Hen- Wei

Nodal promotes growth and invasion in human gliomas.

Uncontrolled growth and diffused invasion are major causes of mortality in patients with malignant gliomas. Nodal has been shown to have a central role in the tumorigenic signaling pathways of malignant melanoma. In this study, we show that grade IV human glioma cell lines expressed different levels of Nodal, paralleled to the potential for cell invasiveness. Treatment of glioma cell lines with recombinant Nodal (rNodal) increased matrix metalloproteinase 2 (MMP-2) secretion and cell invasiveness. The ectopic expression of Nodal in GBM glioma cells that expressed Nodal at low level resulted in increased MMP-2 secretion, enhanced cell invasiveness, raised cell proliferation rates in vitro, increased tumor growth in vivo, and was associated with poor survival in a mice xenograft model. In contrast, the knockdown of Nodal expression in U87MG glioma cells with high Nodal expression level had reduced MMP-2 secretion, less cell invasiveness, lower tumor growth in vivo and longer lifespan in mice with U87MG/shNodal cell xenografts. In addition, Nodal knockdown promoted the reversion of malignant glioma cells toward a differentiated astrocytic phenotype. Furthermore, our data support the notion that Nodal may regulate glioma progression through the induction of the leukemia inhibitory factor (LIF) and Cripto-1 through activated Smad.

Dexamethasone reduced invasiveness of human malignant glioblastoma cells through a MAPK phosphatase-1 (MKP-1) dependent mechanism

Dexamethasone has been shown to inhibit tumor invasiveness. In the present study, the effects of dexamethasone on matrix metalloproteinases-2 (MMP-2) secretion, cell invasiveness, and intravasation in human U87MG glioma cells were examined. Dexamethasone decreased MMP-2 secretion and cell invasiveness in human glioma cells. Incubation of cells with dexamethasone increased mitogen activated protein kinase phosphatase-1 (MKP-1) expression. Ectopic expression of MKP-1 decreased cell invasiveness in vitro and intravasation in vivo. Because expression of inducible nitric oxide synthase (iNOS) has been implicated in the progression of malignant gliomas, we next investigated the possible roles of NO(-) in MMP-2 secretion and cell invasiveness in human U87MG glioma cells. Treatment of glioma cells with nitric oxide donor, sodium nitroprusside (SNP), increased MMP-2 secretion and the capacity of cell invasion in U87MG cells. Addition of dexamethasone or ectopic expression of wild-type MKP-1 suppressed the SNP-stimulated MMP-2 activation and glioma cell invasiveness in U87MG cells. Taken together, these results suggest that dexamethasone may suppress MMP-2 secretion and cell invasion through MKP-1 induction in human glioma cells.

Antimicrobial resistance of Escherichia coli isolates from canine urinary tract infections.

This study determined the antimicrobial resistance profiles of Escherichia coli isolates from dogs with a presumptive diagnosis of urinary tract infection (UTI). Urine samples from 201 dogs with UTI diagnosed through clinical examination and urinalysis were processed for isolation of Escherichia coli. Colonies from pure cultures were identified by biochemical reactions (n=114) and were tested for susceptibility to 18 antimicrobials. The two most frequent antimicrobials showing resistance in Urinary E. coli isolates were oxytetracycline and ampicillin. Among the resistant isolates, 17 resistance patterns were observed, with 12 patterns involving multidrug resistance (MDR). Of the 69 tetracycline-resistant E. coli isolates, tet(B) was the predominant resistance determinant and was detected in 50.9% of the isolates, whereas the remaining 25.5% isolates carried the tet(A) determinant. Most ampicillin and/or amoxicillin-resistant E. coli isolates carried blaTEM-1 genes. Class 1 integrons were prevalent (28.9%) and contained previously described gene cassettes that are implicated primarily in resistance to aminoglycosides and trimethoprim (dfrA1, dfrA17-aadA5). Of the 44 quinolone-resistant E. coli isolates, 38 were resistant to nalidixic acid, and 6 were resistant to nalidixic acid, ciprofloxacin and enrofloxacin. Chromosomal point mutations were found in the GyrA (Ser83Leu) and ParC (Ser80Ile) genes. Furthermore, the aminoglycoside resistance gene aacC2, the chloramphenicol resistant gene cmlA and the florfenicol resistant gene floR were also identified. This study revealed an alarming rate of antimicrobial resistance among E. coli isolates from dogs with UTIs.

Rosiglitazone reduces cell invasiveness by inducing MKP-1 in human U87MG glioma cells.

We sought to investigate the molecular mechanisms by which rosiglitazone (RGZ) inhibits cell invasion in human glioma cells. In this study, we found that RGZ attenuated MMP-2 protein levels, MMP-2 gelatinolytic activity, and cell invasiveness through a PPAR-gamma independent pathway. RGZ increased mitogen activated protein kinase phosphatase-1 (MKP-1) expression. The addition of triptolide (a diterpenoid triepoxide, which blocked MKP-1 induction) abolished the inhibitory effects by RGZ. Furthermore, we demonstrated that the knock down of MKP-1 by MKP-1 specific small interference RNA reversed the reduction of MMP-2 secretion, and of cell invasiveness by RGZ. In contrast, the stable expression of MKP-1 in glioma cell lines decreased MMP-2 activity and cell invasiveness. These results suggest that RGZ may mediate the inhibitory effects through MKP-1 induction. Thus, MKP-1 could be a potential target in glioma therapy.

Generation and Utilization of P450 Cholesterol Side‐Chain Cleavage Enzyme and 3β‐Hydroxysteroid Dehydrogenase Antibodies for Universal Detection

Abstract The biosynthesis of steroids from steroidogenic cells are catalyzed by the two major enzymes, P450 side‐chain cleavage enzyme (P450scc) and 3β‐hydroxysteroid dehydrogenase (3β‐HSD). This article describes the construction of two novel polyclonal antibodies against conserved recombinant protein and the validation of these antibodies on fixed tissue sections of bovine corpus luteum. The polyclonal antibodies were used successfully in Western blots and specifically reacted with P450scc and 3β‐HSD protein in bovine luteal cell extracts. Thus, P450scc and 3β‐HSD are two specific polyclonal antibodies that are integral products in the investigation of the biological function and regulatory mechanism involved in steroidogenesis.

Dietary deficiencies of single amino acids: Whole-body amino acid composition of adult rats

Abstract To ascertain whether chronic amino acid deficiency alters the amino acid composition of the body, 44 adult female rats were randomly allocated to one of 11 treatments which included one control group, ingesting an adequate diet with balanced protein, and ten deficient groups in which one group received protein-deficient diets and the other groups consumed diets each deficient in a single essential amino acid. The degree of deficiency was adjusted to achieve a gradual decline in body weight to 85% of the initial weight and was then adjusted so that this weight was maintained until the end of the experiment at 93 days, when the rats were killed. Deficient rats had lower absolute weights of liver, gastrointestinal tract and muscle than animals given the adequate diet but greater relative weights (% of body weight) of heart and kidneys. There were no significant differences amongst groups in percentages of lipid, nitrogen, protein plus lipid or dry matter. Chronic marginal amino acid deficiencies did not selectively alter amino acid composition.

Optimization of the thermal conditions for processing hatchery waste eggs as meal for feed

The purpose of this study was to optimize the thermal conditions for processing hatchery waste eggs (HWE) into rich feedstuff with lower electricity consumption by using response surface methodology. In the study, the effects of processing temperature and time on HWE meal (HWEM) quality and production were evaluated. As the results indicate, optimization was obtained when the processing lasted for 23 h at the fixed temperature of 65°C, resulting in higher protein digestibility in vitro (89.6%) and DM (88.5%) content of HWEM with lower electricity consumption (82.4 kWh/60 kg of HWE). No significant differences existed between the quality values predicted by mathematical formulae and those obtained through practical analyses in DM (87 vs. 88.5%), CP (39.2 vs. 38.3%), protein digestibility in vitro (90.7 vs. 89.6%), and electricity consumed (80.8 vs. 82.4 kWh/60 kg of HWE). Furthermore, the product derived from the optimized processing conditions had better biosecurity; Salmonella spp. were not found and Escherichia coli levels were substantially reduced (from 10(7) to 10(4) cfu/g). In summary, HWEM of superior quality can be produced when the processing conditions optimized in the current research are utilized.

Estimating the requirement of dietary crude protein for growing blue-breasted quail ( Excalfactoria chinensis )

Two experiments were conducted to investigate the requirement for dietary crude protein (CP) in growing blue-breasted quail (BBQ). In Experiment 1, 300 1-day-old quails were randomly assigned to 10 groups according to a 2×5 factorial arrangement of treatments with two metabolisable energy (ME) levels (12.13 and 13.39 MJ/kg) and five CP concentrations (160, 190, 220, 250 and 280 g/kg) for 8 weeks. In Experiment 2, 300 1-day-old quails were subjected to a different factorial arrangement of treatments with two ME levels (11.51 and 12.13 MJ/kg) and five CP concentrations (210, 220, 230, 240 and 250 g/kg) for 28 days. Experiment 1 revealed that an interaction existed in weight gain between ME and CP levels in weeks 1 to 4. In both ME groups, quails receiving CP of 160 g/kg showed the least weight gains (P<0.05). No differences (P>0.05) existed in weight gain between the ME groups in which quails ingested CP of 250 and 280 g/kg, whereas quails consuming CP of 220 g/kg with an ME of 13.39 MJ/kg had smaller weight gain than did those ingesting higher CP concentrations (P<0.05). Of main effects for weeks 1-4, quails treated with an ME of 12.13 MJ/kg consumed more feed than did those receiving another ME level, whereas quails in both ME treatments showed similar feed efficiencies. For weeks 5 to 8, no difference (P>0.05) in weight gain, feed intake and feed efficiency was seen regardless of ME levels, and no interaction existed between ME and CP levels. In Experiment 2, the best weight gain and feed efficiency were achieved when the dietary CP concentration was more than 210 g/kg, and quails treated with 11.51 MJ/kg showed better weight gain and feed efficiency (P<0.05) than did those that received 12.13 MJ/kg. Furthermore, the weight gains and protein intakes on the basis of per MJ from the two experiments were pooled together to estimate the protein intake necessary for the best growth performance by two mathematic models; they were then converted to dietary CP concentrations of 204 (minimum) and 233 g/kg (maximum) when ME was 11.51 MJ/kg. In conclusion, BBQ will achieve good growth performance with dietary CP of more than 204 g/kg on the basis of an ME of 11.51 MJ/kg in weeks 1 to 4.

Apportioning protein requirements for maintenance v. growth for blue-breasted quail (Excalfactoria chinensis) from 7 to 21 days of age.

The aim of this study was to investigate protein requirements for the maintenance and growth of blue-breasted quail (Excalfactoria chinensis) from 7 to 21 days of age. A total of 180 quails, 7 days old, were randomly assigned to 36 cages and for 2 weeks were fed diets with a metabolisable energy concentration of 12.13 MJ/kg and a dietary CP concentration of 125, 150, 175, 200, 225 or 250 g/kg. The average BW per cage and the feed intake per cage were recorded daily. The results showed that quails fed 125 g/kg CP could not maintain their BW and had negative feed efficiency. There were linear and quadratic relationships between CP level and response criteria, including BW, weight gain, feed intake, feed efficiency, final body nitrogen mass and body nitrogen accretion (P<0.05). The dietary CP requirements, as calculated using a one-slope quadratic broken-line model, were 211 and 202 g/kg according to weight gain and feed efficiency, respectively. The regression equations, on the basis of metabolic BW, of daily weight gain on daily protein intake according to the model were Y=0.137-2.128(0.113-X) if X<0.113 and Y=0.137 if X>or=0.113 (R2=0.96, P<0.001), which meant that the protein requirement for maintenance was 0.049 times the metabolic BW and that to gain 1 g weight quails needed to ingest an extra 0.47 g protein after the maintenance requirement was satisfied. The regression equations, on the basis of metabolic BW, of daily body nitrogen accretion on daily protein intake according to the model were Y=5.667-76.700(0.119-X) if X<0.119 and Y=5.667 if X>or=0.119 (R2=0.95, P<0.001), which meant that quails had to receive an amount of protein equal to their metabolic BW multiplied by 0.045 to satisfy the requirement for maintenance and then ingest an extra 13 g protein to accrete 1 g body nitrogen. In conclusion, growth or protein accretion rates should be regulated according to dietary CP for specific experimental purposes via apportioning protein requirements for maintenance v. growth.