MET Inhibits the Proliferation of EC Cells by Increasing MPA Sensitivity.

Context: The high resistance rate and high recurrence rate of progesterone only as a treatment for endometrial cancer (EC) limit its clinical application. Metformin (MET) may have antitumor ability. Combining MET and medroxyprogesterone acetate (MPA) may strengthen their inhibitory effects on proliferation of EC cells, but MET's mechanisms remain unclear. Objective: The study intended to identify the specific molecular mechanism that MET combined with MPA uses against EC progression. Design: The research team performed a controlled animal study. Setting: The study took place at Xuzhou Medical University in Xuzhou, China. Animals: The animals were16 female non-obese diabetic-severe combined immunodeficient (NOD-SCID) nude mice, about 12 to 16 g in weight. Interventions: The research team divided randomly, the mice into four groups and induced EC in all groups, four in each group: (1) The control group which received received normal saline, (2) the MPA group, which received 100 mg/kg of MPA; (3) the MET group, which received metformin at the rate of 200 mg/kg, each gavage volume was 0.1ml; (4) the MET+MPA group, which received 100 mg/kg of MPA and 200 mg/kg of MET. Outcome measures: The research team: (1) used a CCK-8 kit, an EdU assay, and a flow-cytometry assay to measure cancer-cell proliferation, count, and viability; determine the cell cycle; and measure apoptosis; (2) performed a Western blot analysis to determine the expression of the PR, CD133, pAkt, totalAkt, p-mTOR, and totalTOR antibodies; and (3) determined the size and volume of tumors in vivo and used immunohistochemical staining to determine expression of the Ki67 protein. Results: The MET+MPA group had a significantly lower number of cancer cells than the MET or MDA groups (both P < .001). That group also had significantly more stagnated cancer cells in the G0/G1 phase and significantly fewer cancer cells in the S phase or G2/M phase control, MET, or MPA groups (all P < .01). The MET+MPA group's PCNA and Ki-67 protein expression was significantly lower than that of the MET and MPA group. The EDU assay yielded similar results. Additionally, the MET+MPA group had significantly higher PR expression than that of to MET or MPA group (both P < .001). The MET and MPA groups' expression of CD133, p-Akt, and p-mTOR were significantly lower than those of the control group, while the MET+MPA group's levels were significantly lower than those of the MET and MPA groups. In-vivo experiments revealed that the MET and MPA groups did show decreased tumor size and volume. The MET+MPA group had tumor weights that were significantly lower and tumor volumes were significantly smaller than those of the MET and MPA groups (all P < .001). Immunohistochemical analysis revealed that the MET+MPA group's levels of the Ki-67 antigen were significantly lower than those of the MET and MPA groups. Conclusions: MET inhibited the proliferation of EC cells by increasing MPA-sensitivity, which was dependent on the inhibition of the CD133 expression and the Akt/mTOR pathway. In addition, if MET acts as an effective progestin sensitizer, it certainly offers promising therapeutic prospects for patients with early-stage EC or overgrown endometrium who have fertility requirements.

Mucilaginibacter gilvus sp. nov., isolated from Antarctic soil.

A Gram-stain-negative, aerobic, non-motile, rod-shaped, cold-tolerant bacterium, designated F01003T, was isolated from soil sampled near Happiness Bay on the west coast of Antarctica. Strain F01003T was found to grow at 4-30 °C (optimum, 25 °C), pH 5.5-8.0 (pH 6.5-7.0) and in the presence of 0-1 % NaCl (0 %, w/v). Cells were oxidase-positive and catalase-positive. Strain F01003T contained menaquinone 7 (MK-7) as the predominant respiratory quinone. The main cellular fatty acids included summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c) and iso-C15 : 0. Phosphatidylethanolamine and an unidentified aminolipid were identified as the major polar lipids. The DNA G+C content of strain F01003T was 44.8 mol%. Phylogenetic analysis of the nearly full-length 16S rRNA gene sequence revealed that strain F01003T was most closely related to the genus Mucilaginibacter and exhibited the highest sequence similarity to Mucilaginibacter phyllosphaerae LMG 29118T (97.3 %). On the basis of the evidence presented in this polyphasic taxonomic study, strain F01003T is considered to represent a novel species of the genus Mucilaginibacter, for which the name Mucilaginibactergilvus sp. nov. is proposed. The type strain is F01003T (=KCTC 62991T=CCTCC AB 2019023T).

Ultrasound-to-plastic optical fiber coupling as an acoustic source probe.

An integrated ultrasonic detection system that consists of emission and detection has been proposed and demonstrated experimentally. The proposed emission source is based on plastic optical fiber (POF). In order to promote the coupling of ultrasound-to-POF, and the end of the POF is heated to form a circular pedestal. A homemade fiber Fabry-Perot interferometer is employed to evaluate the coupling efficiency of ultrasound-to-POF. The sensing head is spliced with a short section of hollow-core fiber and single-mode fiber, resulting in an air microbubble formation by discharging. The experimental results show that ultrasound can be transmitted effectively in a narrow space using the coupling method, and the compact sensor also presents a considered sensitivity for ultrasonic detection. This all-fiber ultrasonic interrogation can be integrated as a system for the application in the bioimaging field, especially the organism body.

Stress and corticosterone alter synaptic plasticity in a rat model of Parkinson's disease.

As a major influence on neuronal function and plasticity, chronic stress can affect the progression and symptoms of neurodegenerative conditions, such as Parkinson's disease (PD). Here we investigated the influence of unilateral dopamine depletion and stress on dopamine-related hallmarks of stress response and neuronal plasticity in a rat model of PD. Animals received either restraint stress or a combination of adrenalectomy and corticosterone (CORT) supplementation to clamp circulating glucocorticoid levels for three weeks prior to unilateral nigrostriatal dopamine depletion. Rats were tested in skilled and non-skilled motor function up to three weeks post-lesion. Midbrain mRNA expression assessments included markers of dopamine function and neuroplasticity, such as tyrosine hydroxylase (TH), synaptophysin (SYN), calcyon, and glucocorticoid receptor (GR). Along with impaired motor performance, stress and clamped CORT partially preserved TH expression in both substantia nigra (SN) and ventral tegmental area (VTA), but differentially modulated the expression of SYN, calcyon, and GR mRNA in midbrain and cortical areas. Stress reduced synaptophysin mRNA expression in SN/VTA, and elevated calcyon mRNA optical density in both non-lesion and lesion hemispheres. Stress and CORT increased GR mRNA in the non-lesion SN/VTA, while in the lesion hemisphere GR mRNA was only elevated by CORT. In the motor cortex and striatum, however, GR was higher in both hemispheres under both experimental conditions. These findings suggest that stress and stress hormones differentially affect dopaminergic function and neuroplasticity in a rat model of PD. The findings suggest a role for stress in motor and non-motor symptoms of PD and stress response.

UW Imaging of Seismic-Physical-Models in Air Using Fiber-Optic Fabry-Perot Interferometer.

A fiber-optic Fabry-Perot interferometer (FPI) has been proposed and demonstrated for the ultrasound wave (UW) imaging of seismic-physical models. The sensor probe comprises a single mode fiber (SMF) that is inserted into a ceramic tube terminated by an ultra-thin gold film. The probe performs with an excellent UW sensitivity thanks to the nanolayer gold film, and thus is capable of detecting a weak UW in air medium. Furthermore, the compact sensor is a symmetrical structure so that it presents a good directionality in the UW detection. The spectral band-side filter technique is used for UW interrogation. After scanning the models using the sensing probe in air, the two-dimensional (2D) images of four physical models are reconstructed.

Environmental Intervention as a Therapy for Adverse Programming by Ancestral Stress.

Ancestral stress can program stress sensitivity and health trajectories across multiple generations. While ancestral stress is uncontrollable to the filial generations, it is critical to identify therapies that overcome transgenerational programming. Here we report that prenatal stress in rats generates a transgenerationally heritable endocrine and epigenetic footprint and elevated stress sensitivity which can be alleviated by beneficial experiences in later life. Ancestral stress led to downregulated glucocorticoid receptor and prefrontal cortex neuronal densities along with precocious development of anxiety-like behaviours. Environmental enrichment (EE) during adolescence mitigated endocrine and neuronal markers of stress and improved miR-182 expression linked to brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) regulation in stressed lineages. Thus, EE may serve as a powerful intervention for adverse transgenerational programming through microRNA-mediated regulation of BDNF and NT-3 pathways. The identification of microRNAs that mediate the actions of EE highlights new therapeutic strategies for mental health conditions and psychiatric disease.

Intra-amygdala microinjection of TNF-α impairs the auditory fear conditioning of rats via glutamate toxicity.

During an inflammatory or infectious process, innate immune cells produce large amount of pro-inflammatory cytokines that act on the brain to cause cognitive dysfunctions. Tumor necrosis factor alpha (TNF-α) is one of the main pro-inflammatory cytokines. Thus, it is important to study how the excessive TNF-α affects the cognitive functions of central nervous system and possible antagonists to its effects. In the present study, we conducted behavioral experiments of rats to determine whether murine TNF-α administered directly into the brain would elicit behavioral effects related to learning and memory impairments. Rats subjected to single-dose intra-amygdala TNF-α infusion showed a significant delay in the acquisition and extinction of auditory fear conditioning. Accordingly, the glutamate level of the tissue samples from amygdala was elevated after the TNF-α treatment. Furthermore, pharmacological blockade of NMDAR before the TNF-α treatment reversed the TNF-α induced impairments in fear learning. Our findings suggest that TNF-α can impair the learning and memory functions through glutamate-NMDAR neurotoxicity, and present the possibility to develop therapeutic modalities directing at glutamate transmission for the treatment of neuro-inflammative dysfunctions.

Intra-amygdala microinfusion of IL-6 impairs the auditory fear conditioning of rats via JAK/STAT activation.

Though accumulating literature implicates that cytokines are involved in the pathophysiology of mental disorders, the role of interleukin-6 (IL-6) in learning and memory functions remains unresolved. The present study was undertaken to investigate the effect of IL-6 on amygdala-dependent fear learning. Adult Wistar rats were used along with the auditory fear conditioning test and pharmacological techniques. The data showed that infusions of IL-6, aimed at the amygdala, dose-dependently impaired the acquisition and extinction of conditioned fear. In addition, the results in the Western blot analysis confirmed that JAK/STAT was temporally activated-phosphorylated by the IL-6 treatment. Moreover, the rats were treated with JSI-124, a JAK/STAT3 inhibitor, prior to the IL-6 treatment showed a significant decrease in the IL-6 induced impairments of fear conditioning. Taken together, our results demonstrate that the learning behavior of rats in the auditory fear conditioning could be modulated by IL-6 via the amygdala. Furthermore, the JAK/STAT3 activation in the amygdala seemed to play a role in the IL-6 mediated behavioral alterations of rats in auditory fear learning.

Dexamethasone induced changes of neural activity in the auditory cortex of rats.

Previous studies have suggested that elevation of glucocorticoid level can alter auditory processing and may have relevance for auditory hallucinations. However, the neural mechanism underlying the glucocorticoid induced sensory change remains unclear. To examine the effects of glucocorticoid on the neuronal spike activities of sensory cortex, we topically applied dexamethasone (DEX), a glucocorticoid receptor agonist, to the auditory cortical surface of rats while recording single-unit extracellular spike activities in response to sound stimuli. Our major findings are: (1) a topical administration of DEX increased the cortical neural responses to pure-tone stimuli from 10 to 60 min after administration, while the peak response enhancement occurred at 20-30 min; (2) DEX not only markedly increased the magnitude of tone-evoked responses, but also extended the response duration and the frequency range of the neural responses; (3) the enhancement of neural responses was more salient at the higher frequency band; (4) the ratio of spontaneous firing rate between post- and pre-administration was negatively correlated with the unit's spontaneous firing rate before treatment. Our data confirm that DEX can modulate the neural activity at the cortical level and provide more information for understanding the mechanism of glucocorticoid-induced alterations in auditory processing.

Microbial communities and greenhouse gas emissions associated with the biodegradation of specified risk material in compost.

Provided that infectious prions (PrP(Sc)) are inactivated, composting of specified risk material (SRM) may be a viable alternative to rendering and landfilling. In this study, bacterial and fungal communities as well as greenhouse gas emissions associated with the degradation of SRM were examined in laboratory composters over two 14 day composting cycles. Chicken feathers were mixed into compost to enrich for microbial communities involved in the degradation of keratin and other recalcitrant proteins such as prions. Feathers altered the composition of bacterial and fungal communities primarily during the first cycle. The bacterial genera Saccharomonospora, Thermobifida, Thermoactinomycetaceae, Thiohalospira, Pseudomonas, Actinomadura, and Enterobacter, and the fungal genera Dothideomycetes, Cladosporium, Chaetomium, and Trichaptum were identified as candidates involved in SRM degradation. Feathers increased (P<0.05) headspace concentrations of CH4 primarily during the early stages of the first cycle and N2O during the second. Although inclusion of feathers in compost increases greenhouse gas emissions, it may promote the establishment of microbial communities that are more adept at degrading SRM and recalcitrant proteins such as keratin and PrP(Sc).

Optimized expression of dual reporter genes in transient transfection of purified Toxoplasma gondii using different promoters.

Fluorescent protein and luciferase genes are valuable reporter genes and have been widely used for noninvasive monitoring of gene expression in living tissues and cells. We tested expression of the dual reporter genes in transient transfection of purified Toxoplasma gondii tachyzoites. Two copies of the enhanced yellow fluorescent protein (EYFP) gene were put under the control of 3 representative T. gondii promoters (GRA1, SAG1, and DHFR). Fluorescence from each EYFP reporter was significantly higher than that from a green fluorescent protein (GFP) reporter. The GRA1-EYFP reporter gave the highest fluorescence. Although both fluorescence and luciferase were expressed in the dual reporter system, the luciferase reporter was more efficient than either the EYFP or GFP reporters, and it required fewer parasites to be successfully used.

Expression of the tandem enhanced yellow fluorescent marker gene in Toxoplasma gondii.

Fluorescent protein is a useful tool for monitoring gene expression and studying biological processes of organisms including parasites. To improve the transfection efficiency and fluorescent intensity in Toxoplasma gondii, a new transient expression vector, RGES, containing the tandem enhanced yellow fluorescent protein gene, EYFP-EYFP, under the control of the parasite dense granule protein 1 (GRA1) promoter was constructed. The RGES plasmid was introduced into T. gondii RH strain by electroporation. A high proportion of tachyzoites (approximately 25.5%) was transfected with the EYFP-EYFP gene and high fluorescence intensity was detected 48-72 h after electroporation. The tandem EYFP gene and its expression in the transfectants were confirmed by polymerase chain reaction (PCR), reverse transcriptase PCR, and Western blotting. The efficiency of transfection and fluorescent intensity of EYFP-EYFP were greater than that of the single EYFP or green fluorescent protein. The EYFP-EYFP gene is a better marker for the study of biological processes of T. gondii.

Development of a rapid real-time PCR assay for detection and quantification of four familiar species of Chlamydiaceae.

Chlamydiae are one of the causative agents of various diseases in animals and human beings, which include abortion, pneumonia, gastroenteritis, encephalomyelitis, conjunctivitis, arthritis and sexually transmitted diseases. Much work has been carried out to attempt to develop an efficient pathogen detection strategy. Here, we presented a Chlamydiaceae-specific 23S rRNA-based real-time PCR assay for simultaneous detection and quantification of four members of Chlamydiaceae family, C. trachomatis, C. psittaci, C. pneumoniae and C. pecorum, using SYBR Green and Lightcycler. The assay was characterized using plasmid constructs of the bacteria and verified on standard strains of all four species of the Chlamydiaceae and a large cohort of clinical samples collected from human and animals by comparison with fluorescence immunohistochemistry method. The results showed that the present real-time PCR assay was of high specificity and sensitivity. It was capable of detecting as few as 250fg of chlamydial DNA (equivalent to 10(-1)IFU) and was applicable to both liquid cultures and clinical samples. This assay may therefore offer a rapid, economic and reliable means for screening of the chlamydiaceae pathogens.

Pathology of guinea pigs experimentally infected with a novel reovirus and coronavirus isolated from SARS patients.

Guinea pigs were inoculated with a reovirus (ReoV) and coronavirus (SARS-CoV) isolated from SARS patients to determine their potential role in the etiology of SARS. Animals infected with ReoV died between day 22 and day 30 postinoculation (PI) while 70% of the animals inoculated with ReoV and SARS-CoV died between day 4 to day 7 PI. The titer of neutralizing antibodies against ReoV and SARS-CoV ranged from 80 to 160 when the animals were inoculated with the two viruses, respectively, while the titer of the antibodies was just below 10 in coinfections. The animal inoculated with ReoV developed diffuse alveolar damage similar to the exudative and leakage inflammation found in SARS patients, and was characterized by diffuse hemorrhage, fibroid exudation, hyaline membrane formation, and type II pneumocytes hyperplasia in alveolar interstitia. The pulmonary epithelial necrosis, excoriation, and early fibrosis of pulmonary tissue were only observed in ReoV-SARS-CoV groups and in SARS-CoV/ReoV groups. Other typical pathological changes included hemorrhagic necrosis in lymph nodes and spleen and hydropic degeneration in the liver. On the contrary, guinea pigs infected with SARS-CoV only developed interstitial pneumonitis. Our experiment demonstrate that ReoV might be one of the primary causes of SARS, since simultaneous coinfection can duplicate the typical pathological changes similar to that of SARS patients. This guinea pig model may provide a useful animal model for SARS.

Comparative analysis of the prion protein open reading frame nucleotide sequences in peacock and parakeet.

The open reading frame of peacock and parakeet prion protein (PrP) genes was cloned and sequenced. The peacock and parakeet PrP genes consisted of 833 and 866 nucleotides encoding 266 and 277 amino acids, respectively (GenBank Accession numbers AY365065 and AY365066). Sequence analysis showed that the peacock and parakeet PrP genes had 93.67% homology to each other, 94.04% and 99.64% homology to the chicken PrP gene and 46.0% and 42.1% similarity to the mammalian PrP genes, respectively. The structural features of all known mammalian and avian PrPs, including N-terminal signal peptides, tandem repeats, conserved hydrophobic region, disulfide bridges and glycoinositol phospholipid anchor, were also found in peacock and parakeet PrPs. The parakeet and peacock PrPs, however, differed in the hexarepeat region, with the peacock having six and the parakeet having seven hexarepeats. The phylogenetic analysis showed that the PrP genes in 52 species were clustered into 2 distinct lineages, the avian and the mammalian. The peacock and parakeet PrP genes belonged to the same lineage but the peacock PrP was sub-classed with the pigeon PrP and the parakeet PrP was sub-classed with the duck and chicken PrPs. The present work added two more species data to the collection of the PrP genes and supported the previous findings that the PrP genes are highly conserved across species.