing exclusion reduced the level of trade-offs gradually over time. Thus, the combined plowing and sowing treatment promoted the total supply of multiple ecosystem services when compared with natural grassland. We argue that the variations in dry matter allocation to above- and belowground serve as one cause of the variation in trade-off relationships. Another cause for variation in trade-offs is the varied species competition between selection effects and niche complementarity. Our study provides empirical evidence that the effects of trade-offs among ecosystem services could be reduced and even converted into synergies by optimizing management techniques.

Water Injection Into Coal Seams for Outburst Prevention: The Coupling Effect of Gas Displacement and Desorption Inhibition.

Gassy coal seams generally have low permeability and dry coal bodies, which are susceptible to coal and gas outburst hazards in the process of mining. Water injection into coal seams can significantly alter the gas release rate and flow behavior. However, water has dual effects on coal seams: gas displacement and water-locking, and the coupling mechanism of these two effects is not clear in the whole process of coal seam water injection. By measuring high-pressure gas adsorption isotherms and gas diffusion initial velocity, it was found that both the Langmuir adsorption constant a and gas diffusion initial velocity ΔP decrease with the increase of water content, which would reduce outburst risks. Through the self-developed integrated experimental device of "gas adsorption + water injection displacement + gas desorption″, the changing rules of gas displacement amount, desorption amount, and water lock amount under different water injection conditions were studied. The results show that when the water injection ratio increases from 6 to 25%, the gas displacement would increase from 0.62 to 1.16 mL/g, with an increase of 87.09%. Also, at the same time, the gas desorption capacity would decrease from 4.86 to 4.05 mL/g after pressure relief, with a decrease of 16.67%. The amount of water-locking increased from 0.11 to 0.38 mL/g. The effect of water injection to control coal and gas outburst occurs in two different water injection stages. In the process of water injection, water plays a major role in gas displacement, which is conducive to reducing the gas content of the coal seam. After the completion of water injection, the static water pressure remaining in the coal seam can reduce the gas emission rate. The combination of these two effects can effectively reduce the risk of outbursts.

Ginger volatile oil inhibits the growth of MDA-MB-231 in the bisphenol A environment by altering gut microbial diversity.

To examine the impact of ginger volatile oil (GVO) on the growth of MDA-MB-231 breast cancer cells in the presence of bisphenol A (BPA) by modulating the diversity of gut microbiota. METHODS: MDA-MB-231 breast cancer cells were injected subcutaneously into the right armpit of female BALB/c Nude (nu/nu) mice to create a triple negative breast cancer model. Thirty nude mice were randomly divided into 5 groups: control group (distilled water every day), BPA control group (distilled PEG-400+ DMSO + cyclodextrin every day), BPA + GVO (0.25 mL/kg) group, BPA + GVO (0.5 mL/kg) group, BPA + GVO (1 mL/kg) group, 6 mice in each group; The drug was given by gavage once a day for 4 weeks. At the end of the experiment, the changes of tumor mass and tumor volume were observed and compared in 5 groups of tumor-bearing mice. High-throughput sequencing (16S rRNA) was used to detect the changes of gut microflora in each group. RESULTS: The volume and weight of breast cancer decreased in the low, medium and high dose groups of GVO. Among them, the difference between the high-dose group and the BPA group reached a significant level (P < 0.05). The species and abundance of gut flora decreased following BPA treatment, but increased after combined treatment of BPA with GVO. In the tumor control group, the ratio of Firmicutes(F) and Bacteroidea(B) respectively was 0.10:0.79 at the phylum level, while the ratio of BPA group further decreased (0.04:0.88). After feeding GVO, the number of Firmicutes and Bacteroidea increased, the F/B ratio increased, and the level of Lactobacillus and alistipes increased. In the BPA and GVO treatment group, the predominant gut microflora functions are cell membrane biogenesis, carbohydrate transport and metabolism. This is followed by amino acid transport and metabolism, and transcription function. After GVO administration, the Gram-positive bacteria (G+) ratio had an increasing trend and the Gram-negative bacteria (G-)ratio had a decreasing trend. CONCLUSION: The species and abundance of gut flora decreased following BPA treatment, but increased after combined treatment of BPA with GVO.

Control of mammalian gene expression by modulation of polyA signal cleavage at 5' UTR.

The ability to control gene expression in mammalian cells is crucial for safe and efficacious gene therapies and for elucidating gene functions. Current gene regulation systems have limitations such as harmful immune responses or low efficiency. We describe the pA regulator, an RNA-based switch that controls mammalian gene expression through modulation of a synthetic polyA signal (PAS) cleavage introduced into the 5' UTR of a transgene. The cleavage is modulated by a 'dual-mechanism'-(1) aptamer clamping to inhibit PAS cleavage and (2) drug-induced alternative splicing that removes the PAS, both activated by drug binding. This RNA-based methodology circumvents the immune responses observed in other systems and achieves a 900-fold induction with an EC50 of 0.5 µg ml-1 tetracycline (Tc), which is well within the FDA-approved dose range. The pA regulator effectively controls the luciferase transgene in live mice and the endogenous CD133 gene in human cells, in a dose-dependent and reversible manner with long-term stability.

[Application of throat swab nested PCR in the diagnosis of congenital human cytomegalovirus infection in neonates].

OBJECTIVE: To evaluate the clinical application value of throat swab nested PCR for detecting active congenital human cytomegalovirus (HCMV) infection in neonates. METHODS: The throat swabs and umbilical cord blood specimens from 51 neonates were collected for nested PCR assay for HCMV glycoprotein B (gB) gene. Moreover, 18 of them were subjected to a pp65 antigen test. RESULTS: The sensitivity and specificity of throat swab nested PCR for HCMV gB gene were 67% and 75%, respectively, and the positive and negative predictive values were 57% and 82%, respectively. CONCLUSIONS: Throat swab nested PCR assay for HCMV gB gene is non-invasive, rapid, and highly sensitive for HCMV detection and holds promise as an excellent screening technology for detecting active congenital HCMV infection in neonates.

Influencing Mechanism of Titanium-Extracted Tailing Slag on the Strength of CaO Steel Slag Hardened Paste.

Hardened pastes with different mass percentages of steel slag (SS)/titanium-extracted tailing slag (TETS) were prepared under fixed CaO content to determine the influencing mechanism of TETS on the strength of CaO SS hardened paste. Furthermore, the effects and laws of curing time and SS/TETS ratios on the strength of hardened pastes were also investigated in this study. Importantly, hydration products, microstructures and the micro-area compositions of hardened pastes were analysed using X-ray diffraction, Fourier-transform infrared spectroscopy and scanning electron microscopy-energy dispersive spectrometer, respectively, to reveal the influencing mechanism of TETS on the CaO SS hardened pastes. The results demonstrated that the early strength of hardened pastes increases considerably following the inclusion of TETS. Specifically, the strength of the sample with an SS/TETS ratio of 22.5:67.5 at 1 d can be increased by more than 14 times. Notably, its strength at 90 days reached 19.36 MPa. Moreover, the diffraction peaks of calcite and C-S-H in the samples were also strengthened. Meanwhile, a diffraction peak of hydrocalumite appeared, and the calcites in the samples were curled up. When the SS/TETS ratio was equal to or more than 45:45, a diffraction peak of Ca(OH)2 appeared in the sample. Only a diffraction peak of Ca(OH)2 and weak diffraction peaks of calcite and C-S-H were observed in the samples without TETS, but there was no diffraction peak of hydrocalumite. The strength at 90 days was only 4.92 MPa. The increased strength of the hardened paste is closely related to the production of new phases after adding TETS. Solid particles in the hardened paste are cemented into a whole because of the hydration of C-S-H. Calcite forms the skeleton of the hardened pastes, whereas hydrocalumite fills in the pores among particles in hardened pastes, thus making them more compacted. As a result, there is increased.

Exploring the mechanism of an active ingredient of ginger, dihydrocapsaicin, on triple negative breast cancer based on network pharmacology and in vitro experiments.

To investigate the potential mechanism of ginger in the treatment of triple-negative breast cancer (TNBC) based on network pharmacology, molecular docking and in vitro cell experiments. The Traditional Chinese Medicine Systems Pharmacology Database And Analysis Platform, the Bioinformatics Analysis Tool For Molecular Mechanism Of Traditional Chinese Medicine and the HERB database and literature search were used to search for the main active compounds of ginger. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were used to predict the possible molecular mechanism and signaling pathway of ginger in the treatment of triple negative breast cancer. The key core genes of ginger in the treatment of triple negative breast cancer were docked with the active ingredients of ginger on the Autodock platform, and the mechanism of ginger on triple negative breast cancer was further verified by in vitro cell experiments. As a result, 10 effective components, 27 potential targets and 10 Protein-Protein Interaction core genes were predicted in the treatment of triple negative breast cancer with ginger, involving 287 biological processes, 18 cellular components and 38 molecular functions. Ginger regulated the proliferation, migration and apoptosis of triple negative breast cancer cells by regulating TNF, IL-17, FoxO, MAPK, PI3K/AKT and other signaling pathways. The results of molecular docking showed that the lowest binding potential energy between dihydrocapsaicin (DHC) and EGFR protein was -7.70 kcal·mol-1, followed by that between 6-gingerol and EGFR protein was -7.30 kcal·mol-1 and that between DHC and CASP3 protein was -7.20 kcal·mol-1. In vitro cell experiments showed that ginger could inhibit the proliferation and migration of TNBC MDA-MB-231 cells, increase the mRNA expression of Caspase family CASP9 and the protein expression of CASP3 and BAX. Overall, based on the combination of network pharmacology and in vitro cell experiments, ginger has the characteristics of multi-target in the treatment of TNBC, which may play a regulatory role through the PI3K/AKT family. It provides a reference for the drug development of ginger and the clinical treatment of triple negative breast cancer.

Influences of Friedel's Salt Produced by CaO-Activated Titanium-Extracted Tailing Slag on Chloride Binding.

Titanium-extracted tailing slag (TETS) has high activity, but the content of chloride ions is high. To effectively bind the chloride ions, CaO was used to activate the TETS, and the solidified cementitious material of CaO-activated TETS was prepared. The effects of CaO content and curing age on the strength of solidified samples, chloride binding capacity, and chloride binding mechanism were studied. By means of XRD, FTIR, SEM, and EDS, the hydration reaction products, microstructure, morphology, and micro-components of the solidified sample were characterized. The results show that the chloride ions can be effectively bound by using CaO to activate TETS with higher mechanical strength. When the CaO content is 10 wt%, the strength of the 28-day-cured body can reach more than 20 MPa, the chloride ion binding amount is 38.93 mg/g, and the chloride binding rate is as high as 68%. The new product phases of the solidified sample are mainly Friedel's salt (FS) and calcite, in which the amount of FS production and the degree of crystal development are affected by the CaO content and curing age. The chloride binding ions in the solidified sample are mainly the chemical binding by FS. The FS diffraction peak strength increases with the increase of CaO content and curing age, but the calcite diffraction peak strength is less affected by them. FS mainly accumulates and grows in the pores of the solidified sample. It can optimize the pore structure of the solidified sample and improve the strength of the solidified sample while binding chloride ions. The results can provide useful information for the resource utilization of chlorine-containing TETS, the improvement of durability of Marine concrete, and the application of sea sand in concrete.

Dietary ginger polysaccharides (Gps) improve symptoms in hyperlipidemia rats via alterations in gut microbiota.

The aim of this research is to investigate lipid-lowering influence of dietary ginger (Zingier officinales Rocs) polysaccharides (GPS) on hyperlipidemia rats. Rat models with hyperlipidemia was established by high-fat food diet (HFD). Comparing to GP-negative model group, GPS attenuated several effects of HFD feeding, including the levels of blood lipid biochemistry, serum inflammatory markers (tumor necrosis factor TNF-a, interleukin IL-6), antioxidant capacity (superoxide dismutase SOD, glutathione peroxidase GSH-Px, total antioxidant capacity T-AOC, propylene dialdehyde MDA), uric acid and immune index. 16 S rDNA gene sequencing of fecal samples showed that GPS increased the growth of Akkermansia muciniphila and decreased the proportion of Firmicutes to Bacteroidetes; This changes in microbial community structure can help prevent diet-induced metabolic disease. These results suggest that GPs may act on the gut, changing the structure of the gut microbial community, thereby reducing intestinal and systemic inflammation, thus improved metabolic outcomes.

Comparison of magnetoimpedance behaviors in meander CoFeNiSiB amorphous ribbon for deformation sensing applications.

In this work, micro-ribbon strips and meanders based on CoFeNiSiB amorphous ribbons were fabricated by using the lithography technique and chemical etching. Flat and curved holders with different radius of curvature were obtained via 3D printing techniques for GMI testing. Longitudinal and transverse GMI (LGMI and TGMI) behaviors of micro-ribbon sensors in different bending directions and degrees were systematically investigated. The results show the LGMI and TGMI effects of micro-ribbon meanders with one turn is most sensitive to bending. It can be used in the development of deformation sensors. In addition, there is a linear range of field in the LGMI and TGMI curves of micro ribbons under different bending conditions, and the sensitivity of micro-ribbon sensors shows no significant change in the range. In particular, the micro-ribbon meanders with three turns are the least sensitive to bending deformation and can be used to develop stable and flexible GMI sensors for wearable electronics devices.

Effects of decoction of Angelica Sinensis, Zingiberis Rhizoma Recens, and mutton on physiology and biochemistry of Sprague Dawley female rats with spleen-kidney Yang deficiency.

Background: To examine the effects of each dose of decoction of Angelica sinensis (Dang gui), Zingiberis Rhizoma Recens (Sheng jiang), and mutton (DAZM) on the physiological and biochemical indexes of female rats with spleen-kidney Yang deficiency (SKYD) through 30-day feeding of DAZM, and to evaluate the tonifying effect of DAZM combined with the system of benefit damage index-general score (BDI-GS). Methods: Sprague Dawley (SD) rats were administered adenine and senna water to establish a SKYD model. The rats were then allocated to 4 groups at random: Model group, and L group, 4.2 g/kg, M group, 8.4 g/kg and H group, 16.8 g/kg. In addition, the group of normal feeding with unlimited diet was set as N group. Blood samples were taken to detect the relevant physiological and biochemical indexes. For organ coefficient analysis, 10 kinds of organ tissues were dissected and weighed. The tonifying effect of DAZM was discussed according to the BDI-GS system. Results: During the modeling, the weight of rats in the normal group displayed a marked growth trend, and the weight of the model group was markedly lower than that of the normal group. After feeding the rats DAZM at a low, intermediate, and high dose, the anal temperature of rats in each group continued to rise, and finally remained basically the same as that of normal rats. Hematological and urine examinations revealed that the urea nitrogen and creatinine (CRE) of the model group and the experimental group were markedly higher than those of the normal group, and there were marked differences. After intragastric administration of DAZM, E2 increased markedly. The BDI-GS values of the liver, spleen, lung, kidney, brain, ovary, and adrenal gland of female rats in the 3 administration groups of DAZM were >1, and the total cumulative GS value of each organ of female rats was more than 10. Conclusions: The decoction of DAZM has no obvious effect on the growth, metabolism, and development of female rats with SKYD, causes no obvious damage to organs, and has a certain reparative effect on the kidney damage caused by SKYD.

Effects of four hormones on the mitigation of ovarian damage in tilapia (Oreochromis niloticus) after copper and cadmium exposure.

Female tilapia of the Genetic Improvement of Farmed Tilapia (GIFT) strain were selected as an animal model to study the effects of four hormonal drugs in mitigating ovarian damage following exposure to copper and cadmium. After combined exposure to copper and cadmium in aqueous phase for 30 d, tilapia were randomly injected with oestradiol (E2), human chorionic gonadotropin (HCG), luteinizing hormone releasing hormone (LHRH), or coumestrol and raised in clear water for 7 d Ovarian samples were collected after combined exposure to heavy metals for 30 d and after recovery for 7 d Gonadosomatic index (GSI), copper and cadmium levels in the ovary, reproductive hormone levels in serum, and mRNA expression of key reproductive regulatory factors were determined. After 30 d of exposure to the combined copper and cadmium in aqueous phase, the Cd2+ content in tilapia ovarian tissue increased by 1,242.46% (p < 0.05), whereas the Cu2+ content, body weight, and GSI decreased by 68.48%, 34.46%, and 60.00% (p < 0.05), respectively. Additionally, E2 hormone levels in tilapia serum decreased by 17.55% (p < 0.05). After drug injection and recovery for 7 d, compared to the negative control group, the HCG group exhibited an increase of 39.57% (p < 0.05) in serum vitellogenin levels. Increases of 49.31%, 42.39%, and 45.91% (p < 0.05) in serum E2 levels were observed, and mRNA expression of 3ß-HSD increased by 100.64%, 113.16%, and 81.53% (p < 0.05) in the HCG, LHRH, and E2 groups, respectively. The mRNA expression of CYP11A1 in tilapia ovaries increased by 282.26% and 255.08% (p < 0.05) and mRNA expression of 17ß-HSD increased by 109.35% and 111.63% in the HCG and LHRH groups, respectively (p < 0.05). All four hormonal drugs, particularly HCG and LHRH, promoted the restoration of tilapia ovarian function to varying degrees after injury induced by combined exposure to copper and cadmium. This study presents the first hormonal treatment protocol for the mitigation of ovarian damage in fish exposed to combined aqueous phases of copper and cadmium as a strategy to prevent and treat fish ovarian damage induced by heavy metals.

A novel rhodanese is required to maintain chloroplast translation in Chlamydomonas.

Rhodanese-domain proteins (RDPs) are widespread in plants and other organisms, but their biological roles are mostly unknown. Here we report on a novel RDP from Chlamydomonas that has a single rhodanese domain, and a predicted chloroplast transit peptide. The protein was produced in Escherichia coli with a His-tag, but lacking most of the N-terminal transit peptide, and after purification was found to have rhodanese activity in vitro. It was also used to elicit antibodies for western blot analysis, which showed that the native Chlamydomonas protein migrated slower on SDS gels (apparent M(r) =34 kDa) than its predicted size (27 kDa), and co-fractionated with chloroplasts. To assess function in vivo, the tandem-RNAi approach was used to generate Chlamydomonas strains that had reductions of 30-70% for the mRNA and ~20-40% for the 34-kDa protein. These strains showed reduced growth under all trophic conditions, and were sensitive to even moderate light; properties reminiscent of chloroplast translation mutants. Pulse-labeling in the presence of cycloheximide indicated that chloroplast protein synthesis was broadly reduced in the RNAi strains, and transcript analysis (by RT-PCR and northern blotting) indicated the effect was mainly translational. These results identify a novel rhodanese-like protein that we have named CRLT, because it is required to maintain chloroplast translation.