Somatostatin slows Aß plaque deposition in aged APPNL-F/NL-F mice by blocking Aß aggregation.

The neuroendocrine peptide somatostatin (SST) has long been thought of as influencing the deposition of the amyloid ß peptide (Aß) in Alzheimer's disease (AD). Missing have been in vivo data in a relevant Aß amyloidosis model. Here we crossed AppNL-F/NL-F mice with Sst-deficient mice to assess if and how the presence of Sst influences pathological hallmarks of Aß amyloidosis. We found that Sst had no influence on whole brain neprilysin transcript, protein or activity levels, an observation that cannot be accounted for by a compensatory upregulation of the Sst paralog, cortistatin (Cort), that we observed in 15-month-old Sst-deficient mice. Sst-deficiency led to a subtle but significant increase in the density of cortical Aß amyloid plaques. Follow-on western blot analyses of whole brain extracts indicated that Sst interferes with early steps of Aß assembly that manifest in the appearance of SDS-stable smears of 55-150 kDa in Sst null brain samples. As expected, no effect of Sst on tau steady-state levels or its phosphorylation were observed. Results from this study are easier reconciled with an emerging body of data that point toward Sst affecting Aß amyloid plaque formation through direct interference with Aß aggregation rather than through its effects on neprilysin expression.

Soil pH determines arsenic-related functional gene and bacterial diversity in natural forests on the Taibai Mountain.

Arsenic-related functional genes are ubiquitous in microbes, and their distribution and abundance are influenced by edaphic factors. In arsenic-contaminated soils, soil arsenic content and pH determine the distribution of arsenic metabolizing microorganisms. In the uncontaminated natural ecosystems, however, it remains understudied for the key variable factor in determining the variation of bacterial assembly and mediating the arsenic biogeographical cycles. Here, we selected natural forest soils from southern and northern slopes along the altitudinal gradient of Taibai Mountain, China. The arsenic-related functional genes and soil bacterial community was examined using GeoChip 5.0 and high-throughput sequencing of 16S rRNA genes, respectively. It was found that arsenic-related functional genes were ubiquitous in tested forest soils. The gene arsB has the highest relative abundance, followed by arsC, aoxB, arrA, arsM, and arxA. The arsenic-related functional genes distribution on two slopes were decoupled from their corresponding bacterial community. Though there are higher abundance of bacterial communities on the northern slope than that on the southern slope, for arsenic-related functional genes, the abundance has the contrary trend which showing the more arsenic-related functional genes on the southern slope. In the top ten phyla, Proteobacteria and Actinobacteria were dominant phyla which affected the abundance of arsenic-related functional genes. Redundancy analysis and variance partitioning analysis indicated that soil pH, organic matter and altitude jointly determined the arsenic-related functional genes diversity in the two slopes of Taibai Mountain, and soil pH was a key factor. This indicates that the lower pH may shape more microbes with arsenic metabolic capacity. These findings suggested that soil pH plays a significant role in regulating the distribution of arsenic-related functional microorganisms, even for a forest ecosystem with an altitudinal gradient, and remind us the importance of pH in microbe mediated arsenic transformation.

Oocytes could rearrange immunoglobulin production to survive over adverse environmental stimuli.

Immunoglobulins are key humoral immune molecules produced and secreted by B lymphocytes at various stages of differentiation. No research has reported whether immunoglobulins are present in the non-proliferative female germ cells-oocytes-and whether they are functionally important for oocyte quality, self-protection, and survival. Herein, we found that IgG was present in the oocytes of immunodeficient mice; the IgG-VDJ regions were highly variable between different oocytes, and H3K27Ac bound and regulated the IgG promoter region. Next, IgG mRNA and protein levels increased in response to LPS, and this increment was mediated by CR2 on the oocyte membrane. Finally, we revealed three aspects of the functional relevance of oocyte IgG: first, oocytes could upregulate IgG to counteract the increased ROS level induced by CSF1; second, oocytes could upregulate IgG in response to injected virus ssRNA to maintain mitochondrial integrity; third, upon bacterial infection, oocytes could secrete IgG, subsequently encompassing the bacteria, thus increasing survival compared to somatic cells. This study reveals for the first time that the female germ cells, oocytes, can independently adjust intrinsic IgG production to survive in adverse environments.

Cardiac glycoside-mediated turnover of Na, K-ATPases as a rational approach to reducing cell surface levels of the cellular prion protein.

It is widely anticipated that a reduction of brain levels of the cellular prion protein (PrPC) can prolong survival in a group of neurodegenerative diseases known as prion diseases. To date, efforts to decrease steady-state PrPC levels by targeting this protein directly with small molecule drug-like compounds have largely been unsuccessful. Recently, we reported Na,K-ATPases to reside in immediate proximity to PrPC in the brain, unlocking an opportunity for an indirect PrPC targeting approach that capitalizes on the availability of potent cardiac glycosides (CGs). Here, we report that exposure of human co-cultures of neurons and astrocytes to non-toxic nanomolar levels of CGs causes profound reductions in PrPC levels. The mechanism of action underpinning this outcome relies primarily on a subset of CGs engaging the ATP1A1 isoform, one of three α subunits of Na,K-ATPases expressed in brain cells. Upon CG docking to ATP1A1, the ligand receptor complex, and PrPC along with it, is internalized by the cell. Subsequently, PrPC is channeled to the lysosomal compartment where it is digested in a manner that can be rescued by silencing the cysteine protease cathepsin B. These data signify that the repurposing of CGs may be beneficial for the treatment of prion disorders.

Dynamic changes in the phenolic composition and antioxidant activity of oats during simultaneous hydrolysis and fermentation.

Solid-state fermentation (SSF) is the preferred method of enhancing the phenolic content of oats, while scientific optimization for improving specific phenolic compounds is limited. In this study, sequential targeting of phenolic conversion in simultaneous hydrolysis and fermentation (SHF) of oats was investigated. The results revealed that SHF with adding cellulase at 0, 6 and 12 days could increase the total phenolic content by 4.4%, 67.8% and 59.1%, respectively, over that of SSF. The α-amylase and CMCase activity were highly correlated with the soluble and insoluble phenolic contents in SHF (-6 and -12) systems (r > 0.8, p < 0.05). Interestingly, the content of phenolic fraction, such as ferulic acid, was up-regulated, whereas sinapic acid was down-regulated. These results indicated that the phenolic conversion occurred in SHF, resulting in variation in DPPH and ABTS+ radical scavenging abilities. This research provided metabolic understanding of the optimization of phenolic compounds to increase the functional ingredient of oats.

X-chromosome genetic association test incorporating X-chromosome inactivation and imprinting effects.

Studies have shown that many complex diseases are sex-determined. When conducting genetic association studies on X-chromosome, there are two important epigenetic factors which should be considered simultaneously: X-chromosome inactivation and genomic imprinting. Currently, there have been several association tests accounting for the information on X-chromosome inactivation. However, these tests do not take the imprinting effects into account. In this paper, we propose a novel association test simultaneously incorporating X-chromosome inactivation and imprinting effects based on case-parent trios and control-parent trios for female offspring and case-control data for male offspring, denoted by MLRXCII. Extensive simulation studies are carried out to investigate the type I error rate and the test power of the proposed MLRXCII . Simulation results demonstrate that the proposed test controls the type I error rate well andis more powerful than the existing method when imprinting effects exist. The proposed MLRXCII test is valid and powerful in genetic association studies on X-chromosome for qualitative traits and thus is recommended in practice.

Galectin-3 as a candidate upstream biomarker for quantifying risks of myocardial ageing.

AIMS: Galectin-3 (Gal-3) is implicated in the pathogenesis of heart failure and is also influenced by ageing. This study aims to determine the extent to which Gal-3 levels estimate odds of myocardial dysfunction in ageing cohorts, 'upstream' prior to clinical disease. METHODS AND RESULTS: Four hundred seventy-five asymptomatic subjects underwent simultaneous assessments of cardiovascular structure and function, with measurements of circulating Gal-3. Myocardial dysfunction was defined as impaired myocardial relaxation (ratio of peak velocity flow in early diastole E (m/s) to peak velocity flow in late diastole by atrial contraction A (m/s) <0.84) (mean E/A ratio 0.84 in the cohort). Of 475 subjects (mean age 68 ± 12 years, 231 women), 222 (47%) had myocardial dysfunction. Subjects with myocardial dysfunction were older (mean age 73 ± 5 vs. 64 ± 14 years, P < 0.0001), and more had hypertension (59 vs. 40%, P < 0.0001), dyslipidaemia (54 vs. 39%, P = 0.001), diabetes mellitus (25 vs. 14%, P = 0.002), higher body mass index (BMI) (24 vs. 23 kg/m2 , P = 0.002), and higher heart rate (76 vs. 71 b.p.m., P = 0.0001). Participants with impaired myocardial relaxation had lower peak velocity flow in early diastole E (0.6 ± 0.1 vs. 0.8 ± 0.2 m/s, P < 0.0001), higher peak velocity flow in late diastole by atrial contraction A (0.9 ± 0.1 vs. 0.7 ± 0.2 m/s, P < 0.0001), and higher mitral valve flow deceleration time (224.7 ± 43.2 vs. 204.8 ± 33.1 m/s, P < 0.0001). Participants with impaired myocardial relaxation had higher Gal-3 levels (17.2 ± 6.2 vs. 15.5 ± 4.1, P = 0.0004) but similar B-type natriuretic peptide (37 ± 4 vs. 34 ± 29, P = 0.37) and high-sensitivity troponin I (21 ± 72 vs. 11 ± 41, P = 0.061) levels and urine microalbumin-to-creatinine ratio (4.6 ± 8.1 vs. 4.2 ± 10.8, P = 0.75) compared with those without impaired myocardial relaxation. After multivariable adjustments, Gal-3 [odds ratio (OR) 1.05, 95% confidence interval (CI) 1.00-1.10, P = 0.039], age (OR 2.60, 95% CI 1.64-4.11, P < 0.0001), BMI (OR 2.16, 95% CI 1.44-3.23, P < 0.0001), and heart rate (OR 1.04, 95% CI 1.02-1.06, P < 0.0001) were associated with impaired myocardial relaxation. Adjusted ORs (95% CI) for myocardial dysfunction were 1.0 (ref), 1.62 (0.92-2.85), 1.92 (1.08-3.41), and 2.01 (1.11-3.66) across consecutive quartiles of Gal-3 after adjustment for age, BMI, risk factors, and heart rate. CONCLUSIONS: Among asymptomatic community-dwelling elderly adults, the highest quartile of Gal-3 was associated with two-fold increased odds of myocardial dysfunction compared with the lowest quartile of Gal-3. Gal-3 may have a role as an 'upstream' biomarker in estimating odds of myocardial ageing prior to clinical disease.

A statistical measure for the skewness of X chromosome inactivation based on case-control design.

BACKGROUND: Skewed X chromosome inactivation (XCI), which is a non-random process, is frequently observed in both healthy and affected females. Furthermore, skewed XCI has been reported to be related to many X-linked diseases. However, no statistical method is available in the literature to measure the degree of the skewness of XCI for case-control design. Therefore, it is necessary to develop methods for such a task. RESULTS: In this article, we first proposed a statistical measure for the degree of XCI skewing by using a case-control design, which is a ratio of two logistic regression coefficients after a simple reparameterization. Based on the point estimate of the ratio, we further developed three types of confidence intervals (the likelihood ratio, Fieller's and delta methods) to evaluate its variation. Simulation results demonstrated that the likelihood ratio method and the Fieller's method have more accurate coverage probability and more balanced tail errors than the delta method. We also applied these proposed methods to analyze the Graves' disease data for their practical use and found that rs3827440 probably undergoes a skewed XCI pattern with 68.7% of cells in heterozygous females having the risk allele T active, while the other 31.3% of cells keeping the normal allele C active. CONCLUSIONS: For practical application, we suggest using the Fieller's method in large samples due to the non-iterative computation procedure and using the LR method otherwise for its robustness despite its slightly heavy computational burden.

A robust and powerful test for case-control genetic association study on X chromosome.

Hundreds of genome-wide association studies were conducted to map the disease genes on autosomes in human beings. It is known that many complex diseases are sex-determined and X chromosome is expected to play an important role. However, only a few single-nucleotide polymorphisms on X chromosome were found to be significantly associated with the diseases under study. On the other hand, to balance the genetic effect between two sexes, X chromosome inactivation occurs in most of X-linked genes by silencing one copy of two X chromosomes in females and dosage compensation is achieved. A few association studies on X chromosome incorporated the information on dosage compensation. However, some of them require the assumption of Hardy-Weinberg equilibrium and some need to specify the underlying genetic model. Therefore, in this article, we propose a novel method for association by taking account of different dosage compensation patterns. The proposed test is a robust approach because it requires neither specifying the underlying genetic models nor the assumption of Hardy-Weinberg equilibrium. Further, the proposed method allows for different deviations from Hardy-Weinberg equilibrium between cases and controls. Simulation results demonstrate that our proposed method generally outperforms the existing methods in terms of controlling the size and the test power. Finally, we apply the proposed test to the meta-analysis of the Graves' disease data for its practical use.

N-Terminal pro C-Type Natriuretic Peptide (NTproCNP) and myocardial function in ageing.

Ageing-related alterations in cardiovascular structure and function are commonly associated with chronic inflammation. A potential blood-based biomarker indicative of a chronic inflammatory state is N-Terminal Pro C-Type Natriuretic Peptide (NTproCNP). We aim to investigate associations between NTproCNP and ageing-related impairments in cardiovascular function. Community-based participants underwent same-day assessment of cardiovascular function and circulating profiles of plasma NTproCNP. Associations between cardiovascular and biomarker profiles were studied in adjusted models including standard covariates. We studied 93 participants (mean age 73 ± 5.3 years, 36 women), of whom 55 (59%) had impaired myocardial relaxation (ratio of peak velocity flow in early diastole E (m/s) to peak velocity flow in late diastole by atrial contraction A (m/s) <0.84). Participants with impaired myocardial relaxation were also found to have lower peak early phase filling velocity (0.6 ± 0.1 vs 0.7 ± 0.1, p < 0.0001) and higher peak atrial phase filling velocity (0.9 ± 0.1 vs 0.7 ± 0.1, p < 0.0001). NTproCNP levelswere significantly lower among participants with impaired myocardial relaxation (16.4% vs 39.5% with NTproCNP ≥ 19, p = 0.012). After multivariable adjustments, NTproCNP was independently associated with impaired myocardial relaxation (OR 2.99, 95%CI 1.12-8.01, p = 0.029). Community elderly adults with myocardial ageing have lower NTproCNP levels compared to those with preserved myocardial function. Given that impaired myocardial relaxation probably represents early changes within the myocardium with ageing, NTproCNP may be useful as an 'upstream' biomarker useful for charting myocardial ageing.

Effect of the 2008 cold spell on preterm births in two subtropical cities of Guangdong Province, Southern China.

BACKGROUND: A few studies have reported that low temperatures were associated with an increased risk of preterm birth. However, the effect of extreme weather events, such as cold spell, on preterm birth has not been studied in China. OBJECTIVE: This study was conducted to evaluate the impact of the 2008 cold spell on preterm birth in two subtropical cities of Guangdong Province. METHODS: Data of daily preterm birth, air pollution and meteorological variables from 2006 to 2010 were collected in Dongguan and Shenzhen. A Poisson regression with a distributed lag nonlinear model was used to investigate the association between the 2008 cold spell and daily rate of preterm birth for each city. RESULTS: During the 2008 cold spell, total vaginal preterm births were increased by 22.44% and 21.25% in Dongguan and Shenzhen, respectively. The effect of the cold spell on preterm births lasted for more than 1 week, the lag0-6 days cumulative relative risk (RR) is 1.32 (95% CI: 1.10-1.58) and 1.40 (95% CI: 1.18-1.68) in Dongguan and Shenzhen, respectively. The effects were found to be more pronounced for the pregnant women with 34-36 gestation weeks, maternal age < 35 years group. CONCLUSION: This study demonstrates that cold spell could increase the risk of preterm births in Dongguan and Shenzhen, and the effect lasts for more than 1 week. Specific measures should be considered to protect the pregnant women, especially the vulnerable subgroups.

Variations in Monascus pigment characteristics and biosynthetic gene expression using resting cell culture systems combined with extractive fermentation.

Monascus pigments are promising sources of natural food colorants, and their productivity can be improved by a novel extractive fermentation technology. In this study, we investigated the variations in pigment characteristics and biosynthetic gene expression levels in resting cell culture systems combined with extractive fermentation in Monascus anka GIM 3.592. Although the biomass was low at about 6 g/L DCW, high pigment titer of approximately 130 AU470 was obtained in the resting culture with cells from extractive fermentation, illustrating that it had a good biocatalytic activity for pigment synthesis. The oxidation-reduction potential value correlated with the rate of relative content of the intracellular orange pigments to the yellow pigments (O/Y, r > 0.90, p < 0.05), indicating that the change in pigment characteristics may be responsible for the cellular redox activity. The up- or down-regulation of the pigment biosynthetic genes (MpFasA2, MpFasB2, MpPKS5, mppD, mppB, mppR1, and mppR2) in the resting culture with extractive culture cells was demonstrated by real-time quantitative polymerase chain reaction analysis. Moreover, the mppE gene associated with the yellow pigment biosynthesis was significantly (p < 0.05) down-regulated by about 18.6%, whereas the mppC gene corresponding to orange pigment biosynthesis was significantly (p < 0.05) up-regulated by approximately 21.0%. These findings indicated that extractive fermentation was beneficial for the biosynthesis of the intracellular orange pigment. The mechanism described in this study proposes a potential method for the highly efficient production of Monascus pigments.

Enzymatic action mechanism of phenolic mobilization in oats (Avena sativa L.) during solid-state fermentation with Monascus anka.

This work aims to investigate the effects of carbohydrate-hydrolysing enzymes on the release of phenolics in oat fermentation with Monascus anka. There were good correlations between phenolic content and α-amylase, xylanase and FPase activities. A high level of α-amylase activity (141.07 U/g) was observed, while xylanase (2.40 U/g), total cellulase (0.52 U/g) and ß-glucosidase activities (0.028 U/g) were relatively low in the fermentation system. The phenolic content of oat powder treated with crude enzyme from fermented oats significantly increased, especially that of the ferulic acid in the insoluble fraction and the vanillic acid in the soluble fraction. The surface SEM morphology of the oats showed that the cell wall structure was damaged by the crude enzyme treatment, which led to the release of phenolics. This study could provide metabolic understanding for optimization of phenolic compounds which could more efficiently increase the nutrition of oat intended for functional food ingredients.

Tracking of pigment accumulation and secretion in extractive fermentation of Monascus anka GIM 3.592.

BACKGROUND: Monascus pigments are promising sources for food and medicine due to their natural food-coloring functions and pharmaceutical values. The innovative technology of extractive fermentation is used to promote pigment productivity, but reports of pigment trans-membrane secretion mechanism are rare. In this study, tracking of pigment accumulation and secretion in extractive fermentation of Monascus anka GIM 3.592 was investigated. RESULTS: The increased vacuole size in mycelia correlated with fluorescence intensity (r > 0.85, p < 0.05), which indicates that intracellular pigments with strong fluorescence accumulated in the cytoplasmic vacuole. After adding nonionic surfactant Triton X-100, the uptake of rhodamine123 (Rh123) and 1-N-phenylnaphthylamine (NPN) and the release of K+ and Na+ rapidly increased, demonstrating that the physiological performances of the cell membrane varied upon damaging the integrity, increasing the permeability, and changing the potential. Simultaneously, the fatty acid composition also varied, which caused a weak fluidity in the membrane lipids. Therefore, the intracellular pigments embedded in Triton X-100 were secreted through the ion channels of the cell membrane. Dense, spherical pigment-surfactant micelles with an average size of 21 nm were distributed uniformly in the extraction broth. Based on the different pigment components between extractive fermentation and batch fermentation, a threefold decrease in the NAD+/NADH ratio in mycelia and a more than 200-fold increase in glucose-6-phosphate dehydrogenase (G6PDH) activity in extracellular broth occurred, further suggesting that a reduction reaction for pigment conversion from orange pigments to yellow pigments occurred in non-aqueous phase solution. CONCLUSIONS: A putative model was established to track the localization of Monascus pigment accumulation and its trans-membrane secretion in extractive fermentation. This finding provides a theoretical explanation for microbial extractive fermentation of Monascus pigments, as well as other non-water-soluble products.

Fingerprint profiles of flavonoid compounds from different Psidium guajava leaves and their antioxidant activities.

Flavonoids are the main active components in Psidium guajava leaves and have many multi-physiological functions. In this study, the flavonoid compositions were identified in the Psidium guajava leaves samples using a high-performance liquid chromatography with time-of-flight electrospray ionization mass spectrometry method. A high-performance liquid chromatography fingerprint method, combined with chemometrics, was used to perform a quality assessment of the Psidium guajava leaves samples. The eight identified flavonoid compounds including rutin, isoquercitrin, quercetin-3-O-ß-d-xylopyranoside, quercetin-3-O-α-l-arabinopyranoside, avicularin, quercitrin, quercetin, and kaempferol were used as the chemical markers. The antioxidant activity of 15 batches of samples was examined using three different methods, and the results revealed the Psidium guajava leaves samples that had higher contents of the flavonoid compounds, glycoside and aglycone, possessed the highest antioxidant capacities. Consequently, a combination of chromatographic fingerprints and chemometric analyses was used for a quality assessment of Psidium guajava leaf tea and its derived products, which can lay the foundation for the development of plant tea resources or other herbs.

Saturation effect and transmembrane conversion of Monascus pigment in nonionic surfactant aqueous solution.

Extractive fermentation in a nonionic surfactant aqueous solution provides a promising and efficient method to produce Monascus pigments. The behaviour of pigment secretion during the extractive cultivation was investigated in the present work. The results revealed that the secretion of intracellular pigment was limited by its saturation concentration in the nonionic surfactant aqueous solution. The intracellular pigment was completely extracted to the outside of the cell at a low cell density and high concentration of Triton X-100 (TX) in fermentation broth; otherwise, a restriction for pigment extraction would occur. The decrement of the intracellular orange and yellow pigments was inconsistent with the increment of extracellular pigments with an increase in the TX concentration. It could be inferred that the intracellular orange pigment was converted to extracellular yellow pigment during the transmembrane secretion process, which might be attributed to the enzyme catalysis in the non-aqueous phase solution. This study helps explain the mechanism of variation of pigment characteristic and extraction capacity in extractive fermentation.

[Hsa-miR-206 inhibits the migration and invasion of breast cancer by targeting Cx43].

OBJECTIVE: To explore the effects of hsa-miR-206 on the proliferation, migration and invasion of breast cancer. METHODS: The hsa-miR-206 mimics, inhibitors and their paired negative controls were transfected into human breast cancer cell line MDA-MB-231 by liposome. The proliferation of cell was evaluated by CCK-8 and the migration and invasion was detected by Transwell. Matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 9 (MMP-9), breast cancer metastasis-suppressor 1 (BRMS-1) and connexin 43 (Cx43) were detected by both quantitative polymerase chain reaction (qPCR) and Western blot. The expression of miR-206 was detected by qPCR. Dual luciferase assay was detected to confirm the specific binding sites of miR-206 and Cx43. RESULTS: (1) The proliferation activity of 206m-group cell (0.74 ± 0.16) was significantly lower than that of control group (1.12 ± 0.23) (t = -3.066, P = 0.037) while that of 206i-group cell (1.43 ± 0.26) was higher than that of control group (0.98 ± 0.14) (t = 3.635, P = 0.022). (2) Transwell tests showed the migration and invasion of 206m-group cell decreased significantly (migration:0.56 ± 0.01 vs 0.63 ± 0.01, t = -23.00, P = 0.002; invasion:0.79 ± 0.01 vs 0.99 ± 0.01, t = -21.200, P = 0.002), but that of 206i-group cell increased significantly (migration:0.97 ± 0.11 vs 0.61 ± 0.09, t = 32.787, P = 0.001; invasion:1.10 ± 0.01 vs 0.93 ± 0.05, t = 5.167, P = 0.035). (3) The expressions of MMP-2,MMP-9 and Cx43 decreased and the expression of BRMS-1 increased in 206m-group cell and vice versa in 206i group. (4) The expression of miR-206 in lymph node-negative group of clinical breast cancer sample was higher than that of lymph node-positive one. And there was statistical difference (Z = -2.098, P = 0.003). And the expression of Cx43 was opposite. (5) Dual luciferase reporter assay confirmed the specific binding sites of hsa-miR-206 and Cx43. CONCLUSION: Hsa-miR-206 has negative controls of proliferation, migration and invasion of breast cancer cell by targeting Cx43.