[Factors influencing bilirubin elevation and its correlation with UGT1A1 gene polymorphism in the early postoperative period of transjugular intrahepatic portosystemic shunt].

Objective: To investigate the factors influencing total bilirubin elevation and its correlation with UGT1A1 gene polymorphism in the early postoperative period of transjugular intrahepatic portosystemic shunt (TIPS). Methods: 104 cases with portal hypertension and esophageal variceal hemorrhage (EVB) treated with elective TIPS treatment were selected as the study subjects and were divided into a bilirubin-elevated group and a normal bilirubin group according to the total bilirubin elevation level during the early postoperative period. Univariate analysis and logistic regression were used to analyze the factors influencing total bilirubin elevation in the early postoperative period. PCR amplification and first-generation sequencing technology were used to detect the polymorphic loci of the UGT1A1 gene promoter TATA box, enhancer c.-3279 T > G, c.211G > A, and c.686C > A. Logistic regression was used to analyze the correlation of four locus alleles and genotypes with elevated total bilirubin in the early postoperative period. Results: Among the 104 cases, 47 patients were in the bilirubin elevated group, including 35 males (74.5%) and 12 females (25.5%), aged (50.72 ± 12.56) years. There were 57 cases in the normal bilirubin group, including 42 males (73.7%) and 15 females (26.3%), aged (51.63 ± 11.10) years. There was no statistically significant difference in age (t = -0.391, P = 0.697) and gender (χ(2) = 0.008, P = 0.928) between the two groups of patients. Univariate analysis revealed that preoperative alanine transaminase (ALT) level (χ(2) = 5.954, P = 0.015), total bilirubin level (χ(2) = 16.638, P < 0.001), MELD score (χ(2) = 10.054, P = 0.018), Child-Pugh score (χ(2) = 6.844, P = 0.022), and postoperative portal vein branch development (χ(2) = 6.738, P = 0.034) were statistically significantly different between the two groups. Logistic regression analysis showed that preoperative ALT level, total bilirubin level, and portal vein branch development after TIPS were correlated with the elevated total bilirubin in the early postoperative period. The polymorphism of the c.211G > A locus of the UGT1A1 gene correlation had elevated total bilirubin in the early postoperative period of TIPS. The risk of elevated total bilirubin was increased in the population carrying allele A (P = 0.001, OR = 4.049) in the early postoperative period. Allelic polymorphisms in the TATA box promoter region and enhancer c.-3279 T > G and c.686C > A had no statistically significant difference between the bilirubin-elevated group and the normal bilirubin group. Conclusion: The preoperative ALT level, total bilirubin level, and portal vein branch development are correlated with the elevated total bilirubin in early postoperative patients. The polymorphisms of the UGT1A1 gene and enhancer c.211G > A are correlated with the occurrence of elevated total bilirubin in the early postoperative period of TIPS. Allele A carrier may have a higher risk of elevated total bilirubin in the early postoperative period.

2-Nitroimidazole based fluorescent probes for nitroreductase; monitoring reductive stress in cellulo.

Two 2-nitroimidazole-1,8-naphthalimide conjugates, 1 and 2, have been synthesised as fluorescence probes for the detection of reductive stress in HeLa cells. The 4-substituted derivative 1 was shown to act as a highly sensitive and selective substrate for nitroreductase where it exhibited a clear blue to green ratiometric fluorescence response visible to the naked eye. Moreover, biological studies demonstrated 1 could be activated in cellulo where the impact of reductive stress was easily monitored using confocal microscopy and flow cytommetry.

Identification of lactic acid bacteria in traditional fermented yak milk and evaluation of their application in fermented milk products.

In this study, 53 strains of lactic acid bacteria (LAB) isolated from Xueo, a traditional fermented yak milk in the western Sichuan Plateau of China, were identified and their use in fermented milk was evaluated. All gram-positive and catalase-negative strains were divided into 6 groups at the level of 87% similarity using amplified ribosomal DNA restriction analysis. These groups were identified as 6 species using 16S rDNA sequence analysis and atpA gene analysis. The dominant LAB strains in Xueo were Enterococcus durans, Lactobacillus fermentum, and Lactobacillus paracasei, accounting for 45.3, 22.6, and 17.0% of isolates, respectively. Milk fermented with most of the representative strains was high in quality, exhibiting relatively high viscosity, moderate acidity, good sensory quality, and high counts of viable LAB. Fermented milk of E. durans SCA16 and L. fermentum SCA52 achieved the highest scores for overall sensory quality. Most strains displayed antimicrobial activity against at least 1 of 9 spoilage microorganisms. Lactic acid was the main factor inhibiting the growth of spoilage bacteria, and H(2)O(2) was also inhibitory to some extent. Excluding the influence of acid and H(2)O(2), strains SCA52 (L. fermentum) and SCA7 (Lactobacillus plantarum) were antagonistic against some of the indicators, suggesting that the 2 strains may produce a bacteriocin-like substance. Therefore, the development of superior strains isolated from Xueo to ferment milk with similar flavor and texture to Xueo is expected.

Effect of increased levels of adiponectin by administration of the adeno vector rAAV2/1-Acrp30 on glucose, lipid metabolism and ultrastructure of the aorta in Goto-Kakizaki rats with arteriosclerosis.

BACKGROUND: We used recombinant adeno-associated virus vector of adiponectin (AAV2/1-Acrp30) to study the effects of increased levels of adioponectin (by the administration of rAAV2/1-Acrp30) on arteriosclerosis, glucose and lipid metabolism in Goto-Kakizaki (GK) rats with arteriosclerosis. METHODS: Thirty GK rats with arteriosclerosis were divided into 3 equal groups: control group 1, control group 2 and the rAAV2/1-Acrp30-administered group. Saline, virus vector or rAAV2/1-Acrp30 (10 12 ng/ml) vector genomes administered to the rats in the corresponding group by intramuscular injection to the posterior limb by single administration, respectively. After 8 weeks, fasting blood glucose, 2-hour postprandial blood glucose, glycosylated haemoglobin, serum insulin, serum total cholesterol, triglycerides, high-density lipoprotein and low-density lipoprotein were measured in each group, and the ultrastructure of the aorta was seen by light and electron microscopy. RESULTS: Compared with control groups 1 and 2, in the rAAV2/1-Acrp30 group, there was a decrease in urine volume, fasting blood glucose, 2-hour postprandial blood glucose, glycosylated haemoglobin, serum total cholesterol, triglycerides and low-density lipoprotein, and an increase in body weight and high-density lipoprotein (p< 0.05), while the level of serum insulin was not changed (p>0.05). Ultrastructure studies of the aorta showed that aortosclerosis in the rAAV2/1-Acrp30-administered group was less, and fewer lipid droplet vacuoles were seen in the vascular endothelial cytoplasm. Also various cell organelles and internal elastic lamina were seen, and there was no formation of lipid droplet and foam cells in the cytoplasm of the media of the smooth muscle. CONCLUSION: Adiponectin could improve blood glucose and lipid parameters and decrease atherosclerosis in the aorta of GK rats.

The study of the tumor stem cell properties of CD133+CD44+ cells in the human lung adenocarcinoma cell line A549.

We studied the tumor stem cell properties of the CD133+CD44+ subpopulation in the human lung adenocarcinoma cell line A549. A549 cells were classified into subpopulations based on differential expression patterns for CD133 and CD44. Cells from different subpopulations were cultured and subcutaneously injected into 32 nude mice. Our results as following, (1) The majority of A549 cells died, whereas only about 4.11% of cells divided and proliferated to form cell clones. (2) The expression of CD133 and CD44 in proliferative cancer cells was statistically significantly different from that in normal A549 cells (p < 0.001). (3) Cell proliferation in group A (CD133+CD44+) was the fastest among all groups. Cell proliferation in A549 cells was slower than in group A but faster than in groups B (CD133-CD44-), C (CD133-CD44+), and D (CD133+CD44-). (4) The tumorigenic capacity in cells from group A was significantly higher than that in cells from groups B (p<0.001), C (p<0.001) and D (p<0.04). In conclusion, CD133+CD44+ cells in the adenocarcinoma cell line A549 have expressive significant cancer stem cell properties with continuous proliferative capacity and differentiation potential.

Effects of Achyranthes bidentata polysaccharides on performance, immunity, antioxidant capacity, and meat quality in Pekin ducks.

This study was conducted to evaluate the effects of Achyranthes bidentata polysaccharide (ABP) on growth performance, antioxidant capacity, immune function, relative organ weight, ileal microflora, and meat quality in Pekin ducks. A total of 1,200 female 1-day-old Pekin ducklings (51.2 ± 0.2 g) were blocked based on body weight (BW) and randomly allocated into 3 treatments with 10 replicates of 40 birds each. The experiment lasted for 6 wk, and dietary treatments included corn-soybean meal-based diet supplemented with 0, 0.02, and 0.04% ABP. The supplementation of ABP increased (P < 0.05) body weight gain (BWG) and final BW linearly during day 22 to 42 and day 1 to 42, respectively, but decreased (P < 0.05) feed-to-gain ratio (F/G) linearly during day 22 to 42 and day 1 to 42. The inclusion of ABP increased (P < 0.05) serum superoxide dismutase, glutathione peroxidase, total antioxidative capacity, catalase, complement3, complement4, immunoglobin A, immunoglobin G, interleukin-2, interferon-γ, and tumor necrosis factor-α linearly. The relative weight of breast meat was increased (P < 0.05) linearly, but the relative weight of abdominal fat was decreased (P < 0.05) linearly with the increasing dietary ABP supplementation. The supplementation of ABP increased (P < 0.05) ileal Lactobacilli counts linearly, whereas decreased (P < 0.05) Escherichia coli counts linearly. Taken together, the inclusion of ABP promoted BWG and final BW during day 22 to 42 and the entire experiment, decreased F/G during day 22 to 42 and day 1 to 42, and partially improved antioxidant activities, immunity, and gut microflora in Pekin ducks.

Effects of grape seed extract on performance, immunity, antioxidant capacity, and meat quality in Pekin ducks.

This study was conducted to evaluate the effects of grape seed extract (GSE) on growth performance, immunity, antioxidant capacity, relative organ weight, jejunum morphology, ileal microflora, and meat quality in Pekin ducks. A total of 1,500 female 1-day-old Pekin ducklings (52.0 ± 0.2 g) were blocked based on body weight (BW) and randomly allocated into 3 treatments with 10 replicates of 50 birds each. The experiment lasted for 6 wk, and dietary treatments included corn-soybean meal-based diet supplemented with 0, 0.01, and 0.02% GSE. The supplementation of GSE increased (P < 0.05) body weight gain (BWG) and final BW linearly but decreased (P < 0.05) feed-to-gain ratio (F/G) linearly during day (D) 22 to 42 and the entire experiment. The inclusion of GSE increased (P < 0.05) serum superoxide dismutase, glutathione peroxidase, total antioxidative capacity, catalase, complement4, immunoglobin G, interleukin-2, and interferon-γ linearly but decreased (P < 0.05) serum malondialdehyde linearly. The relative weight of carcass, breast meat, and spleen in GSE treatments was increased (P < 0.05) linearly, whereas the relative weight of abdominal fat was decreased linearly (P < 0.05). Birds fed GSE1 and GSE2 diets had lower (P < 0.05) cook loss, 2-thiobarbituric acid reactive substances, and drip loss on day 3 and 5 linearly but higher (P < 0.05) pH24h and water-holding capacity. The addition of GSE decreased (P < 0.05) jejunum crypt depth and ileal Escherichia coli counts linearly but increased (P < 0.05) jejunum villus height: crypt depth ratio and ileal Lactobacilli linearly. Taken together, the inclusion of GSE increased final BW and BWG, decreased F/G during day 22 to 42 and day 1 to 42, partially improved antioxidant activities, immunity, meat quality, and gut health in Pekin ducks.

Effects of dietary lipid sources on growth performance and carcass traits in Pekin ducks.

This study was conducted to determine the influence of dietary lipid sources on growth performance, carcass traits and taste scores in Pekin ducks. A total of 1,500 fifteen-day-old ducks (820 ± 22 g) were blocked based on body weight (BW), and randomly allotted to 3 treatments with 10 replicates of 50 birds each (25 males and 25 females). The experiment lasted for 4 wk, and dietary treatments included 3 different lipid sources (soybean oil, duck fat, and palm oil), which were evaluated in corn-soybean meal diets (3250 kcal/kg metabolizable energy and 16.5% crude protein for grower diet and 3350 kcal/kg metabolizable energy and 15.5% crude protein for finisher diet). During days 15 to 28, feeding soybean oil and palm oil diets increased (P < 0.05) body weight gain (BWG), but decreased (P < 0.05) feed intake, feed-to-gain ratio (F/G) and caloric conversion compared with duck fat. During days 29 to 42, birds fed duck fat diet had higher BWG, but lower (P < 0.05) F/G and caloric conversion than those fed soybean oil and palm oil diets. Overall, feeding soybean oil diet increased (P < 0.05) BWG and final BW, but decreased (P < 0.05) F/G compared with palm oil. Birds fed duck fat diet had higher (P < 0.05) skin, subcutaneous fat and abdominal fat yield compared with palm oil. Left breast meat yield in soybean oil group was higher (P < 0.05) than that in duck fat and palm oil groups. Birds in soybean oil group had lower (P < 0.05) roasting loss, but higher (P < 0.05) comprehensive score compared with duck fat and palm oil. In summary, birds fed soybean oil diet had the best growth performance and taste scores for roasting, whereas the duck fat was better in abdominal fat and subcutaneous fat yield than soybean oil and palm oil in Pekin ducks from 15 to 42 d of age under the same nutritional level.

Effects of astaxanthin produced by Phaffia rhodozyma on growth performance, antioxidant activities, and meat quality in Pekin ducks.

This study was conducted to determine the effect of astaxanthin (AX) produced by Phaffia rhodozyma (PR) on growth performance, antioxidant activities, relative organ weight, and meat quality in Pekin ducks. A total of 1,440 female one-day-old Pekin ducklings (52.3 ± 0.4 g) were blocked based on body weight (BW), and randomly allotted to 3 treatments with 8 replicates of 60 birds each. The experiment lasted for 6 wk, and dietary treatments included corn-soybean meal-based diet supplemented with 0, 0.15, and 0.3% PR. The supplementation of AX increased (P < 0.05) body weight gain (BWG) linearly and reduced (P < 0.05) feed-to-gain ratio (F/G) linearly during days 22 to 42. BWG and final BW was increased (P < 0.05) linearly by AX supplementation throughout the experiment. The inclusion of AX increased (P < 0.05) superoxide dismutase, glutathione peroxidase, total antioxidative capacity, and interleukin-6 in the serum linearly, as well as decreased (P < 0.05) serum malondialdehyde linearly. The relative weight of abdominal fat was increased (P < 0.05) linearly by AX supplementation. The inclusion of AX decreased (P < 0.05) cook loss linearly, but increased (P < 0.05) pH24h, water holding capacity and redness (a*) linearly. Taken together, the supplementation of AX (3.458 or 6.915 mg/kg diet) from PR improved final BW, BWG during days 22 to 42 and 1 to 42 and reduced F/G during days 22 to 42, as well as caused positive effects on antioxidant function and meat quality.

Effects of residual superdoses of phytase on growth performance, tibia mineralization, and relative organ weight in ducks fed phosphorus-deficient diets.

This study was conducted to determine the effects of residual superdoses of phytase on growth performance, tibia mineralization, and relative organ weight in ducks fed phosphorus-deficient diets. In Exp. 1, 4 kinds of commercial phytase were used to determine retention rate of phyatse with the phytase C being the highest via both high water-bath temperature (90%) and pelleting (50%), followed by phytase A, B, and D. In Exp. 2, a total of 560 male ducks were blocked based on body weight, and then allocated randomly to 7 treatments (5 replicates with 16 birds per replicate). Treatments included a maize-soybean meal-based diet with recommended calcium and 4.0 g non-phytate phosphorus (nPP)/kg starter diet or 3.8 g nPP/kg grower diet (positive control; PC), an nPP-deficient diet with 1.3 g nPP/kg starter diet or 1.1 g nPP/kg grower diet (negative control; NC), NC diets with increasing levels of residual phytase C (500, 1,000, 2,000, 3,000, and 4,000 units/kg feed) after pelleting. Birds fed NC diets had lower (P < 0.05) average daily gain (ADG) and average daily feed intake (ADFI) throughout the experiment compared with those fed PC diet. Supplementing NC diet with increasing residual superdoses of phytase improved (P < 0.05) ADG and ADFI quadratically in the entire experiment, while reduced feed-to-gain ratio (P < 0.05) quadratically during day 0 to 14. On day 14 and 35, birds fed NC diet had lower (P < 0.05) tibia length, weight, ash, calcium, phosphorus, and manganese contents than those fed PC diet. Increasing residual superdoses of phytase in NC diet increased (P < 0.05) tibia weight and ash, calcium, phosphorus contents quadratically on day 14 and 35. NC treatment increased (P < 0.05) the duodenum, jejunum, ileum, and cecum index compared with other treatments on day 14 and 35. Taken together, feeding increasing residual superdoses of phytase could counteract or exceed the negative effects of NC diet on growth performance, tibia mineralization, and relative organ weight in ducks.

Effects of maize naturally contaminated with aflatoxin B1 on growth performance, intestinal morphology, and digestive physiology in ducks.

This study was conducted to determine the effects of maize naturally contaminated with aflatoxin B1 (AFB1) on growth performance, intestinal morphology, relative digestive organs weight, digestive enzymes activities, and biochemical index of intestinal development in ducks. A total of 640 ducks was blocked on the basis of sex and body weight, and then allocated randomly to 2 treatments with 20 pens per treatment and 16 ducks per pen. The experiment lasted for 5 wk, and dietary treatments included basal diet (CON) and diets with 100% of normal maize replaced by AFB1 contaminated maize. Detectable levels of other toxins were present but only AFB1 exceeded limits and the level of AFB1 was 195.4 ug/kg in the contaminated maize, and ranged from 2.91 to 120.02 ug/kg in the starter diet and 2.03 to 153.12 ug/kg in the grower diet. Feeding AFB1 contaminated diets decreased (P < 0.05) ADG and ADFI during the whole experiment, whereas F/G during d 15 to 35 and d zero to 35 was reduced (P < 0.05). The mortality of ducks fed AFB1 contaminated diets increased (P < 0.05). Ducks fed AFB1 contaminated diets had greater (P < 0.05) relative weights of proventriculus and gizzard on d 14 as well as the duodenum, jejunum, and ileum on d 14 and 35. Feeding AFB1 contaminated diets increased (P < 0.05) crypt depth, villus width, and surface area in the duodenum on d 35 and villus height, villus width, and surface area in the jejunum on d 14. The activities of alkaline phosphatase and leucine aminopeptidase in the jejunum brush border together with chymotrypsin and trypsin in the pancreas increased (P < 0.05) on d 14 with the inclusion of AFB1 contaminated maize. The jejunum villus became long and wide in ducks fed AFB1 contaminated diets. Taken together, the feeding of maize naturally contaminated with AFB1 caused adverse effects on growth performance and intestinal morphology, and altered digestive physiology and development.

DEC1 regulates breast cancer cell proliferation by stabilizing cyclin E protein and delays the progression of cell cycle S phase.

Breast cancer that is accompanied by a high level of cyclin E expression usually exhibits poor prognosis and clinical outcome. Several factors are known to regulate the level of cyclin E during the cell cycle progression. The transcription factor DEC1 (also known as STRA13 and SHARP2) plays an important role in cell proliferation and apoptosis. Nevertheless, the mechanism of its role in cell proliferation is poorly understood. In this study, using the breast cancer cell lines MCF-7 and T47D, we showed that DEC1 could inhibit the cell cycle progression of breast cancer cells independently of its transcriptional activity. The cell cycle-dependent timing of DEC1 overexpression could affect the progression of the cell cycle through regulating the level of cyclin E protein. DEC1 stabilized cyclin E at the protein level by interacting with cyclin E. Overexpression of DEC1 repressed the interaction between cyclin E and its E3 ligase Fbw7α, consequently reducing the level of polyunbiquitinated cyclin E and increased the accumulation of non-ubiquitinated cyclin E. Furthermore, DEC1 also promoted the nuclear accumulation of Cdk2 and the formation of cyclin E/Cdk2 complex, as well as upregulating the activity of the cyclin E/Cdk2 complex, which inhibited the subsequent association of cyclin A with Cdk2. This had the effect of prolonging the S phase and suppressing the growth of breast cancers in a mouse xenograft model. These events probably constitute the essential steps in DEC1-regulated cell proliferation, thus opening up the possibility of a protein-based molecular strategy for eliminating cancer cells that manifest a high-level expression of cyclin E.

DACH1 inhibits SNAI1-mediated epithelial-mesenchymal transition and represses breast carcinoma metastasis.

Epithelial-mesenchymal transition (EMT) has a major role in cancer progression and metastasis. However, the specific mechanism of transcriptional repression involved in this process remains largely unknown. Dachshund homologue 1 (DACH1) expression is lost in invasive breast cancer with poor prognosis, and the role of DACH1 in regulating breast cancer metastasis is poorly understood. In this study, significant correlation between the expression of DACH1 and the morphology of breast cancer cells was observed. Subsequent investigation into the relationship between DACH1 and EMT showed that overexpression of DACH1 in ZR-75-30 cells induced a shift towards epithelial morphology and cell-cell adhesion, as well as increased the expression of the epithelial marker E-cadherin and suppressed cell migration and invasion. In contrast, silencing DACH1 in MCF-7 and T47D cells disrupted the epithelial morphology and cell-cell contact, reduced the expression of E-cadherin, and induced cell migration and invasion. DACH1 also specifically interacted with SNAI1, but not SNAI2, to form a complex, which could bind to the E-box on the E-cadherin promoter in an SNAI1-dependent manner. DACH1 inhibited the transcriptional activity of SNAI1, leading to the activation of E-cadherin in breast cancer cells. Furthermore, the level of DACH1 also correlated with the extent of metastasis in a mouse model. DACH1 overexpression significantly decreased the metastasis and growth of 4T1/Luc cells in BALB/c mice. Analysis of tissue samples taken from human breast cancers showed a significant correlation between the expression of DACH1 and E-cadherin in SNAI1-positive breast cancer. Collectively, our data identified a new mechanistic pathway for the regulation of EMT and metastasis of breast cancer cells, one that is based on the regulation of E-cadherin expression by direct DACH1-SNAI1 interaction.

Mammary-derived growth inhibitor (MDGI) cloned from human breast epithelial cells is expressed in fully differentiated lobular structures.

Mammary-derived growth inhibitor (MDGI), an inducer of rodent mammary differentiation and suppressor of human breast cancer cell growth, has been cloned from bovine and rodent mammary glands. The present study reports the cloning of MDGI from cultured human breast epithelial cells (HBEC-MDGI) as a cDNA fragment encoding a protein of 133 amino acids identical to heart fatty acid binding protein. Expression of HBEC-MDGI, as detected by in situ hybridization in paraffin-embedded normal breast tissues, was maximal in the most differentiated lobules type 4, low in the moderately differentiated lobules type 3, and absent in the least differentiated lobules types 1 and 2. HBEC-MDGI was not expressed in breast tissues that contained ductal hyperplasia, carcinoma in situ or invasive carcinomas. Our results indicate that HBEC-MDGI is a biomarker of lobular differentiation in the human breast, and its expression is silenced in poorly differentiated lobules as well as in the early and late stages of breast cancer progression.

S100P calcium-binding protein overexpression is associated with immortalization of human breast epithelial cells in vitro and early stages of breast cancer development in vivo.

The mechanism of cell immortalization of human breast epithelial cells leading to neoplastic transformation is not clear. The isolation and characterization of a spontaneously immortalized human breast epithelial cell line, MCF-10F, have provided a valuable tool to identify genes involved in this process. Using the technique of differential display, we have identified seven cDNA bands differentially displayed in the MCF-10F cells when compared with the mortal S130 cells from which MCF-10F was originated. One of these bands was isolated and cloned. Sequence analysis revealed 99% homology to the EF-hand calcium-binding protein S100P (Placental). The clone was overexpressed in the immortal cell line MCF-10F when compared to the mortal counterpart S130 or other primary cultures of human breast epithelial cells. In addition, it was highly expressed in chemically transformed breast epithelial cell lines (BP1E and D3. 1), breast cancer cell line T47D, as well as in three invasive ductal carcinomas when compared to their normal adjacent tissue. The S100P protein was localized by immunohistochemistry, using a monoclonal antibody against the same amino acid sequence of the gene cloned, in ductal hyperplasias, in situ and invasive ductal carcinoma, but not in the normal tissues. We concluded that S100P overexpression is an early event that might play an important role in the immortalization of human breast epithelial cells in vitro and tumor progression in vivo.

Frequent estrogen and progesterone receptor immunoreactivity in renal angiomyolipomas from women with pulmonary lymphangioleiomyomatosis.

OBJECTIVE: To determine whether renal angiomyolipomas from women with pulmonary lymphangioleiomyomatosis (LAM) express estrogen receptor (ER) and progesterone receptor (PR). DESIGN: Retrospective study of archival tissue. PATIENTS: Twelve women with LAM and angiomyolipomas. SETTING: Fox Chase Cancer Center. INTERVENTIONS: ER and PR expression was studied using immunohistochemistry. The hormonal status of the patients at the time of resection of the angiomyolipoma was determined. RESULTS: Ten of the angiomyolipomas had ER immunoreactivity (83%), and all 12 had PR immunoreactivity (100%). The ER and PR positivity was in the smooth muscle component of the angiomyolipomas only. For five women, pulmonary LAM specimens were also available; two were ER positive (40%), and all five were PR positive (100%). All four angiomyolipomas from women receiving progesterone therapy were ER and PR positive. One tumor from a woman receiving tamoxifen was ER negative and strongly PR positive. One woman was pregnant; her tumor was ER and PR positive. CONCLUSIONS: ER and PR expression is frequent in renal angiomyolipoma cells from women with LAM. PR was more consistently present than ER in angiomyolipomas and in LAM. Our data suggest that angiomyolipoma growth could be affected by hormonal factors. If the growth of LAM-associated angiomyolipomas slows during hormonal therapy, there are two potential implications for LAM patients: first, angiomyolipoma size could serve as a measurable indication of response to hormonal therapy; and second, surgical removal of angiomyolipomas might be avoided in some cases.

A study of equivalent source techniques for high-resolution EEG imaging.

High-resolution EEG imaging has been an important topic in recent EEG research, and much work has been done on the two equivalent source imaging techniques: the equivalent distributed dipole-layer source imaging technique (EST) and the equivalent multipole source imaging technique (SAT). In this paper we first develop a forward density formula for a spherical equivalent distributed dipole layer of an arbitrary dipole in a three-concentric-sphere head model. It is clarified using the derived forward formula that the equivalent dipole-layer source and equivalent multipole source are interrelated in theory. Finally, simulation comparisons are conducted, the results of which suggest that EST has a higher spatial resolution than SAT when both of them are implemented by a truncated singular value decomposition algorithm. This is due to the different singularities of the inversion equations involved in the two techniques. An empirical VEP data study also shows that EST is better than SAT in providing higher spatial resolution EEG imaging.

Spectrophotometric study on the supramolecular interactions of nile blue sulphate with nucleic acids.

The supramolecular interaction of nile blue sulphate (NBS) with nucleic acids was studied by investigating the characteristics of the interaction absorption spectra on the basis of the drug binding process in organic system in which small amount of drug interacting with large amount of biological macromolecules involves, and an accordingly binding model for organic dyes with large amount of macromolecules was established. At pH 7.40 and ionic strength 0.004, the H-aggregation of NBS occurs with increasing NBS concentration. The NBS aggregates can be bound to both calf thymus DNA and fish sperm DNA by the ratio of each nucleotide residue with a molecule of NBS if the concentration of DNAs is more than 15-fold excessive. The corresponding binding constant for the interaction of NBS with DNAs is about 10(3) order, with which thermodynamic parameters for the interactions, such as the change of free energy, enthalpy and entropy at 25 degrees C, were calculated. It was found that the binding of NBS with thermally denatured DNA is similar to that with native yeast RNA, which indicates H-aggregation of NBS can be encouraged by single stranded nucleic acids.

Regulation of autophagy by the Rab GTPase network.

Autophagy (macroautophagy) is a highly conserved intracellular and lysosome-dependent degradation process in which autophagic substrates are enclosed and degraded by a double-membrane vesicular structure in a continuous and dynamic vesicle transport process. The Rab protein is a small GTPase that belongs to the Ras-like GTPase superfamily and regulates the vesicle traffic process. Numerous Rab proteins have been shown to be involved in various stages of autophagy. Rab1, Rab5, Rab7, Rab9A, Rab11, Rab23, Rab32, and Rab33B participate in autophagosome formation, whereas Rab9 is required in non-canonical autophagy. Rab7, Rab8B, and Rab24 have a key role in autophagosome maturation. Rab8A and Rab25 are also involved in autophagy, but their role is unknown. Here, we summarize new findings regarding the involvement of Rabs in autophagy and provide insights regarding future research on the mechanisms of autophagy regulation.

CLOCK is a substrate of SUMO and sumoylation of CLOCK upregulates the transcriptional activity of estrogen receptor-α.

Disruption of the circadian rhythm is now believed to associate with a number of hormone-related cancers, such as breast cancer, in which aberrant estrogen receptor-α (ERα) signaling is a major contributor. However, the molecular mechanisms underlying the function of core clock proteins in cancer are still largely undefined. In this study, we showed that circadian locomotor output cycles kaput (CLOCK), a key circadian protein, can interact with ERα. Furthermore, this interaction was enhanced by estrogen. We also showed that CLOCK can be sumoylated and sumoylation of CLOCK, which is also stimulated by estrogen, had two consequences: (1) it increased the transcriptional activity of CLOCK; and (2) it increased the CLOCK-modulated transcriptional activity of ERα, as shown by increased transcription of cyclin D1. Sumoylation of CLOCK occurred at two lysine residues, K67 and K851. The enhancement of ERα transcriptional activity exerted by wild-type but not mutant (2K/2R) CLOCK in response to estrogen indicated that sumoylation of CLOCK may have an important role in estrogen-dependent signaling. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay conducted with breast cancer cell lines (MCF-7 and T47D) demonstrated that sumoylation of CLOCK stimulated cell growth and increased the proportion of S phase cells in the cell cycle. The results of this study uncovered new insight into the connection between a major circadian protein and a major estrogen-dependent transcription factor, providing the basis for further research into the involvement of circadian proteins in breast cancer.