Effect of Sunflower Kernel Peptides Produced by Dual-Degradation on the Growth Performance, Nutrient Digestibility, and Health Status of Broilers

The purpose of this study was to investigate the effect of sunflower kernel peptides produced by enzymatic digestion, fermentation, or both on the growth performance, nutrient digestibility, and health status of broilers. Four diets contained 20% of sunflower kernel meal as its raw form (CON) or degraded by protease (ESM), Bacillus pumilus (FSM), or both (DSM). A total of 480 yellow broilers at one day old were randomly distributed to 4 groups with 6 replicates of 20 chicks each. The feeding trial lasted for 63 d. Results showed that peptide content was increased (p<0.001) from 3.97% (CON) to 32.5% (ESM), 24.2% (FSM), and 39.1% (DSM). The three sunflower peptide groups improved (p≤0.001) feed intake and body weight gain. The peptide groups increased (p≤0.015) ileal apparent digestibility of dry matter, energy, crude protein, and amino acids (methionine, lysine, tryptophan, and threonine). Furthermore, the peptide groups improved (p≤0.029) the health status by increasing serum immunoglobulins (IgA, IgG) and glutathione peroxidase. Additionally, among the peptide groups, DSM showed more pronounced effects (p<0.05) on these parameters than ESM or FSM. It is concluded that dual-degradation by enzymolysis and fermentation has a better improvement in the nutrition and application of sunflower kernel meal in broilers.(AU)

Probiotic Lactobacilli Improved Growth Performance and Attenuated Salmonella Typhimurium Infection Via Jak/Stat Signaling in Broilers

This study aimed to investigate the effect of Lactobacillus plantarum DPP8 and Lactobacillus acidophilus C7282 in feed supplementation on growth performance, Salmonella invasion, inflammation, and mediating signaling in broilers infected with Salmonella Typhimurium (S. Typhimurium). A total of 240 broilers at day old were randomly allocated into four groups, orally infected with S. Typhimurium and supplemented with individual or combined Lactobacilli DPP8 and C7282 at doses of 0 (control), 1010 (individual), or 2.0 × 1010 (combination) cfu/kg of diet for 21 d. The results showed that supplementing Lactobacilli improved (p 0.05) feed intake and body weight gain and decreased (p 0.05) S. Typhimurium load in the caecum, harder gland, spleen and bursa of Fabricius. Also, the supplements decreased (p 0.05) interleukin (1/2/4), tumor necrosis factor and interferon in the serum, enhanced (p 0.05) interleukin 10, and downregulated gene expressions of inflammatory mediators including Janus kinase (Jak2/3), signal transducer and activator of transcription protein (STAT3/4/5/6) in the intestinal mucosa. In contrast, diets containing DPP8 exhibited greater effects on the inhibition of the pathogen and inflammatory response than C7282. The obtained data suggest that Lactobacilli C7282 and DPP8 can be used as feed additives to inhibit colonization and translocation of S. Typhimurium and inflammatory responses via downregulating Jak/STAT signaling in broilers.(AU)

Probiotic Lactobacilli Improved Growth Performance and Attenuated Salmonella Typhimurium Infection Via Jak/Stat Signaling in Broilers

This study aimed to investigate the effect of Lactobacillus plantarum DPP8 and Lactobacillus acidophilus C7282 in feed supplementation on growth performance, Salmonella invasion, inflammation, and mediating signaling in broilers infected with Salmonella Typhimurium (S. Typhimurium). A total of 240 broilers at day old were randomly allocated into four groups, orally infected with S. Typhimurium and supplemented with individual or combined Lactobacilli DPP8 and C7282 at doses of 0 (control), 1010 (individual), or 2.0 × 1010 (combination) cfu/kg of diet for 21 d. The results showed that supplementing Lactobacilli improved (p 0.05) feed intake and body weight gain and decreased (p 0.05) S. Typhimurium load in the caecum, harder gland, spleen and bursa of Fabricius. Also, the supplements decreased (p 0.05) interleukin (1/2/4), tumor necrosis factor and interferon in the serum, enhanced (p 0.05) interleukin 10, and downregulated gene expressions of inflammatory mediators including Janus kinase (Jak2/3), signal transducer and activator of transcription protein (STAT3/4/5/6) in the intestinal mucosa. In contrast, diets containing DPP8 exhibited greater effects on the inhibition of the pathogen and inflammatory response than C7282. The obtained data suggest that Lactobacilli C7282 and DPP8 can be used as feed additives to inhibit colonization and translocation of S. Typhimurium and inflammatory responses via downregulating Jak/STAT signaling in broilers.

Dietary Lactic Acid Bacteria Modulate Yolk Components and Cholesterol Metabolism by Hmgr Pathway in Laying Hens

This study aimed to investigate the effect of dietary lactic acid bacteria (LAB) on egg production, yolk components, cholesterol metabolism, and enterohepatic circulation of bile acids in hens. Four treatment diets included a control and LAB added at 3 × 105 (low), 3 × 107 (medium), or 3 × 109 (high) cfu/kg. The treatment LAB contained equal amounts of Lactobacillus acidophilus, Lactobacillus plantarum, and Enterococcus faecium. Results showed that high LAB increased (p 0.05) laying rate, egg mass, and yolk phospholipid, but decreased (p 0.05) yolk triglyceride and phosvitin. Diets with LAB decreased (p 0.05) yolk and serum cholesterol content, and serum bile acid by 9.3 to 39.9%. In liver, high LAB downregulated (p 0.05) mRNA expression of serine/threonine kinase 11 (STK11), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR), AMP-activated protein kinase catalytic subunit (PRKAA1, 2), and protein phosphatase catalytic subunits (PPP2CA, PPP2CB and PPP3CA) by 49.5 to 175.4%. In mucosa, high LAB downregulated (p 0.05) PRKAA1 and HMGR by 68.2 and 69.6%, respectively; but upregulated (p 0.05) PPP2CA and PPP2CB by 51.2 and 45%, respectively. Linear decreasing (p0.035) responses to LAB doses were found on cholesterol, phosvitin, bile acid, and hepatic gene expressions, and quadratic (p0.006) effects on yolk cholesterol and hepatic STK11. It is concluded that probiotic LAB can improve yolk components and decrease hepatic cholesterol synthesis by regulating HMGR pathway in hens.(AU)

Dietary Lactic Acid Bacteria Modulate Yolk Components and Cholesterol Metabolism by Hmgr Pathway in Laying Hens

This study aimed to investigate the effect of dietary lactic acid bacteria (LAB) on egg production, yolk components, cholesterol metabolism, and enterohepatic circulation of bile acids in hens. Four treatment diets included a control and LAB added at 3 × 105 (low), 3 × 107 (medium), or 3 × 109 (high) cfu/kg. The treatment LAB contained equal amounts of Lactobacillus acidophilus, Lactobacillus plantarum, and Enterococcus faecium. Results showed that high LAB increased (p 0.05) laying rate, egg mass, and yolk phospholipid, but decreased (p 0.05) yolk triglyceride and phosvitin. Diets with LAB decreased (p 0.05) yolk and serum cholesterol content, and serum bile acid by 9.3 to 39.9%. In liver, high LAB downregulated (p 0.05) mRNA expression of serine/threonine kinase 11 (STK11), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR), AMP-activated protein kinase catalytic subunit (PRKAA1, 2), and protein phosphatase catalytic subunits (PPP2CA, PPP2CB and PPP3CA) by 49.5 to 175.4%. In mucosa, high LAB downregulated (p 0.05) PRKAA1 and HMGR by 68.2 and 69.6%, respectively; but upregulated (p 0.05) PPP2CA and PPP2CB by 51.2 and 45%, respectively. Linear decreasing (p0.035) responses to LAB doses were found on cholesterol, phosvitin, bile acid, and hepatic gene expressions, and quadratic (p0.006) effects on yolk cholesterol and hepatic STK11. It is concluded that probiotic LAB can improve yolk components and decrease hepatic cholesterol synthesis by regulating HMGR pathway in hens.

Effect of broccoli residues fermented with probiotics on the growth performance and health status of broilers challenged with Clostridium perfringens

This study aimed at investigating the effects of broccoli residues fermented with probiotics (BF) on the growth performance, immunity, and gut health in broilers challenged with Clostridium perfringens (C. perfringens). A total of 600 broilers (one day old) were randomly allotted into five treatments with six replicates of 20 birds each and were reared until 42 days of age. The treatments included a positive control (PC, fed a basal diet and reared on uncontaminated litter), a negative control (NC, birds reared on litter contaminated with C. perfringens and fed a basal diet), and NC plus BF at 25, 50 or 75 g/kg of diet. The BF contained yeast 3.1 × 10 (7) cfu/g, lactic acid bacteria 9.5 × 10 (6) cfu/g and Bacillus subtilis 3.5 × 10 (6) cfu/g. Birds in the NC group showed lower (p<0.05) feed intake and body weight gain, whereas BF supplementation recovered (p<0.05) the growth performance to the levels of PC group. Dietary BF at 50and 75 g/kg reduced (p<0.05) broiler mortality. Similarly, compared to the NC group, BF increased (p<0.05) immune organ weights and serum immunoglobulins A, G, and M to the levels of PC group. The ileal populations of Escherichia coli and Gram-negative bacteria were decreased (p<0.05) by BF to the levels of PC, and C. perfringens was also decreased (p<0.05) by BF. The serum profiles of mono- and di-amine oxidase were decreased (p<0.05) by BF. BF at 75 g/kg reduced (p<0.05) monoamine oxidase compared with the other BF doses. The results suggest that broccoli residues fermented with probiotics can be a novel biological feed additive to protect the performance and health of broilers against C. perfringens infection.(AU)

Effect of broccoli residues fermented with probiotics on the growth performance and health status of broilers challenged with Clostridium perfringens

This study aimed at investigating the effects of broccoli residues fermented with probiotics (BF) on the growth performance, immunity, and gut health in broilers challenged with Clostridium perfringens (C. perfringens). A total of 600 broilers (one day old) were randomly allotted into five treatments with six replicates of 20 birds each and were reared until 42 days of age. The treatments included a positive control (PC, fed a basal diet and reared on uncontaminated litter), a negative control (NC, birds reared on litter contaminated with C. perfringens and fed a basal diet), and NC plus BF at 25, 50 or 75 g/kg of diet. The BF contained yeast 3.1 × 10 (7) cfu/g, lactic acid bacteria 9.5 × 10 (6) cfu/g and Bacillus subtilis 3.5 × 10 (6) cfu/g. Birds in the NC group showed lower (p<0.05) feed intake and body weight gain, whereas BF supplementation recovered (p<0.05) the growth performance to the levels of PC group. Dietary BF at 50and 75 g/kg reduced (p<0.05) broiler mortality. Similarly, compared to the NC group, BF increased (p<0.05) immune organ weights and serum immunoglobulins A, G, and M to the levels of PC group. The ileal populations of Escherichia coli and Gram-negative bacteria were decreased (p<0.05) by BF to the levels of PC, and C. perfringens was also decreased (p<0.05) by BF. The serum profiles of mono- and di-amine oxidase were decreased (p<0.05) by BF. BF at 75 g/kg reduced (p<0.05) monoamine oxidase compared with the other BF doses. The results suggest that broccoli residues fermented with probiotics can be a novel biological feed additive to protect the performance and health of broilers against C. perfringens infection.

Active vitamin D3, 1,25-(OH)2D3, protects against macrovasculopathy in a rat model of type 2 diabetes mellitus.

To investigate the protective effect of the active form of vitamin D3, 1,25-(OH)2D3, on macrovasculopathy in rats with type 2 diabetes mellitus (T2DM), 8-week-old male Sprague-Dawley rats were randomly divided into control group, T2DM group, and treatment group. The T2DM model was established after 6 weeks by administering an intraperitoneal injection of streptozotocin (30 mg/kg). 1,25-(OH)2D3 was administered by gavage to rats in the treatment group, and an equal volume of peanut oil was administered to rats in the T2DM group. Fasting plasma glucose (FPG), triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL-C) and low-density lipoprotein (LDL-C) cholesterols were measured in all rats. The morphology of the thoracic aorta was examined, and the expression of tumor necrosis factor alpha (TNF-α), endothelin (ET), endothelial nitric oxide synthase (eNOS), CD54, and CD106 in the thoracic aorta was determined by immunohistochemistry. The expression of FPG, TG, TC, and LDL-C in rats from the T2DM and treatment groups was significantly elevated compared with rats from the control group (P < 0.05). Compared with that in control group, the expression of TNF-α, ET, eNOS, and CD106 was significantly upregulated in the T2DM group and the treatment group, while the expression of CD54 was increased only in the T2DM group (P < 0.05). Moreover, the levels of TNF-α, CD54, and CD106 in rats from the treatment group were lower than those in the T2DM group (P < 0.05). These data suggest that 1,25-(OH)2D3 may protect the macrovessels from injury in T2DM rats by inhibiting the expression of TNF-α, CD54, and CD106.

Meta-analysis of the correlation between the TNF-α308G/A polymorphism and polycystic ovary syndrome.

Previous studies have suggested that the tumor necrosis factor alpha (TNF-α) gene 308G/A polymorphism may be associated with polycystic ovary syndrome (PCOS) risk. However, this relationship is controversial. The present meta-analysis aimed to evaluate the correlation between the TNF-α308G/A polymorphism and susceptibility to PCOS. A systematic electronic search of PubMed and Embase databases was conducted using specific inclusion criteria. Summary odds ratios (ORs) and 95% confidence intervals (95%CIs) were calculated, and all statistical analyses were performed using STATA 12.0. The results of our meta-analysis showed no significant association between the TNF-α308G/A polymorphism and PCOS risk (AA vs GG: OR = 0.80, 95%CI = 0.31-2.08; AG vs GG: OR = 1.03, 95%CI = 0.59-1.81; dominant model: OR = 1.02, 95%CI = 0.60-1.71; recessive model: OR = 0.87, 95%CI = 0.35-2.16). Based on the statistical data, our meta-analysis indicates that the TNF-α308G/A sequence variation may be not related to PCOS susceptibility. Further large and well-designed studies are needed to confirm this conclusion.

Association of IL-18 polymorphisms with rheumatoid arthritis: a meta-analysis.

Interleukin-18 (IL-18), an important proinflammatory cytokine, has been reported to play a potential pathological role in rheumatoid arthritis (RA). Results from previous studies on the association between IL-18 polymorphisms and RA are conflicting. To clarify this, an updated meta-analysis of all available studies on IL-18 polymorphisms and RA was conducted. Eligible articles were identified by searching databases, including PubMed, Ovid, Cochrane Library, EMBASE, and China Knowledge Resource Integrated Database, for the period up to May 1, 2015. The pooled odds ratios (ORs) with 95% confidence intervals (95%CIs) were used to assess the strength of association in the homozygote, heterozygote, dominant, recessive, and additive models. The software STATA (Version 13.0) was used for statistical analysis. Finally, 14 articles were included in the present meta-analysis. The IL-18 -607C/A polymorphism showed pooled ORs and 95%CIs for the homozygote model (AA vs CC: OR = 0.598; 95%CI = 0.395-0.907), and the association between the IL-18 -137G/C polymorphism and RA showed pooled ORs and 95%CIs for the homozygote (CC vs GG: OR = 0.699; 95%CI = 0.364-1.342) and heterozygote (CG vs GG: OR = 0.924; 95%CI = 0.803-1.064) models. In summary, the current meta-analysis, which was based on the most current studies, showed that the -607A/C, -920C/T, and -105A/C polymorphisms in IL-18 were significantly associated with increased RA risk. However, the -137C/G polymorphism was not associated with RA risk under any genetic model. More evidence is needed to support or deny such a conclusion.

Detection of Trypanosoma cruzi DNA in blood by PCR is associated with Chagas cardiomyopathy and disease severity.

BACKGROUND: The significance of detection of Trypanosoma cruzi DNA in blood of antibody-positive patients for risk of development of Chagas heart disease is not well established. The objective of this study was to compare detection of T. cruzi DNA with known clinical and laboratory markers of Chagas cardiomyopathy (CC) severity. METHODS: This is a case-control study nested within a retrospective cohort developed in Brazil to understand the natural history of Chagas disease. The study enrolled 499 T. cruzi seropositive blood donors (SP-BD) and 488 frequency matched seronegative control donors (SN-BD) who had donated between 1996 and 2002, and 101 patients with clinically diagnosed CC. In 2008-2010 all enrolled subjects underwent a health questionnaire, medical examination, electrocardiograms and echocardiograms and polymerase chain reaction (PCR) analyses. A blinded panel of three cardiologists adjudicated the outcome of CC. Trypanosoma cruzi kinetoplast minicircle sequences were amplified by real-time PCR using an assay with a sensitivity of one parasite per 20 mL of blood. All testing was performed on coded samples. RESULTS: Rates of PCR detection of T. cruzi DNA were significantly (P = 0.003) higher in CC patients and SP-BD diagnosed with CC (79/105 [75.2 %]) compared with SP-BD without CC (143/279 [51.3%]). The presence of parasitaemia was significantly associated with known markers of disease progression such as QRS and QT interval duration, lower left ventricular ejection fraction, higher left ventricular index mass, and elevated troponin and NTpro-BNP levels. CONCLUSION: Trypanosoma cruzi PCR positivity is associated with presence and severity of cardiomyopathy, suggesting a direct role of parasite persistence in disease pathogenesis.

High expression of HMOX1 in blue-shelled chickens is associated with a TG haplotype

HMOX1 is an important gene in biosynthesis of the eggshell pigment of blue eggs. Previous studies found that HMOX1 is highly expressed in the shell gland of hens laying blue eggs (BlueH) compared with hens laying brown eggs (BrownH); however, the reasons for the differential expression are unclear. In this study five single nucleotide polymorphism (SNP) in HMOX1 were genotyped in 111 BlueH and 115 BrownH. The association of haplotypes of these SNP with the blue egg phenotype was tested. Haplotype-specific expression of HMOX1 was detected in the shell gland. The interaction of sequence variants and transcription factors was analyzed using electrophoretic mobility shift assay (EMSA). A TG haplotype covering upstream 1.4 kb region of HMOX1 was significantly associated with blue eggs (p 0.05). Furthermore, the birds (n=12) with the haplotype expressed 3.8 fold more transcripts than those (n=12) without the haplotype (p 0.05). After re-sequencing a 2.2 kb region harboring the TG haplotype, a total of 26 SNP were found, of which a SNP was predicted to create a binding site of Nrf2, a transcription factor initiating HMOX1 expression. However, subsequent EMSA failed to confirm the Nrf2-DNA interaction. Taken together, the data suggested that the TG haplotype is not directly involved in regulation of HMOX1 expression; a regulatory mutation located near the haplotype and linked with the haplotype may exist and be responsible for the differential expression of HMOX1.

High expression of HMOX1 in blue-shelled chickens is associated with a TG haplotype

HMOX1 is an important gene in biosynthesis of the eggshell pigment of blue eggs. Previous studies found that HMOX1 is highly expressed in the shell gland of hens laying blue eggs (BlueH) compared with hens laying brown eggs (BrownH); however, the reasons for the differential expression are unclear. In this study five single nucleotide polymorphism (SNP) in HMOX1 were genotyped in 111 BlueH and 115 BrownH. The association of haplotypes of these SNP with the blue egg phenotype was tested. Haplotype-specific expression of HMOX1 was detected in the shell gland. The interaction of sequence variants and transcription factors was analyzed using electrophoretic mobility shift assay (EMSA). A TG haplotype covering upstream 1.4 kb region of HMOX1 was significantly associated with blue eggs (p 0.05). Furthermore, the birds (n=12) with the haplotype expressed 3.8 fold more transcripts than those (n=12) without the haplotype (p 0.05). After re-sequencing a 2.2 kb region harboring the TG haplotype, a total of 26 SNP were found, of which a SNP was predicted to create a binding site of Nrf2, a transcription factor initiating HMOX1 expression. However, subsequent EMSA failed to confirm the Nrf2-DNA interaction. Taken together, the data suggested that the TG haplotype is not directly involved in regulation of HMOX1 expression; a regulatory mutation located near the haplotype and linked with the haplotype may exist and be responsible for the differential expression of HMOX1.(AU)

Expression profiling of CEACAM6 associated with the tumorigenesis and progression in gastric adenocarcinoma.

Carcinoembryonic antigen-related cellular adhesion molecule 6 (CEACAM6) is a member of the immunoglobulin superfamily and has been recently reported to affect the neoplastic, metastatic, and invasive ability of malignant cells by regulating intracellular signaling pathways during tumorigenesis and progression. We investigated the expression and amplification of CEACAM6 in relation to the clinicopathological and biological significance of gastric adenocarcinoma. Expression of CEACAM6 mRNA in 75 primary gastric adenocarcinom and 20 adjacent tissues compared to normal gastric mucosas were explored using real-time quantitative-polymerase chain reaction. Immunohistochemical assays were conducted to evaluate the expression and tissue distribution of CEACAM6 protein. Overexpression of CEACAM6 mRNA in both gastric adenocarcinoma (2.513 ± 0.869) and adjacent tissues (1.171 ± 0.428) was significantly higher than the relative expressions in non-neoplastic specimens (0.594 ± 0.513) (P < 0.01). CEACAM6 protein was present in 52 (69.33%) gastric adenocarcinomas, but not in normal gastric tissues. Adenocarcinomas with elevated CEACAM6 expression were significantly associated with lymph node metastases and advanced stages. There were no relationships between CEACAM6 expression and tumor size, histological differentiation, or different subtypes, respectively. Moreover, higher expression of CEACAM6 was found to be correlated with short postoperative survival time of patients with gastric cancer. Amplification and upregulation of CEACAM6 expression was observed in human gastric adenocarcinomas, which may be correlated with the generation or transformation of malignant cells, tumor aggressive progression, and clinical outcome. CEACAM6 may be a valuable biomarker screening for gastric tumor and novel predictor for patients in advanced stages of gastric cancer.

A case-control study indicates that the TRIB1 gene is associated with pancreatic cancer.

Pancreatic cancer is a malignant neoplasm originating from transformed cells arising in tissues that form the pancreas. To investigate whether the tribbles homolog 1 (Drosophila) gene (TRIB1) is associated with pancreatic cancer in the Chinese Han population, we conducted this case-control study and genotyped 3 single nucleotide polymorphisms (rs2980879, rs2980874, and rs2235108) of the TRIB1 gene in 182 patients and 359 normal controls of Chinese Han origin and analyzed their association. The results showed that the rs2980879 polymorphism was associated with pancreatic cancer [allele: P = 0.023434, genotype: P = 0.03005; odds ratio (OR) and 95% confidence interval (CI) = 0.727788 (0.552664-0.958404)], whereas the rs2980874 polymorphism had no association with pancreatic cancer [allele: P = 0.749885, genotype: P = 0.699533; OR and 95%CI = 1.041981 (0.809196-1.341734)], and the rs2235108 polymorphism was not associated with the disease [allele: P = 0.629475, genotype: P = 0.547534, OR and 95%CI = 1.128290 (0.690829-1.842770)]. Haplotype analyses and linkage disequilibrium tests were also conducted, and the results showed that these 3 loci are not in the same block. In conclusion, our study indicated that the TRIB1 gene is associated with pancreatic cancer. More studies with larger samples are needed in order to support this finding.

Gene expression profiling of Duchenne muscular dystrophy reveals characteristics along disease progression.

Duchenne muscular dystrophy (DMD) is the most common form of muscular dystrophy with no cure currently available. In this study, using two microarray data sets obtained from the Gene Expression Omnibus database, we conducted a dysfunctional pathway-enrichment analysis and investigated deregulated genes that are specific to different phases of the disease in order to determine pathogenic characteristics in the progression of DMD. We identified 41 and 33 dysfunctional pathways that were enriched with differentially expressed genes in presymptomatic patients and in symptomatic patients, respectively. Over 70% of pathways were shared between both phases and many of them involved the inflammatory process, suggesting that inflammatory cascades were induced soon after the birth of the patients. Further investigation showed that presymptomatic patients performed better with respect to muscle regeneration and cardiac muscle calcium homeostasis maintenance. Neuronal nitric oxide synthase, dihydropyridine receptors, sarcoplasmic/endoplasmic reticulum calcium ATPase, and phospholamban may serve as potential targets for further molecular diagnostic tests. Our results may provide a better understanding for the treatment of DMD.

Edaravone protects endotoxin-induced liver injury by inhibiting apoptosis and reducing proinflammatory cytokines

Studies have shown that edaravone may prevent liver injury. This study aimed to investigate the effects of edaravone on the liver injury induced by D-galactosamine (GalN) and lipopolysaccharide (LPS) in female BALB/c mice. Edaravone was injected into mice 30 min before and 4 h after GalN/LPS injection. The survival rate was determined within the first 24 h. Animals were killed 8 h after GalN/LPS injection, and liver injury was biochemically and histologically assessed. Hepatocyte apoptosis was measured by TUNEL staining; proinflammatory cytokines [tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)] in the liver were assayed by ELISA; expression of caspase-8 and caspase-3 proteins was detected by Western blot assay; and caspase-3 activity was also determined. Results showed that GalN/LPS induced marked elevations in serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Edaravone significantly inhibited elevation of serum AST and ALT, accompanied by an improvement in histological findings. Edaravone lowered the levels of TNF-α and IL-6 and reduced the number of TUNEL-positive cells. In addition, 24 h after edaravone treatment, caspase-3 activity and mortality were reduced. Edaravone may effectively ameliorate GalN/LPS-induced liver injury in mice by reducing proinflammatory cytokines and inhibiting apoptosis.

Edaravone protects endotoxin-induced liver injury by inhibiting apoptosis and reducing proinflammatory cytokines.

Studies have shown that edaravone may prevent liver injury. This study aimed to investigate the effects of edaravone on the liver injury induced by D-galactosamine (GalN) and lipopolysaccharide (LPS) in female BALB/c mice. Edaravone was injected into mice 30 min before and 4 h after GalN/LPS injection. The survival rate was determined within the first 24 h. Animals were killed 8 h after GalN/LPS injection, and liver injury was biochemically and histologically assessed. Hepatocyte apoptosis was measured by TUNEL staining; proinflammatory cytokines [tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)] in the liver were assayed by ELISA; expression of caspase-8 and caspase-3 proteins was detected by Western blot assay; and caspase-3 activity was also determined. Results showed that GalN/LPS induced marked elevations in serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Edaravone significantly inhibited elevation of serum AST and ALT, accompanied by an improvement in histological findings. Edaravone lowered the levels of TNF-α and IL-6 and reduced the number of TUNEL-positive cells. In addition, 24 h after edaravone treatment, caspase-3 activity and mortality were reduced. Edaravone may effectively ameliorate GalN/LPS-induced liver injury in mice by reducing proinflammatory cytokines and inhibiting apoptosis.

Structural analysis of a 4414-bp element in Drosophila melanogaster.

We cloned a 4414-bp element from a mutant of Drosophila melanogaster. Its insertion site was 18,929,626 bp. Analysis of the nucleotide and amino acid sequences demonstrated that the element is homologous to Pifo_I, first obtained from D. yabuka, which belongs to the gypsy/Ty3 subfamily. We also obtained a 3754-bp length element from a wild-type fly by PCR, with a pair of primers designed from the conserved region of the 4414-bp length element. The two elements included a pair of long terminal repeats and part of the GAG and ENV proteins, but the POL protein was completely lost. This element is found in the subgenus of D. melanogaster, but it is a degenerate type of Pifo_I and is not infective. Also, a 714-bp region structured in 5.0 tandem repeats of 143 bp each was found in the 5'UTR of the degenerate element; these could interact with transcription factor CF2. Phylogenetic analysis and alignment of amino acids indicated that the Pifo_I element was closer to the ZAM retrotransposon, which gave us some clues to their functional similarity. Based on these data, we propose that there is a relationship between the degenerate element and the mutant phenotype, which would provide a foundation for further research.

In vitro protein release and degradation of poly-dl-lactide-poly(ethylene glycol) microspheres with entrapped human serum albumin: quantitative evaluation of the factors involved in protein release phases.

PURPOSE: To quantitatively evaluate the correlations between the amount of initial burst release and the surface-associated protein, and between the onset time for the second burst release and the matrix polymer degradation. METHODS: Human serum albumin (HSA) was microencapsulated in polylactide (PLA) and poly-dl-lactide-poly(ethylene glycol) (PELA) with PEG contents of 5, 10, 20, and 30%, respectively, using the solvent extraction procedure based on formation of double emulsion w/o/w. Microspheres with similar particle size (1.7-2.0 microm), similar protein entrapment (2.1-2.8%) but different surface-associated proteins (9.3-53.6%) were used to evaluate the in vitro matrix degradation and protein release profiles. Degradation was characterized by studying the intrinsic viscosity decrease, medium pH change, and weight loss of the microspheres. RESULTS: The matrix degradation and protein release profiles were highly dependent on the polymer composition of the microspheres. Faster decreases in the intrinsic viscosity of recovered matrix polymer, the microspheres weight, and the pH of degradation medium, and earlier onsets for the break in intrinsic viscosity reduction and the mass loss were detected for PELA microspheres with higher PEG content. The hydration and swelling of microspheres matrix contributed greatly to the degradation of matrix polymer. The HSA release showed triphasic profile and involved two mechanisms for all the microsphere samples. Smaller amount of initial burst release, larger gradual release rate, and earlier onset for the second burst release were observed for HSA from matrix polymer with higher PEG content. The extent of the initial burst release was quantitatively related with the surface-associated protein. The second burst release of HSA was observed to occur within 1 week after the onset for mass loss, which was also the break in the intrinsic viscosity reduction rate. CONCLUSION: Protein release profiles could be rationalized by optimizing the matrix polymer degradation and microsphere characteristics.