Mitochonic Acid 5 Increases Ram Sperm Quality by Improving Mitochondrial Function during Storage at 4 °C.

Semen preservation involves lengthening sperm's fertile lifespan without any detrimental effects on its biochemical, functional, and ultrastructural properties. Liquid storage at 4 °C is a ram sperm preservation method. However, this method of storage causes irreversible damage due to cold shocks, osmotic stresses, oxidative stresses, and reductions in sperm metabolism. The present study aims to investigate whether the supplementation of mitochonic acid 5 (MA-5) in a sperm extender could improve chilled ram sperm quality and elucidate its mechanism of action. Ram sperm were diluted with a tris-citrate-glucose extender containing different concentrations of MA-5 (0, 0.1, 1, 10, and 100 nM) and stored at 4 °C for up to 48 h. Sperm motility, membrane integrity, acrosome integrity, mitochondrial membrane potential, reactive oxygen species (ROS) level, ATP content, and the expression of NADPH dehydrogenase subunits 1 (MT-ND1) and NADPH dehydrogenase subunits 6 (MT-ND6) were evaluated. It was observed that compared to the control, the 10 nM MA-5 treatment significantly (p < 0.05) increased total motility (82 ± 3.5% vs. 76 ± 5.9%), progressive motility (67.6 ± 8.2% vs. 51 ± 8.3%), and other parameters (straight-line velocity (VSL), average path velocity (VAP), and curvilinear velocity (VCL)). In addition, 10 nM MA-5 supplementation also improved ram sperm membrane integrity and acrosomal integrity as well increased mitochondrial membrane potential (51.1 ± 0.7% vs. 37.7 ± 1.3%), reduced ROS levels, and elevated adenosine triphosphate (ATP) contents. Furthermore, a Western blot analysis demonstrated that the addition of MA-5 significantly (p < 0.05) increased the expression of MT-ND1 and MT-ND6 proteins in ram sperm, with the 10 nM MA-5 treatment resulting in the highest expression level. These results suggest that MA-5 improves ram sperm quality by maintaining high sperm mitochondrial function during liquid storage at 4 °C.

ALKBH5 is a prognostic factor and promotes the angiogenesis of glioblastoma.

Despite numerous reports indicating the significant impact of RNA modification on malignant glioblastoma (GBM) cell behaviors such as proliferation, invasion and therapy efficacy, its specific involvement in glioblastoma (GBM) angiogenesis is remains unclear and is currently under investigation. In this study, we aimed to investigate the relevance between RNA modification regulators and GBM angiogenesis. Our study employed bioinformatic analyses, including Gene Set Enrichment Analysis (GSEA), differential expression analysis, and Kaplan-Meier survival analysis, to identify regulators of angiogenesis-associated RNA modification (RM). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were applied to identify the enrichment of angiogenesis associated signatures in ALKBH5-high expression GBMs. We also utilized Western blot to verify the upregulation of ALKBH5 in clinical GBM samples. By a series of in vitro and in vivo assays, including plasmid transfection, wound healing, transwell invasion test, tube formation, RT-qPCR, ELISA assays and xenograft mice model, we validated the angiogenesis regulation ability of ALKBH5 in GBM. The N6-methyladenosine (m6A) modification "erase" ALKBH5 emerged as a candidate regulator associated with angiogenesis, demonstrating elevated expression and robust prognostic predictive ability in GBM patients. We also revealed enrichment of vasculature development biological process in GBMs with high ALKBH5 expression. Subsequently, we validated the elevated the expression of ALKBH5 in clinical GBM and paired adjacent tissues through western blot. Additionally, we knocked down the expression of ALKBH5 using sh-RNAs in U87 GBM cells to access the angiogenesis induction ability in U87 cells. In vitro experiments, Human Umbilical Vein Endothelial Cells (HUVECs) were used to perform wound healing, transwell migration and tube formation analysis, results indicated that ALKBH5 knock-down of U87 cells could decrease the pro-angiogenesis ability of U87 GBM cells. Further validation of our bioinformatic findings confirmed that ALKBH5 knockdown impaired VEGFA secretion in both in vitro and in vivo settings in U87 cells. These results comprehensively affirm the crucial role of ALKBH5 in regulating GBM-induced angiogenesis, both in vitro and in vivo. ALKBH5 not only emerges as a promising prognostic factor for GBM patients, but also plays a pivotal role in sustaining GBM progression by promoting angiogenesis.

Pyrroloquinoline Quinone Improves Ram Sperm Quality through Its Antioxidative Ability during Storage at 4 °C.

Sperm motility is an important factor in the migration of sperm from the uterus to the oviduct. During sperm preservation in vitro, sperm generates excessive ROS that damages its function. This study aims to investigate whether the addition of pyrroloquinoline quinone (PQQ) to the diluted medium could improve chilled ram sperm quality, and then elucidates the mechanism. Ram semen was diluted with Tris-citric acid-glucose (TCG) medium containing different doses of PQQ (0 nM, 10 nM, 100 nM, 1000 nM, 10,000 nM), and stored at 4 °C. Sperm motility patterns, plasma membrane integrity, acrosome integrity, mitochondrial membrane potential, reactive oxygen species (ROS) levels, malondialdehyde (MDA) levels, superoxide dismutase (SOD) activity, and ATP levels were measured after preservation. Furthermore, the expressions of NADH dehydrogenase 1 (MT-ND1) and NADH dehydrogenase 6 (MT-ND6) in sperm were also detected by western blotting. In addition, sperm capacitation and the ability of sperm to bind to the zona pellucina were also evaluated. It was observed that the addition of PQQ significantly (p < 0.05) improved ram sperm motility, membrane integrity, and acrosome integrity during preservation. The percentage of sperm with high mitochondrial membrane potential in the PQQ treatment group was much higher than that in the control. In addition, supplementation of PQQ also decreased the sperm MDA and ROS levels, while increasing ATP levels. Interestingly, the levels of MT-ND1 and MT-ND6 protein in sperm treated with PQQ were also higher than that of the control. Furthermore, the addition of 100 nM PQQ to the medium decreased ROS damage in MT-ND1 and MT-ND6 proteins. The addition of 100 nM PQQ significantly (p < 0.05) increased protein tyrosine phosphorylation in ram sperm after induced capacitation. Furthermore, the value of the sperm-zona pellucida binding capacity in the 100 nM PQQ treatment group was also much higher than that of the control. Overall, during chilled ram- sperm preservation, PQQ protected ram sperm quality by quenching the ROS levels to reduce ROS damage and maintain sperm mitochondrial function, and preserved the sperm's high ability of fertilization.

Temperature explains intraspecific functional trait variation in Phragmites australis more effectively than soil properties.

Common reed (Phragmites australis) is a widespread grass species that exhibits a high degree of intraspecific variation for functional traits along environmental gradients. However, the mechanisms underlying intraspecific variation and adaptation strategies in response to environmental gradients on a regional scale remain poorly understood. In this study, we measured leaf, stem, and root traits of common reed in the lakeshore wetlands of the arid and semi-arid regions of the Inner Mongolia Plateau aiming to reveal the regional-scale variation for functional traits in this species, and the corresponding potentially influencing factors. Additionally, we aimed to reveal the ecological adaptation strategies of common reed in different regions using the plant economics spectrum (PES) theory. The results showed that functional-trait variation followed significant latitudinal and longitudinal patterns. Furthermore, we found that these variations are primarily driven by temperature-mediated climatic differences, such as aridity, induced by geographical distance. In contrast, soil properties and the combined effects of climate and soil had relatively minor effects on such properties. In the case of common reed, the PES theory applies to the functional traits at the organ, as well as at the whole-plant level, and different ecological adaptation strategies across arid and semi-arid regions were confirmed. The extent of utilization and assimilation of resources by this species in arid regions was a conservative one, whereas in semi-arid regions, an acquisition strategy prevailed. This study provides new insights into intraspecific variations for functional traits in common reed on a regional scale, the driving factors involved, and the ecological adaptation strategies used by the species. Moreover, it provided a theoretical foundation for wetland biodiversity conservation and ecological restoration.

Surface Enhancement Effects of Tiny SnO2 Nanoparticle Modification on α-Fe2O3 for Room-Temperature NH3 Sensing.

The development of a gas sensor capable of detecting ammonia with high selectivity and rapid response at room temperature has consistently posed a formidable challenge. To address this issue, the present study utilized a one-step solvothermal method to co-assemble α-Fe2O3 and SnO2 by evenly covering SnO2 nanoparticles on the surface of α-Fe2O3. By controlling the morphology and Fe/Sn mole ratio of the composite, the as-prepared sample exhibits high-performance detection of NH3. At room temperature conditions, a gas sensor composed of α-Fe2O3@3%SnO2 demonstrates a rapid response time of 14 s and a notable sensitivity of 83.9% when detecting 100 ppm ammonia. Experiments and density functional theory (DFT) calculations suggest that the adsorption capacity of α-Fe2O3 to ammonia is enhanced by the surface effect provided by SnO2. Meanwhile, the existence of SnO2 tailors the pore structure and effective surface area of α-Fe2O3, creating multiple channels for the diffusion and adsorption of ammonia molecules. Additionally, an N-N heterostructure is formed between α-Fe2O3 and SnO2, which enhances the potential energy barrier and improves the ammonia sensing performance. Demonstration experiments have proved that the sensor shows significant advantages over commercial sensors in the process of ammonia detection in agricultural facilities. This work provides new insights into the perspectives on ammonia detection at room temperature.

Multifunctionality and maintenance mechanism of wetland ecosystems in the littoral zone of the northern semi-arid region lake driven by environmental factors.

The relationship between biodiversity and ecosystem multifunctionality (BEMF) has become an ecological research hot spot in recent years. Changes in biodiversity are non-randomly distributed in space and time in natural ecosystems, and the BEMF relationship is affected by a combination of biotic and abiotic factors. These complex, uncertain relationships are affected by research scale and quantification and measurement indicators. This paper took the Daihai littoral zone wetlands in Inner Mongolia as the research object to reveal the dynamic succession of wetland vegetation and ecosystem function change characteristics and processes during the shrinkage of the lake. The main findings were as follows: the combined effect of aboveground (species and functions) and belowground (bacteria and fungi) diversity was greater than the effect of single components on ecosystem multifunctionality (EMF) (R2 = 80.00 %). Soil salinity (EC) had a direct negative effect on EMF (λ = -0.22), and soil moisture (SM) had a direct positive effect on EMF (λ = 0.19). The results of the hierarchical partitioning analysis showed that plant species richness (Margalef index) was the ideal indicator to explain the EMF and C, N, and P cycling functions in littoral zone wetlands with explanations of 12.25 %, 7.31 %, 7.83 %, and 5.33 %, respectively. The EMF and C and P cycles were mainly affected by bacterial diversity, and the N cycle was mainly affected by fungal abundance in belowground biodiversity. Margalef index and sand content affected EMF through cascading effects of multiple nutrients (FDis, CWMRV, CWMLCC, and bacterial and fungal abundance and diversity) in littoral zone wetlands. This paper provides a reference for exploring the multifunctionality maintenance mechanisms of natural littoral zone wetland ecosystems in the context of global change, and it also provides important theoretical support and basic data for the implementation of ecological restoration in Daihai lake.

Oxidative Radical NHC Catalysis: Divergent Difunctionalization of Olefins through Intermolecular Hydrogen Atom Transfer.

Oxidative N-heterocyclic carbene (NHC) organocatalysis, typically leading to the formation of acyl azolium reactive intermediates, constitutes one of the most important activation strategies for the NHC-catalyzed chemical transformations. Here, we report an unprecedented oxidative radical NHC catalysis by using peroxyester as the external single-electron oxidant to realize divergent difunctionalization of olefins. The key to success of this chemistry is the catalytic generation of oxygen radicals that could trigger an intermolecular hydrogen atom transfer to activate the inert C-H bonds, thereby enabling the productive radical relay process. With this protocol, commonly used general chemicals could serve as radical precursors to allow efficient synthesis of value-added products in a straightforward and cost-effective manner. Preliminary mechanistic investigations, including control experiments and DFT calculations, shed light on the NHC organocatalytic radical reaction mechanism.

Effects of the Cationic Structure on the Adsorption Performance of Ionic Polymers toward Au(III): an Experimental and DFT Study.

Ionic polymers have been proven to be promising adsorbents in recovering Au(III) due to their advantages of simple synthesis and high adsorption efficiency. However, the unclarity of the relationship between the adsorption ability of ionic polymers and their cationic structures hinders further optimization of their adsorption performance. This study synthesized a series of ionic polymers with pyridinium, imidazolium, piperidinium, pyrrolidinium, and triethylammonium cations to discover the effects of the cationic structure on their adsorption properties. Experimental results show that the existence of anion-π interaction between aromatic cations and [AuCl4]- makes the aromatic cations-anion interaction stronger, which does not enhance the adsorption performance of the aromatic-based ionic polymer. This is due to the charge delocalization in the aromatic ring, resulting in a lower electrostatic potential (ESP) of aromatic cations than that of aliphatic cations with a localized charge. The higher the ESP of cations, the better the adsorption performance of the corresponding ionic polymer. This study serves as a deep understanding of the cationic structure-adsorptive performance relationship of the ionic polymer at the molecular level and further provides a theoretical guidance to optimize the adsorption performance of ionic polymers.

Radical Acylalkylation of 1,3-Enynes To Access Allenic Ketones via N-Heterocyclic Carbene Organocatalysis.

An N-heterocyclic carbene organocatalytic 1,4-difunctionalization of 1,3-enynes was developed. This organocatalytic strategy was suitable for a broad spectrum of substrates to efficiently synthesize allenic ketones bearing diverse substituents. Preliminary mechanistic studies suggest a radical reaction pathway for this organocatalytic acylalkylation process.

Controlled fabrication of core-shell γ-Fe2O3@C-Reduced graphene oxide composites with tunable interfacial structure for highly efficient microwave absorption.

The design of a high-performance microwave absorbing material is highly dependent on the synergistic structural design of heterostructure and the appropriate material compositions. Herein, a series of composites of reduced graphene oxide (RGO) and core-shell structured γ-Fe2O3@C nanoparticles have been achieved by a hydrothermal and in-situ chemical vapor deposition (CVD) method. In particular, the structure of the carbon layer, including its graphitization and thickness, can be controlled by optimizing the CVD conditions, which is beneficial to tailor the impedance matching and dielectric loss. The rationally designed RGO/γ-Fe2O3@C composite has multiple electromagnetic dissipation mechanisms. The effective absorption ranges of an optimal sample at a filling rate of 20% can cover 100% X-band and 98% Ku-band at thicknesses of 3.0 mm and 2.2 mm, respectively. This finding suggested that the controllable fabrication of core-shell heterostructures could be viable approach to upgrade the microwave absorption performance of transition metal oxides.

Effect of the length and aromaticity of N3-substituent on adsorption performance of imidazolium-based poly(ionic liquids) towards Pd (II).

Imidazolium-based poly(ionic liquids) (PILs) have been deemed as attractive candidates in the field of precious metal adsorption. However, their further performance optimization is hampered by a lack of an inner understanding of the structure-adsorption performance relationship. In this research, electron and charge distributions of the imidazolium cations are tailored by changing the N3-substitute, and their adsorption performances for PdCl42- were optimized accordingly. Furthermore, the adsorption mechanism is studied by synthesizing corresponding ionic liquid (IL) monomers and their Pd-adducts. Interestingly, longer N3 alkyl chains lead to more hydrogen bonds with PdCl42-, which is beneficial for adsorption. Whereas, it is unfavorable for attracting anions due to a decrease in electrostatic potential (ESP) around cations caused by longer alkyl chains and aromatic substituents at N3 position. It is worth noting that the ESP around the cations plays a more important role in the adsorption process, which determines the adsorption performance of the imidazolium-based PILs. Thus, the performance optimization of imidazolium-based PILs should mainly focus on increasing the ESP of imidazolium cations in the future. This research highlights the potential of the cationic structure-adsorption performance relationship of PILs, which opens a new avenue to develop adsorbents for the metallurgical industry.

[Identifying the Sources of Groudwater NO3--N in Agricultural Region of Qingdao].

To increase crops yields, applying large amounts of fertilizers has become increasingly common in agricultural regions, resulting in NO3--N groundwater pollution. Agricultural non-point pollution is the main source of groundwater NO3--N pollution. To ensure drinking water safety and quality, it is crucial to clarify the sources of NO3--N pollution in agricultural regions. In this study, 35 sampling sites were randomly selected in the Qingdao agricultural area in 2009 and 2019. The spatial distribution of NO3--N concentration was analyzed by the inverse distance weighting method (IDW). The nitrogen and oxygen isotopes were used as a tool to trace sources of NO3--N and the SIAR model was used to quantify contribution proportion of pollution sources. The results showed that the concentration of NO3--N (average) in groundwater in Qingdao has been reduced from 38.49 mg·L-1 in 2009 to 22.37 mg·L-1 in 2019, but it is still higher than the maximum allowable concentration of NO3--N in drinking water set by the World Health Organization (WHO). The NO3--N concentration gradually increased from south to north both in 2009 and 2019. The cross diagram of δ15N-NO3- and δ18O-NO3- show that the main sources of NO3--N in groundwater in Qingdao are chemical fertilizers, soil nitrogen, and manure and sewage. Water isotopes indicate that precipitation was the main source of groundwater in Qingdao. The SIAR model results indicated that the contribution of each source ranked as follows:manure and sewage (47.42%) > soil nitrogen (27.80%) > chemical fertilizer (14.32%) > atmospheric nitrogen depositions (10.43%). From 2009 to 2019, the quality of groundwater in Qingdao has been improved, but NO3--N pollution still cannot be ignored. According to the results, prevention and control should be made to ensure the safety of drinking water and the sustainable development of agriculture.

Properties and mechanism for selective adsorption of Au(iii) on an ionic liquid adsorbent by grafting N-methyl imidazole onto chloromethylated polystyrene beads.

To recover Au(iii) from an acidic chloride-containing solution efficiently, an ionic liquid absorbent (CMPS-IL) was synthesized by grafting N-methyl imidazole onto chloromethylated polystyrene beads (CMPS). The adsorption capacity, selectivity, and reusability were systematically evaluated by a series of adsorption experiments. The maximum adsorption capacity reached up to 516.5 mg g-1 at 318 K. The adsorbent can selectively recover Au(iii) from binary system solutions with a higher separation factor ß Au/M (104-106). Moreover, the adsorption-desorption cycles (7 cycles) showed that the CMPS-IL maintained a stable adsorption performance and high adsorption efficiency. Finally, the adsorption mechanism of CMPS-IL for Au(iii) was investigated by SEM, TEM, XPS, and FT-IR, then proposed with a combination of electrostatic interactions and d-π interaction between imidazolium and AuCl4 -. This study provides an easily-prepared and economical adsorbent for Au(iii) with high selectivity and large adsorption capacity to boost its practical applications.

Magnesium and zinc complexes bearing NNO-tridentate ketiminate ligands: synthesis, structures and catalysis in the ring-opening polymerization of lactides.

A series of magnesium and zinc complexes, [(L1-3)2M2(µ-OBn)2] (M = Mg (1-3), Zn (4-6)), have been synthesized by the reaction of NNO-tridentate ketiminate ligands (L1-3-H) and a stoichiometric amount of MgnBu2 (or ZnEt2) and BnOH. In addition, the reaction of these ketiminate ligands (L1-4-H) with a half equivalent of MgnBu2 (or ZnEt2) in toluene provides [M(L1-4)2] (M = Mg (7-10), Zn (11-14)) in good yields. All of these complexes have been fully characterized by NMR spectroscopy and elemental analysis, and the molecular structures of [(L1)2Mg2(µ-OBn)2] (1), [(L3)2Mg2(µ-OBn)2] (3), [(L1)2Zn2(µ-OBn)2] (4), [Mg(L1)2] (7), [Zn(L1)2] (11) and [Zn(L4)2] (14) have been further confirmed by X-ray crystallographic studies. X-ray diffraction studies revealed that complexes 1, 3 and 4 were dimeric in the solid state, bridging through the benzyloxy groups, while the solid-state structures of 7, 11 and 14 revealed a mononuclear species, six-coordinated by N, N, and O atoms of two ketiminate ligands, forming a distorted octahedron around the metal centre. Complexes 1-6 acted as efficient initiators for the ring-opening polymerization of lactides, producing PLAs with controlled molecular weights and narrow molecular weight distributions. Complex 6 [(L3)2Zn2(µ-OBn)2] exhibited the highest activity towards the ROP of lactides. And complexes 1-6 initiated rac-lactide polymerization to afford heterotactic enriched polymers (Pr up to 0.82). In addition, complexes 7-14 were also shown to efficiently catalyze the ring-opening polymerization of lactides in the presence of BnOH.

Proteome analysis of egg yolk after exposure to zinc oxide nanoparticles.

Zinc oxide nanoparticles (ZnO NPs) are promising candidates as animal diet additive. However, several studies have reported that ZnO NPs cause adverse effects on organisms. Hen egg yolk proteins play vital roles during embryonic development. Although we found ZnO NPs altered the function of the ovary and liver, the effects of ZnO NPs on egg yolk proteins are not as yet understood. In this report, egg yolk proteome was investigated after ZnO NPs treatment. A total of 37 proteins were specifically regulated just by ZnO-NP-50 mg/kg, and 22 proteins were changed solely by ZnSO4-50 mg/kg. Seventeen proteins were regulated by both ZnO-NP-50 mg/kg and ZnSO4-50 mg/kg treatments. Furthermore, the proteins changed by ZnO NPs or ZnSO4 were enriched into different functional groups, respectively, by GO analysis and KEGG pathway enrichment. For the first time, this investigation reports that intact NPs produce a different impact on the egg yolk proteome compared to that of Zn2+. The changes in protein levels by ZnO NPs in egg yolk might influence the value of egg yolk as nutrient and the embryonic development.

The flavonoid TL-2-8 induces cell death and immature mitophagy in breast cancer cells via abrogating the function of the AHA1/Hsp90 complex.

The flavonoid quercetin exhibits significant anticancer activities with few side effects. In the current study, we characterized TL-2-8, a quercetin derivative, as a novel anticancer agent in vitro and in vivo. Cell proliferation and viability were assessed using Cell Counting Kit-8 and CellTiter-Blue assay, respectively. Cell death was examined using PI staining or a TUNEL assay. Mitophagy was determined by measuring autophagic flux and by confocal imaging. Protein expression was examined by Western blotting. We found that TL-2-8 selectively inhibited the proliferation and decreased the viability of various cancer cells (the anti-proliferation IC50 values in MDA-MB-231, MDA-MB-468 and MCF-7 breast cancer cells at 72 h were 8.28, 8.56, and 9.58 µmol/L, respectively), and it displayed only slight cytotoxicity against normal MCF-10A and HEK-293 cells. In MDA-MB-231 and MDA-MB-468 breast cancer cells, TL-2-8 treatment induced the degradation of multiple Hsp90 client proteins without inducing Hsp70. TL-2-8 (3, 6, 12 µmol/L) dose-dependently inhibited the expression of AHA1, an activator of Hsp90 ATPase, and decreased Hsp90-AHA1 complex formation, leading to decreased Hsp90 chaperone function and reduced polo-like kinase 1 (PLK1) signaling. Consequently, impaired mitophagy was induced via the downregulation of lysosomal-associated membrane protein 2 (LAMP2). The in vivo anticancer effects of TL-2-8 were evaluated in an MDA-MB-231 breast cancer xenograft model, which was treated with TL-2-8 (25, 50, 100 mg·kg-1·d-1, po). Administration of TL-2-8 resulted in tumor growth inhibition rates of 37.9%, 58.9% and 70.9%, respectively, whereas quercetin treatment (100 mg·kg-1·d-1, po) produced only a lower tumor growth inhibition rate (49.5%). Furthermore, TL-2-8 treatment significantly extended the lifespan of mice bearing MDA-MB-231 breast cancer cell xenografts. Our results demonstrate that TL-2-8 induces significant cell death and immature mitophagy in breast cancer cells in vitro and in vivo via AHA1 abrogation.

Inhibition of peripubertal sheep mammary gland development by cysteamine through reducing progesterone and growth factor production.

Cysteamine has been used for treating cystinosis for many years, and furthermore it has also been used as a therapeutic agent for different diseases including Huntington's disease, Parkinson's disease (PD), nonalcoholic fatty liver disease, malaria, cancer, and others. Although cysteamine has many potential applications, its use may also be problematic. The effects of low doses of cysteamine on the reproductive system, especially the mammary glands are currently unknown. In the current investigation, low dose (10 mg/kg BW/day) of cysteamine did not affect sheep body weight gain or organ index of the liver, spleen, or heart; it did, however, increase the levels of blood lymphocytes, monocytes, and platelets. Most interestingly, it inhibited mammary gland development after 2 or 5 months of treatment by reducing the organ index and the number of mammary gland ducts. Plasma growth hormone and estradiol remained unchanged; however, plasma progesterone levels and the protein level of HSD3ß1 in sheep ovaries were decreased by cysteamine. In addition to steroid hormones, growth factors produced in the mammary glands also play crucial roles in mammary gland development. Results showed that protein levels of HGF, GHR, and IGF1R were decreased after 5 months of cysteamine treatment. These findings together suggest that progesterone and local growth factors in mammary glands might be involved in cysteamine initiated inhibition of pubertal ovine mammary gland development. Furthermore, it may lead to a reduction in fertility. Therefore, cysteamine should be used with great caution until its actions have been further investigated and its limitations overcome.

Regulation of steroid hormones and energy status with cysteamine and its effect on spermatogenesis.

Although it is well known that cysteamine is a potent chemical for treating many diseases including cystinosis and it has many adverse effects, the effect of cysteamine on spermatogenesis is as yet unknown. Therefore the objective of this investigation was to explore the effects of cysteamine on spermatogenesis and the underlying mechanisms. Sheep were treated with vehicle control, 10mg/kg or 20mg/kg cysteamine for six months. After that, the semen samples were collected to determine the spermatozoa motility by computer-assisted sperm assay method. Blood samples were collected to detect the levels of hormones and the activity of enzymes. Spermatozoa and testis samples were collected to study the mechanism of cysteamine's actions. It was found that the effects of cysteamine on spermatogenesis were dose dependent. A low dose (10mg/kg) cysteamine treatment increased ovine spermatozoa motility; however, a higher dose (20mg/kg) decreased both spermatozoa concentration and motility. This decrease might be due to a reduction in steroid hormone production by the testis, a reduction in energy in the testis and spermatozoa, a disruption in the blood-testis barrier, or a breakdown in the vital signaling pathways involved in spermatogenesis. The inhibitory effects of cysteamine on sheep spermatogenesis may be used to model its effects on young male patients with cystinosis or other diseases that are treated with this drug. Further studies on spermatogenesis that focus on patients treated with cysteamine during the peripubertal stage are warranted.

Bilobetin ameliorates insulin resistance by PKA-mediated phosphorylation of PPARα in rats fed a high-fat diet.

BACKGROUND AND PURPOSE: The amelioration of insulin resistance by bilobetin is closely related to its hypolipidaemic effect. The aim of the present study was to determine the insulin-sensitizing mechanism of bilobetin by elucidating its effect on lipid metabolism. EXPERIMENTAL APPROACH: Rats fed a high-fat diet were treated with bilobetin for either 4 or 14 days before applying a hyperinsulinaemic-euglycaemic clamp. Triglyceride and fatty acids labelled with radioactive isotopes were used to track the transportation and the fate of lipids in tissues. The activity of lipid metabolism-related enzymes and ß-oxidation rate were measured. Western blot was used to investigate the phosphorylation, translocation and expression of PPARα in several tissues and cultured cells. The location of amino acid residues subjected to phosphorylation in PPARα was also studied. KEY RESULTS: Bilobetin ameliorated insulin resistance, increased the hepatic uptake and oxidation of lipids, reduced very-low-density lipoprotein triglyceride secretion and blood triglyceride levels, enhanced the expression and activity of enzymes involved in ß-oxidation and attenuated the accumulation of triglycerides and their metabolites in tissues. Bilobetin also increased the phosphorylation, nuclear translocation and activity of PPARα accompanied by elevated cAMP level and PKA activity. Threonine-129-alanine and/or serine-163-alanine mutations on the PPARα genes and PKA inhibitors prevented the effects of bilobetin on PPARα. However, cells overexpressing PKA appeared to stimulate the phosphorylation, nuclear translocation and activity of PPARα. CONCLUSIONS AND IMPLICATIONS: Bilobetin treatment ameliorates hyperlipidaemia, lipotoxicity and insulin resistance in rats by stimulating PPARα-mediated lipid catabolism. PKA activation is crucial for this process.

Altering intercomponent interactions in a photochromic multi-state [2]rotaxane.

Rotaxanes have attracted much attention because of their challenging constructions and potential applications. In this paper, a multi-state [2]rotaxane, in which a dithienylethene-functionalized dibenzo-24-crown-8 macrocycle was interlocked onto a thread component bearing a 4-morpholin-naphthalimide fluorescent stopper and two distinct recognition sites, namely, dibenzylammonium and N-methyltriazolium recognition sites, was prepared and studied. By introducing a dithienylethene photochrome into the macrocycle component, multi-mode alteration of the intercomponent interactions, such as energy transfer, electron transfer, and charge transfer interaction between the photochrome and the fluorescent naphthalimide stopper could be altered in this multi-state rotaxane system in response to the combination of chemical and photochemical stimuli.