A Novel sLRP6E1E2 Inhibits Canonical Wnt Signaling, Epithelial-to-Mesenchymal Transition, and Induces Mitochondria-Dependent Apoptosis in Lung Cancer

Aberrant activation of the Wnt pathway contributes to human cancer progression. Antagonists that interfere with Wnt ligand/receptor interactions can be useful in cancer treatments. In this study, we evaluated the therapeutic potential of a soluble Wnt receptor decoy in cancer gene therapy. We designed a Wnt antagonist sLRP6E1E2, and generated a replication-incompetent adenovirus (Ad), dE1-k35/sLRP6E1E2, and a replication-competent oncolytic Ad, RdB-k35/sLRP6E1E2, both expressing sLRP6E1E2. sLRP6E1E2 prevented Wnt-mediated stabilization of cytoplasmic β-catenin, decreased Wnt/β-catenin signaling and cell proliferation via the mitogen-activated protein kinase, and phosphatidylinositol 3-kinase pathways. sLRP6E1E2 induced apoptosis, cytochrome c release, and increased cleavage of PARP and caspase-3. sLRP6E1E2 suppressed growth of the human lung tumor xenograft, and reduced motility and invasion of cancer cells. In addition, sLRP6E1E2 upregulated expression of epithelial marker genes, while sLRP6E1E2 downregulated mesenchymal marker genes. Taken together, sLRP6E1E2, by inhibiting interaction between Wnt and its receptor, suppressed Wnt-induced cell proliferation and epithelial-to-mesenchymal transition

Cold shock domain proteins and glycine-rich RNA-binding proteins from Arabidopsis thaliana can promote the cold adaptation process in Escherichia coli

Despite the fact that cold shock domain proteins (CSDPs) and glycine-rich RNA-binding proteins (GRPs) have been implicated to play a role during the cold adaptation process, their importance and function in eukaryotes, including plants, are largely unknown. To understand the functional role of plant CSDPs and GRPs in the cold response, two CSDPs (CSDP1 and CSDP2) and three GRPs (GRP2, GRP4 and GRP7) from Arabidopsis thaliana were investigated. Heterologous expression of CSDP1 or GRP7 complemented the cold sensitivity of BX04 mutant Escherichia coli that lack four cold shock proteins (CSPs) and is highly sensitive to cold stress, and resulted in better survival rate than control cells during incubation at low temperature. In contrast, CSDP2 and GRP4 had very little ability. Selective evolution of ligand by exponential enrichment (SELEX) revealed that GRP7 does not recognize specific RNAs but binds preferentially to G-rich RNA sequences. CSDP1 and GRP7 had DNA melting activity, and enhanced RNase activity. In contrast, CSDP2 and GRP4 had no DNA melting activity and did not enhance RNAase activity. Together, these results indicate that CSDPs and GRPs help E.coli grow and survive better during cold shock, and strongly imply that CSDP1 and GRP7 exhibit RNA chaperone activity during the cold adaptation process

Pilot-Scale Lactic Acid Production via Batch Culturing of Lactobacillus sp. RKY2 Using Corn Steep Liquor As a Nitrogen Source

U ovom se istraživanju pokušala odrediti učinkovitost proizvodnje mliječne kiseline uzgojem soja Lactobacillus sp. RKY2 u pilot-postrojenju uporabom kukuruznog ekstrakta kao izvora dušika. Količina mliječne kiseline nakon fermentacije, rast stanica, prinos i produktivnost nisu se bitno promijenili dodatkom čiste glukoze pri povećanju obujma procesa s 2,5 na 30 i 300 L. U svim pokusima udio mliječne kiseline linearno se povećavao s porastom početne koncentracije glukoze. U pokusu s hidrolizatom drva smanjili su se produktivnost mliječne kiseline i rast stanica s povećanjem obujma procesne opreme, zbog toksičnih kemikalija iz hidrolizata. Međutim, u svim je pokusima prinos mliječne kiseline bio veci od 90 % bez obzira na utrošak glukoze. Zaključeno je da je mliječna kiselina uspješno proizvedena u pilot-postrojenju upotrijebljenom u ovom istraživanju.In this study, the determination of the efficiency of a pilot-scale fermentation process using corn steep liquor as a nitrogen source was attempted in order to produce lactic acid via batch culturing of Lactobacillus sp. RKY2. Using pure glucose, fermentation efficiency characteristics, such as final lactic acid, cell growth, yield, and productivity were not substantially influenced by the scale-up of the laboratory-scale fermentation from 2.5- to 30- and 300-litre scale fermentations. In all experiments, the content of lactic acid produced increased in a linear fashion with increases in the initial glucose concentration. In the experiments using wood hydrolyzate, both lactic acid productivity and cell growth were decreased as a result of the scaling-up of the fermentation. This might be attributed to the toxic chemicals contained in the wood hydrolyzates. However, in all experiments, lactic acid yields remained higher than 90 % with regard to the amount of glucose consumed. Therefore, lactic acid was successfully produced by the pilot-scale bioreactor scheme adopted in this study

Pilot-Scale Lactic Acid Production via Batch Culturing of Lactobacillus sp. RKY2 Using Corn Steep Liquor As a Nitrogen Source

U ovom se istraživanju pokušala odrediti učinkovitost proizvodnje mliječne kiseline uzgojem soja Lactobacillus sp. RKY2 u pilot-postrojenju uporabom kukuruznog ekstrakta kao izvora dušika. Količina mliječne kiseline nakon fermentacije, rast stanica, prinos i produktivnost nisu se bitno promijenili dodatkom čiste glukoze pri povećanju obujma procesa s 2,5 na 30 i 300 L. U svim pokusima udio mliječne kiseline linearno se povećavao s porastom početne koncentracije glukoze. U pokusu s hidrolizatom drva smanjili su se produktivnost mliječne kiseline i rast stanica s povećanjem obujma procesne opreme, zbog toksičnih kemikalija iz hidrolizata. Međutim, u svim je pokusima prinos mliječne kiseline bio veci od 90 % bez obzira na utrošak glukoze. Zaključeno je da je mliječna kiselina uspješno proizvedena u pilot-postrojenju upotrijebljenom u ovom istraživanju.In this study, the determination of the efficiency of a pilot-scale fermentation process using corn steep liquor as a nitrogen source was attempted in order to produce lactic acid via batch culturing of Lactobacillus sp. RKY2. Using pure glucose, fermentation efficiency characteristics, such as final lactic acid, cell growth, yield, and productivity were not substantially influenced by the scale-up of the laboratory-scale fermentation from 2.5- to 30- and 300-litre scale fermentations. In all experiments, the content of lactic acid produced increased in a linear fashion with increases in the initial glucose concentration. In the experiments using wood hydrolyzate, both lactic acid productivity and cell growth were decreased as a result of the scaling-up of the fermentation. This might be attributed to the toxic chemicals contained in the wood hydrolyzates. However, in all experiments, lactic acid yields remained higher than 90 % with regard to the amount of glucose consumed. Therefore, lactic acid was successfully produced by the pilot-scale bioreactor scheme adopted in this study

Substance P and beta-endorphin mediate electro-acupuncture induced analgesia in mouse cancer pain model

<p>Abstract</p> <p>Background</p> <p>Opioid analgesics are generally used to combat the pain associated with cancerous conditions. These agents not only inhibit respiratory function and cause constipation, but also induce other significant side effects such as addiction and tolerance, all of which further contribute to a reduced quality of life for cancer patients. Thus, in the present study, the effects of electro-acupuncture treatment (EA) on mechanical allodynia were examined in a cancer pain mouse model.</p> <p>Methods</p> <p>In order to produce a neuropathic cancer pain model, S-180 sarcoma cells were inoculated around the sciatic nerve of left legs of Balb/c mice. Magnetic Resonance Imaging (MRI) scanning confirmed the mass of S-180 cancer cells embedded around the sciatic nerve. Mechanical allodynia was most consistently induced in the mouse sarcoma cell line S-180 (2 × 10⁶sarcoma cells)-treated group compared to all the other groups studied. EA stimulation (2 Hz) was administered daily to ST36 (Zusanli) of S-180 bearing mice for 30 min for 9 days after S-180 inoculation.</p> <p>Results</p> <p>EA treatment significantly prolonged paw withdrawal latency from 5 days after inoculation. It also shortened the cumulative lifting duration from 7 days after inoculation, compared to the tumor control. Also, the overexpression of pain peptide substance P in the dorsal horn of the spinal cord was significantly decreased in the EA-treated group compared to the tumor control on Day 9 post inoculation. Furthermore, EA treatment effectively increased the concentration of β-endorphin in blood and brain samples of the mice to a greater extent than that of the tumor control as well as the normal group. The concentration of β-endorphin for EA treatment group increased by 51.457% in the blood and 12.6% in the brain respectively, compared to the tumor control group.</p> <p>Conclusion</p> <p>The findings of this study suggest that a S-180 cancer pain model is useful as a consistent and short time animal model. It also indicated that EA treatment could be used as an alternative therapeutic method for cancer pain due to a consequent decrease in substance P and increase in β-endorphin levels.</p

Alcohol induces cell proliferation via hypermethylation of ADHFE1 in colorectal cancer cells

BACKGROUND: The hypermethylation of Alcohol dehydrogenase iron containing 1 (ADHFE1) was recently reported to be associated with colorectal cancer (CRC) differentiation. However, the effect of alcohol on ADHFE1 hypermethylation in CRC is still unclear. METHODS: The methylation status and expression levels of ADHFE1 were investigated in primary tumor tissues and adjacent normal tissues of 73 patients with CRC, one normal colon cell line, and 4 CRC cell lines (HT-29, SW480, DLD-1, and LoVo) by quantitative methylation-specific polymerase chain reaction (QMSP) and real-time reverse transcription polymerase chain reaction (real time PCR), respectively. The effect of alcohol on the methylation status of ADHFE1 was analyzed in HT-29, SW480, DLD-1, and CCD18Co cells using QMSP, real-time PCR, immunoblot, and cell proliferation assay. RESULTS: ADHFE1 was hypermethylated in 69 of 73 CRC tissues (95%) compared to adjacent normal tissues (p < 0.05). The mRNA expression of ADHFE1 was significantly reduced in CRC compared to adjacent normal tissues (p < 0.05) and its expression was decreased in the alcohol consumption group (p < 0.05). ADHFE1 was hypermethylated and its expression was decreased in 4 CRC cell lines compared with normal colon cell line. Alcohol induced hypermethylation of ADHFE1, decreased its expression, and stimulated cell proliferation of HT-29, SW480, and DLD-1cells. CONCLUSION: These results demonstrate that the promoter hypermethylation of ADHFE1 is frequently present in CRC and alcohol induces methylation-mediated down expression of ADHFE1 and proliferation of CRC cells

Left ventricular dysfunction measured by tissue Doppler imaging and strain rate imaging in hypertensive adolescents

Purpose : Left ventricular (LV) hypertrophy and impaired diastolic function may occur early in systemic hypertension. Diastolic dysfunction is associated with increased cardiovascular risk. Tissue Doppler imaging (TDI)-derived tissue velocity and strain rate are new parameters for assessing diastolic dysfunction. The aim of this study is to determine whether TDI and strain rate imaging (SRI) would improve the ability to recognize early impaired diastolic and systolic functions compared with conventional echocardiography in hypertensive adolescents. Methods : We included 38 hypertensive patients with systolic blood pressure above 140 mmHg or diastolic blood pressure above 90 mmHg. Ejection fraction and myocardial performance index (MPI) were estimated by conventional echocardiography. Peak systolic myocardial velocity, early diastolic myocardial velocity (Em), and peak late diastolic myocardial velocity (Am) were obtained by using TDI and SRI. Results : In the hypertensive group, interventricular septal thickness was significantly increased on M-mode echocardiography. Em&#47;Am was significantly decreased at the mitral valve annulus. Among hypertensive subjects, the E strain rate at basal, mid, and apex was significantly decreased. Systolic strain was significantly decreased at the septum in the hypertensive group. Conclusion : Strain rate might be a useful new parameter for the quantification of both regional and global LV functions and could be used in long-term follow up in hypertensive patients. Early identification by SRI of subjects at risk for hypertensive and ventricular dysfunction may help to stratify risk and guide therapy. Further studies, including serial assessment of LV structure and function in a larger number of adolescents with hypertension, is necessary