Hypoxia Aggravates Neuron Ferroptosis in Early Brain Injury Following Subarachnoid Hemorrhage via NCOA4-Meditated Ferritinophagy.

The occurrence of early brain injury (EBI) significantly contributes to the unfavorable prognosis observed in patients with subarachnoid hemorrhage (SAH). During the process of EBI, a substantial quantity of iron permeates into the subarachnoid space and brain tissue, thereby raising concerns regarding its metabolism. To investigate the role and metabolic processes of excessive iron in neurons, we established both in vivo and in vitro models of SAH. We substantiated that ferritinophagy participates in iron metabolism disorders and promotes neuronal ferroptosis using an in vivo model, as detected by key proteins such as ferritin heavy chain 1, glutathione peroxidase 4, autophagy related 5, nuclear receptor coactivator 4 (NCOA4), LC3B, and electron microscopy results. By interfering with NCOA4 expression in vitro and in vivo, we confirmed the pivotal role of elevated NCOA4 levels in ferritinophagy during EBI. Additionally, our in vitro experiments demonstrated that the addition of oxyhemoglobin alone did not result in a significant upregulation of NCOA4 expression. However, simultaneous addition of oxyhemoglobin and hypoxia exposure provoked a marked increase in NCOA4 expression and heightened ferritinophagy in HT22 cells. Using YC-1 to inhibit hypoxia signaling in in vitro and in vitro models effectively attenuated neuronal ferroptosis. Collectively, we found that the hypoxic microenvironment during the process of EBI exaggerates iron metabolism abnormalities, leading to poor prognoses in SAH. The findings also offer a novel and potentially effective foundation for the treatment of SAH, with the aim of alleviating hypoxia.

Pairing with Enriched Sound Exposure Restores Auditory Processing Degraded by an Antidepressant.

Antidepressants, while effective in treating depression and anxiety disorders, also induce deficits in sensory (particularly auditory) processing, which in turn may exacerbate psychiatric symptoms. How antidepressants cause auditory signature deficits remains largely unknown. Here, we found that fluoxetine-treated adult female rats were significantly less accurate when performing a tone-frequency discrimination task compared with age-matched control rats. Their cortical neurons also responded less selectively to sound frequencies. The degraded behavioral and cortical processing was accompanied by decreased cortical perineuronal nets, particularly those wrapped around parvalbumin-expressing inhibitory interneurons. Furthermore, fluoxetine induced critical period-like plasticity in their already mature auditory cortices; therefore, a brief rearing of these drug-treated rats under an enriched acoustic environment renormalized auditory processing degraded by fluoxetine. The altered cortical expression of perineuronal nets was also reversed as a result of enriched sound exposure. These findings suggest that the adverse effects of antidepressants on auditory processing, possibly because of a reduction in intracortical inhibition, can be substantially alleviated by simply pairing drug treatment with passive, enriched sound exposure. They have important implications for understanding the neurobiological basis of antidepressant effects on hearing and for designing novel pharmacological treatment strategies for psychiatric disorders.SIGNIFICANCE STATEMENT Clinical experience suggests that antidepressants adversely affect sensory (particularly auditory) processing, which can exacerbate patients' psychiatric symptoms. Here, we show that the antidepressant fluoxetine reduces cortical inhibition in adult rats, leading to degraded behavioral and cortical spectral processing of sound. Importantly, fluoxetine induces a critical period-like state of plasticity in the mature cortex; therefore, a brief rearing under an enriched acoustic environment is sufficient to reverse the changes in auditory processing caused by the administration of fluoxetine. These results provide a putative neurobiological basis for the effects of antidepressants on hearing and indicate that antidepressant treatment combined with enriched sensory experiences could optimize clinical outcomes.

Coeffect of pyrolysis temperature and potassium phosphate impregnation on characteristics, stability, and adsorption mechanism of phosphorus-enriched biochar.

Potassium phosphate (K3PO4)-impregnated bamboo was pyrolyzed at temperatures ranging from 350 to 950 °C to explore the coeffect of pyrolysis temperature and K3PO4 impregnation on biochar's characteristics and adsorption behavior. The degree of aromatization and graphitization in phosphorus-enriched biochars (PRBCs) rose as temperature increased, whereas H/C and O/C ratios, pH value, and O-containing group content decreased. The pre-aging impact of K3PO4 impregnation results in increased stability and adsorption performance of PRBCs. Adsorption mechanism of PRBCs to heavy metal varies from pyrolysis temperature. Micropores dominate medium-temperature PRBCs (prepared at 550 âˆ¼ 750 °C), possessing the highest P-containing group content (116 % that of PRBC-350) and maximal adsorption capacity (greater than289 mg/g). The medium-temperature PRBCs adsorb Cd (II) via the role of O-containing groups, PO43-, and P2O74-, mainly by reactions of organic complexation, precipitation and inorganic complexation, respectively. 550 °C is the optimal pyrolysis temperature for both energy saving and heavy metal adsorption.

Application of 1H-MRS in end-stage renal disease with depression.

BACKGROUND: To investigate the metabolite changes in the frontal lobe of the end-stage renal disease (ESRD) patients with depression using proton magnetic resonance spectroscopy (1H-MRS). METHODS: All subjects were divided into three groups: ESRD patients with depression (30 cases), ESRD patients without depression (27 cases) and 32 normal subjects. ESRD with depression patients were further divided into two groups according to the severity of depression: 14 cases of ESRD with severe depression group (Hamilton Depression Rating Scale (HAMD) score ≥ 35) and 16 cases of ESRD with mild to moderate depression group (20 ≤ HAMD score<35). 1H-MRS was used in brain regions of all subjects to measure N-acetylaspartate/creatine (NAA/Cr), choline-containing compounds/creatine (Cho/Cr) and myo-inositol/creatine (MI/Cr) ratios of the frontal lobe. Correlations between the metabolite ratio and HAMD score as well as clinical finding were confirmed, respectively. RESULTS: ESRD patients with depression showed lower NAA/Cr ratio and higher Cho/Cr ratio compared with ESRD patients without depression and normal subjects. NAA/Cr ratio was negatively correlated with the HAMD score. Cho/Cr ratio was positively correlated with the HAMD score. There were positive correlations between NAA/Cr ratio and blood urea notrogen (BUN) as well as creatinine (CRE) concentration, respectively. There was a negative correlation between Cho/Cr ratio and sodium concentration. The Cho/Cr ratio was positively correlated with the potassium concentration. CONCLUSIONS: MR spectroscopy identified some metabolite changes in ESRD patients with depression.

Synthesis, characterization of phosphorus-containing copolyester and its application as flame retardants for poly(butylene succinate) (PBS).

Two novel phosphorus-containing copolyesters were synthesized by direct polycondensation reaction of phenyl dichlorophosphate, 1,4-succinic acid and 1,4-butanediol using stannous chloride (SnCl2) and 4-Methylbenzenesulfonic acid as catalyst, and its chemical structures were identified by 1H and 31P nuclear magnetic resonances (1H and 31P NMR). The resulting phosphorus-containing poly(butylene succinate) (PPBS) was characterized by X-ray diffraction (XRD), polarized optical microscope (POM), thermogravimetry analysis (TGA) and differential scanning calorimetry (DSC). PPBS can be as a flame retardant for commercial poly(butylene succinate) (PBS). A series of flame retardant composite materials were produced by melt-blending of PBS and PPBS. The comprehensive flame retardant property of composite materials was evaluated by limited oxygen index (LOI). While 20 wt % of PPBS was added into PBS resin, good flame retardant properties could be obtained. Composite materials can have much better flame retardancy (LOI = 30) than neat PBS resin. Thermogravimetric analysis (TGA) showed that the weight loss of PBS was decreased by the introduction of PPBS. In addition, possible flame retardancy mechanism of PPBS in composite materials was analyzed by SEM photos of char residue.

High fish oil diet promotes liver inflammation and activates the complement system.

Diets rich in n-3 polyunsaturated fatty acid (n-3 PUFA) fish oil (FO) have beneficial effects in obesity­associated metabolic disease. However, contradictory roles in inflammatory disease intervention have been reported. Our previous work revealed that a high­FO diet promoted myeloid cell differentiation by modifying the bone marrow microenvironment; however, its effects on liver inflammation and complement system activation remain unknown. By performing ELISA, reverse transcription­quantitative polymerase chain reaction, flow cytometry and histology on mice fed with high­FO and low­fat diets, the present study demonstrated that a 4­week high­FO diet promoted liver inflammation in mice without affecting body or liver weight. The livers of high­FO diet mice exhibited increased infiltration of T cells and CD11b+ Gr­1+ myeloid cells. Additionally, a higher level of IL­1ß and MCP­1 mRNA expression was detected, suggesting that the high­FO diet promoted liver inflammation. Further experiments indicated that the high­FO diet increased the total hemolytic complement activity (CH50), promoted the production of the membrane attack complex and increased the levels of various complement proteins in vivo, including complement components C3, C4b, C1qb and factor B. Furthermore, higher concentrations of triglyceride were detected in the peripheral blood of high­FO diet mice, indicating the potential protective roles of n­3 PUFAs in FO against lipotoxicity in hyperlipidemia. Collectively, the present study demonstrated that high FO intake induced inflammation and activated the complement system in the liver. However, further study is required to determine the exact mechanisms.

Apoptosis of rat hepatic stellate cells induced by diallyl trisulfide and proteomics profiling in vitro.

Diallyl trisulfide (DATS), a major garlic derivative, inhibits cell proliferation and triggers apoptosis in a variety of cancer cell lines. However, the effects of DATS on hepatic stellate cells (HSCs) remain unknown. The aim of this study was to analyze the effects of DATS on cell proliferation and apoptosis, as well as the protein expression profile in rat HSCs. Rat HSCs were treated with or without 12 and 24 µg/mL DATS for various time intervals. Cell proliferation and apoptosis were determined using tetrazolium dye (MTT) colorimetric assay, bromodeoxyuridine (5-bromo-2'-deoxyuridine; BrdU) assay, Hoechst 33342 staining, electroscopy, and flow cytometry. Protein expression patterns in HSCs were systematically studied using 2-dimensional electrophoresis and mass spectrometry. DATS inhibited cell proliferation and induced apoptosis of HSCs in a time-dependent manner. We observed clear morphological changes in apoptotic HSCs and dramatically increased annexin V-positive - propidium iodide negative apoptosis compared with the untreated control group. Twenty-one significant differentially expressed proteins, including 9 downregulated proteins and 12 upregulated proteins, were identified after DATS administration, and most of them were involved in apoptosis. Our results suggest that DATS is an inducer of apoptosis in HSCs, and several key proteins may be involved in the molecular mechanism of apoptosis induced by DATS.

Effect of Rhizoma Polygoni Cuspidati and Ramulus Cinnamomi compatibility on uric acid metabolism and urinary neutrophil gelatinase-associated lipocalin and kidney injury molecule-1 in rats with hyperuricemia.

OBJECTIVE: To explore the effects of Rhizoma Polygoni Cuspidati and Ramulus Cinnamomi compatibility (PR) on uric acid metabolism and the expression of urinary neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) in rats with hyperuricemia. METHODS: Seventy male Sprague Dawley (SD) rats were randomly divided into 7 groups with 10 rats per group, including the normal group, model group, allopurinol group, benzbromarone group and PR groups at 3 doses (3.5, 7, 14 g/kg). Except the normal group, rats of the other groups were intragastrically administered 100 mg/kg hypoxanthine and 250 mg/kg ethambutol, and subcutaneously injected with 200 mg/kg potassium oxonate. All rats were continuously modeled for 17 days, and gavaged with corresponding drugs. The rats of the normal and model groups were gavaged with saline, once a day, for 2 weeks. The levels of serum uric acid (SUA), blood urea nitrogen (BUN) and creatinine (Cr) were determined. In addition, the contents of NGAL and KIM-1 in urine and the mRNA and protein expressions of xanthine oxidase (XOD) in liver of hyperuricemia rats were measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot, respectively. Moreover, the pathological changes of kidney were analyzed by hematoxylin and eosin (HE) stain method. RESULTS: Compared with the normal group, the levels of SUA, BUN, NGAL and KIM-1 and the expressions of hepatic XOD mRNA and protein in the hyperuricemia rats were increased signifificantly (P<0.01). PR signifificantly decreased the levels of SUA, BUN, NGAL and KIM-1 and down-regulated the mRNA and protein expressions of hepatic XOD (P<0.05 or P<0.01). In addition, the pathological changes of kidney were signifificantly suppressed by oral administration of PR. CONCLUSIONS: PR ameliorated uric acid metabolism and protected renal function, the underlying mechanism was mediated by decreasing the levels of SUA, BUN, NGAL and KIM-1, inhibiting the expression of hepatic XOD and ameliorating the pathological change of kidney.

Deactivation of Association Cortices Disrupted the Congruence of Visual and Auditory Receptive Fields in Superior Colliculus Neurons.

Physiological and behavioral studies in cats show that corticotectal inputs play a critical role in the information-processing capabilities of neurons in the deeper layers of the superior colliculus (SC). Among them, the sensory inputs from functionally related associational cortices are especially critical for SC multisensory integration. However, the underlying mechanism supporting this influence is still unclear. Here, results demonstrate that deactivation of relevant cortices can both dislocate SC visual and auditory spatial receptive fields (RFs) and decrease their overall size, resulting in reduced alignment. Further analysis demonstrated that this RF separation is significantly correlated with the decrement of neurons' multisensory enhancement and is most pronounced in low stimulus intensity conditions. In addition, cortical deactivation could influence the degree of stimulus effectiveness, thereby illustrating the means by which higher order cortices may modify the multisensory activity of SC.

Structure-activity relationship studies and discovery of a potent transient receptor potential vanilloid (TRPV1) antagonist 4-[3-chloro-5-[(1S)-1,2-dihydroxyethyl]-2-pyridyl]-N-[5-(trifluoromethyl)-2-pyridyl]-3,6-dihydro-2H-pyridine-1-carboxamide (V116517) as a clinical candidate for pain management.

A series of novel tetrahydropyridinecarboxamide TRPV1 antagonists were prepared and evaluated in an effort to optimize properties of previously described lead compounds from piperazinecarboxamide series. The compounds were evaluated for their ability to block capsaicin and acid-induced calcium influx in CHO cells expressing human TRPV1. The most potent of these TRPV1 antagonists were further characterized in pharmacokinetic, efficacy, and body temperature studies. On the basis of its pharmacokinetic, in vivo efficacy, safety, and toxicological properties, compound 37 was selected for further evaluation in human clinical trials.

Astaxanthin offers neuroprotection and reduces neuroinflammation in experimental subarachnoid hemorrhage.

BACKGROUND: Neuroinflammation has been proven to play a crucial role in early brain injury pathogenesis and represents a target for treatment of subarachnoid hemorrhage (SAH). Astaxanthin (ATX), a dietary carotenoid, has been shown to have powerful anti-inflammation property in various models of tissue injury. However, the potential effects of ATX on neuroinflammation in SAH remain uninvestigated. The goal of this study was to investigate the protective effects of ATX on neuroinflammation in a rat prechiasmatic cistern SAH model. METHODS: Rats were randomly distributed into multiple groups undergoing the sham surgery or SAH procedures, and ATX (25 mg/kg or 75 mg/kg) or equal volume of vehicle was given by oral gavage at 30 min after SAH. All rats were sacrificed at 24 h after SAH. Neurologic scores, brain water content, blood-brain barrier permeability, and neuronal cell death were examined. Brain inflammation was evaluated by means of expression changes in myeloperoxidase, cytokines (interleukin-1ß, tumor necrosis factor-α), adhesion molecules (intercellular adhesion molecule-1), and nuclear factor kappa B DNA-binding activity. RESULTS: Our data indicated that post-SAH treatment with high dose of ATX could significantly downregulate the increased nuclear factor kappa B activity and the expression of inflammatory cytokines and intercellular adhesion molecule-1 in both messenger RNA transcription and protein synthesis. Moreover, these beneficial effects lead to the amelioration of the secondary brain injury cascades including cerebral edema, blood-brain barrier disruption, neurological dysfunction, and neuronal degeneration. CONCLUSIONS: These results indicate that ATX treatment is neuroprotective against SAH, possibly through suppression of cerebral inflammation.

Resveratrol prevents neuronal apoptosis in an early brain injury model.

BACKGROUND: Resveratrol has been shown to attenuate cerebral vasospasm after subarachnoid hemorrhage (SAH); however, no study has explored its neuroprotective effect in early brain injury (EBI) after experimental SAH. The aim of this study was to evaluate the antiapoptotic function of resveratrol in EBI and its relationship with the PI3K/Akt survival pathway. METHODS: Experimental SAH was induced in adult male rats by prechiasmatic cistern injection. Control and SAH rats were divided into six groups and treated with low (20 mg/kg) or high (60 mg/kg) concentrations of resveratrol with or without LY294002 cotreatment. Brain samples of the rats were analyzed by immunohistochemistry, immunofluorescence staining, Western blotting, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) apoptosis assays. RESULTS: High-concentration but not low-concentration resveratrol treatment in SAH rats led to a significant increase in phosphorylated Akt (p-Akt) protein levels compared with SAH rats without treatment. In addition, p-Akt-positive cells mainly colocalized with NeuN-positive cells. Neuronal apoptosis in SAH rat brain was attenuated by high-concentration resveratrol treatment. The antiapoptotic effect of resveratrol in SAH rats could be partially abrogated by the PI3K/Akt signaling inhibitor LY294002. CONCLUSIONS: Our results show that resveratrol has an antiapoptotic effect in EBI and that resveratrol might act through the PI3K/Akt signaling pathway.

Refining cortical representation of sound azimuths by auditory discrimination training.

Although training-based auditory cortical plasticity in the adult brain has been previously demonstrated in multiparametric sound domains, neurochemical mechanisms responsible for this form of plasticity are not well understood. In this study, we trained adult rats to identify a target sound stimulus at a specific azimuth angle by using a reward-contingent auditory discrimination task. We found that auditory spatial discrimination training significantly enhanced representation of sound azimuths in the primary auditory cortex, as shown by sharper azimuth-selective curves and more evenly distributed best angles of cortical neurons. Training also facilitated long-term potentiation of field potentials in the primary auditory cortex induced by theta burst stimulation of the white matter. In parallel, there were significant alterations in expression levels of certain cortical GABA(A) and NMDA receptor subunits, resulting in a marked decrease in the level of GABA(A) relative to NMDA receptors. These changes in the expression profile of inhibitory and excitatory neurotransmitter receptor subunits might enhance synaptic transmission, thereby facilitating training-induced cortical plasticity in the spatial domain.

Natural restoration of critical period plasticity in the juvenile and adult primary auditory cortex.

Since its first description >40 years ago, the neurological "critical period" has been predominantly described as an early, plastic postnatal brain development stage that rather abruptly advances to an aplastic or less plastic "adult" stage. Here, we show that chronic exposure of juvenile or adult rats to moderate-level acoustic noise results in a broad reversal of maturational changes that mark the infant-to-adult progression in the primary auditory cortex. In time, noise exposure reinstates critical period plasticity. Cortical changes resulting from noise exposure are again reversed to reestablish a physically and functionally normal adult cortex, by returning animals to natural acoustic environments. These studies show that at least some of neurological changes believed to mark the transition from the infantile to the mature (adult) stage are, by their nature, reversible.

Successive-signal biasing for a learned sound sequence.

Adult rats were trained to detect the occurrence of a two-element sound sequence in a background of nine other nontarget sound pairs. Training resulted in a modest, enduring, static expansion of the cortical areas of representation of both target stimulus sounds. More importantly, once the initial stimulus A in the target A-B sequence was presented, the cortical "map" changed dynamically, specifically to exaggerate further the representation of the "anticipated" stimulus B. If B occurred, it was represented over a larger cortical area by more strongly excited, more coordinated, and more selectively responding neurons. This biasing peaked at the expected time of B onset with respect to A onset. No dynamic biasing of responses was recorded for any sound presented in a nontarget pair. Responses to nontarget frequencies flanking the representation of B were reduced in area and in response strength only after the presentation of A at the expected time of B onset. This study shows that cortical areas are not representationally static but, to the contrary, can be biased moment by moment in time as a function of behavioral context.

Rapid and sensitive detection of point mutation by DNA ligase-based electrochemiluminescence assay.

The identification of single-base mutations in particular genes plays an increasingly important role in medical diagnosis and prognosis of genetic-based diseases. Here we report a new method for the analysis of point mutations in genomic DNA through the integration of allele-specific oligonucleotide ligation assay (OLA) with magnetic beads-based electrochemiluminescence (ECL) detection scheme. In this assay, the tris(bipyridine) ruthenium (TBR) labeled probe and the biotinylated probe are designed to perfectly complementary to the mutant target; thus a ligation can be generated between those two probes by Taq DNA Ligase in the presence of mutant target. If there is an allele mismatch, the ligation does not take place. The ligation products are then captured onto streptavidin-coated paramagnetic beads, and detected by measuring the ECL signal of the TBR label. Results showed that the new method held a low detection limit down to 10 fmol and was successfully applied in the identification of point mutations from ASTC-alpha-1 cell line, PANC-1 cell line and blood cell in codon 273 of TP53 oncogene. In summary, this method provides a sensitive, cost-effective and easy operation approach for point mutation detection.

[A case-control study on drinking green tea and decreasing risk of cancers in the alimentary canal among cigarette smokers and alcohol drinkers].

OBJECTIVE: To explore the role of green tea in decreasing the risks of gastric cancer, liver cancer, esophageal cancer among alcohol drinkers or cigarette smokers. METHODS: A population based case-control study was conducted in Taixing, Jiangsu province. RESULTS: In Taixing city, identified cases of stomach, liver and esophageal cancers were chosen with informed consent. The numbers were 206, 204, 218 respectively. Controls were chosen from normal population having lived in the area for longer than 10 years, also with informed consent. Green tea drinking seemed to have decreased 81%, 78%, 39% risk for the development of gastric cancer, liver cancer and esophageal cancer among alcohol drinkers. It might also have decreased 16%, 43%, 31% on the risks of developing the three kinds of cancers among cigarette smokers. Interaction assessment showed that drinking green tea could significantly decrease the risk of gastric cancer and liver cancer among alcohol drinkers, with ORs of interaction item 0.23 (95% CI: 0.10 - 0.55) and 0.25 (95% CI: 0.11 - 0.57) respectively. CONCLUSION: Habit of drinking green tea seemed to have significant protective effects on the development of both gastric and liver cancer among alcohol drinkers while, green tea also having some protective effect on esophageal cancer among alcohol drinkers and on three kinds of cancers among cigarette smokers.