OBJECTIVE: Retinal dehydrogenase 1 (RALDH1) is a cytosolic enzyme which acts both as a source of retinoic acid (RA) and as a detoxification enzyme. RALDH1 has key functions in the midbrain dopaminergic system, which influences motivation, cognition, and social behavior. Since dopamine has been increasingly linked to autism spectrum disorder (ASD), we asked whether RALDH1 could contribute to the autistic phenotype. Therefore, we investigated for the first time the levels of RALDH1 in autistic patients. To further assess the detoxification function of RALDH1, we also explored 4-hydroxynonenal protein adducts (4-HNE PAs) and reduced glutathione (GSH) levels. Moreover, considering the effect of testosterone on RALDH1 expression, we measured the second to fourth digit ratio (2D:4D ratio) for both hands, which reflects exposure to prenatal testosterone. METHODS: Male patients with ASD (n=18; age, 62.9±4.3 months) and healthy controls (n=13; age, 78.1±4.9 months) were examined. Erythrocyte RALDH1, serum 4-HNE PAs and erythrocyte GSH levels were measured using colorimetric assays, and digit lengths were measured using digital calipers. RESULTS: We found significantly lower (−42.9%) RALDH1 levels in autistic patients as compared to controls (p=0.032). However, there was no difference in 4-HNE PAs levels (p=0.368), GSH levels (p=0.586), or 2D:4D ratios (p=0.246 in the left hand, p=0.584 in the right hand) between healthy controls and autistic subjects. CONCLUSION: We concluded that a subset of autistic patients had a low RALDH1 level. These results suggest that low RALDH1 levels could contribute to the autistic phenotype by reflecting a dopaminergic dysfunction.
Autism Spectrum Disorder* ; Autistic Disorder* ; Cognition ; Cytosol ; Dopamine* ; Erythrocytes ; Glutathione ; Hand ; Humans ; Male ; Mesencephalon ; Motivation ; Phenotype ; Retinal Dehydrogenase* ; Retinaldehyde* ; Social Behavior ; Testosterone ; Tretinoin

PURPOSE: To investigate the effects of anti-vascular endothelial growth factor (VEGF) antibody on the survival of retinal ganglion cell (RGC)-5 cells differentiated with staurosporine under oxidative stress. METHODS: We used real-time polymerase chain reaction and Western blot to confirm the expression of VEGF, VEGF receptor (VEGFR)-1 and VEGFR-2 in RGC-5 cells differentiated with staurosporine for 6 hours. The differentiated RGC-5 cells were treated with 800 µM hydrogen peroxide (H₂O₂) for 24 hours to induce oxidative stress. Then, the survival rate of RGC-5 was confirmed by lactate dehydrogenase assay at each concentration (0, 0.01, 0.1, and 1 mg) using bevacizumab as the anti-VEGF antibody. The expression of VEGF, VEGFR-1, and VEGFR-2 was confirmed using real-time polymerase chain reaction. RESULTS: VEGF, VEGFR-1, and VEGFR-2 were all expressed in differentiated RGC-5 cells. When RGC-5 cells were simultaneously treated with bevacizumab and 800 µM H₂O₂, survival of RGC-5 decreased with bevacizumab concentration. VEGF expression in RGC-5 cells increased with increasing concentration of bevacizumab. Similar patterns were observed for VEGFR-1 and VEGFR-2, but the degree of increase was smaller than that for VEGF. CONCLUSIONS: When bevacizumab was administered to differentiated RGC-5 cells, the cell damage caused by oxidative stress increased. Therefore, given these in vitro study results, caution should be exercised with bevacizumab treatment.
Bevacizumab ; Blotting, Western ; Endothelial Growth Factors* ; Hydrogen Peroxide ; In Vitro Techniques ; L-Lactate Dehydrogenase ; Oxidative Stress ; Real-Time Polymerase Chain Reaction ; Receptors, Vascular Endothelial Growth Factor ; Retinal Ganglion Cells* ; Retinaldehyde* ; Staurosporine ; Survival Rate ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor Receptor-1 ; Vascular Endothelial Growth Factor Receptor-2

OBJECTIVETo investigate the expression of ALDH1, CXCR4 and E-cadherin in gastric carcinoma and their roles in lymphatic metastasis.

METHODSSurgical specimens from 127 cases of gastric carcinoma were examined for expressions of ALDH1, CXCR4 and E-cadherin immuohistochemistry with 60 adjacent tissues as control. The associations of ALDH1, CXCR4 and E-cadherin with the clinicopathological pfeatures, 5-year survival rate and lymph node metastasis of the patients were analyzed.

RESULTSALDH1, CXCR4 and E-cadherin were positive in 57.5% (73/127), 63.8% (81/127), and 36.2% (46/127) of the gastric carcinoma tissues, respectively, showing significant differences from the rates in the adjacent tissues (P<0.05). The expression of ALDH1 was significantly correlated with TNM stage and lymph node metastasis (P<0.05), CXCR4 was significantly correlated with the invasion depth, differentiation, TNM stage and lymph node metastasis of the tumor (P<0.05), and E-cadherin was significantly correlated with the invasion depth, differentiation and lymph node metastasis (P<0.05). The positivity rates of ALDH1, CXCR4 and E-cadherin were higher in cases with lymph node metastasis than in those without metastasis. E-cadherin expression was inversely correlated with ALDH1 and CXCR4 expression, and the latter two were positively correlated (P<0.001). Overexpressions of ALDH1 and CXCR4 and a decreased expression of E-cadherin were all related to a poor prognosis of the patients (P<0.05). The expressions ofALDH1, CXCR4 and E-cadherin were all independent prognostic factors of gastric carcinoma.

CONCLUSIONThe expressions of ALDH1, CXCR4 and E-cadherin are associated with the invasion, metastasis and prognosis of gastric carcinoma, and their combined detection provides important evidence for predicting the progression and prognosis of gastric carcinoma.

Cadherins ; genetics ; metabolism ; Carcinoma ; genetics ; metabolism ; Disease Progression ; Humans ; Isoenzymes ; genetics ; metabolism ; Lymphatic Metastasis ; Prognosis ; Receptors, CXCR4 ; genetics ; metabolism ; Retinal Dehydrogenase ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism ; Survival Rate

Previous studies have demonstrated that spheroid type cells grown under suspension culture conditions have cancer stem cell (CSC) traits in a number of cancers, but this phenomenon has not yet been reported in the VX2 rabbit oral cancer model. Hence, this study aimed to study the spheroid cells from VX2 rabbit buccal squamous cell carcinomas (SCCs) and assess their CSC characteristics. Five adult male New Zealand white outbred rabbits were used to generate VX2 rabbit buccal SCC. Sphere-forming cell culture was performed for the VX2 rabbit buccal SCC specimens. The self-renewal capability; cluster of designation (CD) 44, CD133, acetaldehyde dehydrogenase 1 (ALDH1), B cell-specific Moloney murine leukemia virus integration site 1 (Bmi-1), Nestin, octamer-binding transcription factor 4 (Oct4) and reduced expression protein-1 (Rex-1) expression with reverse transcription-polymerase chain reaction (RT-PCR); chemoresistance to cisplatin and 5-fluorouracil; and in vivo tumorigenicity of spheroid cell transplantation in nude mice were evaluated to determine the CSC characteristics of the resulting spheroid cells. We successfully obtained spheroid cells from the VX2 rabbit OSCC tissues. The spheroid cells exhibited CSC traits, including the expression of CSC and stem cell markers (CD44, Bmi-1, Nestin, Oct4 and Rex-1), capacity to generate new spheroid colonies within 1 week of reseeding from single-dissociated spheroid cells, chemoresistance capacity and generation of tumour xenografts (with histological features resembling those of the original VX2 rabbit buccal SCC) from the transplantation of 10(3) undifferentiated spheroid cells into nude mice. In summary, we demonstrated that spheroid cells with CSC cell traits can be derived from VX2 rabbit buccal SCCs, indicating that this animal cancer model is applicable for studying CSCs in human oral cancers.
AC133 Antigen ; Animals ; Antigens, CD ; analysis ; Antineoplastic Agents ; pharmacology ; Carcinoma, Squamous Cell ; pathology ; Cell Culture Techniques ; Cisplatin ; pharmacology ; DNA-Binding Proteins ; analysis ; Disease Models, Animal ; Drug Resistance, Neoplasm ; Fluorouracil ; pharmacology ; Glycoproteins ; analysis ; Heterografts ; transplantation ; Hyaluronan Receptors ; analysis ; Isoenzymes ; analysis ; Male ; Mice ; Mice, Nude ; Mouth Neoplasms ; pathology ; Neoplasm Transplantation ; Neoplastic Stem Cells ; classification ; Nestin ; analysis ; Octamer Transcription Factor-3 ; analysis ; Peptides ; analysis ; Polycomb Repressive Complex 1 ; analysis ; Proto-Oncogene Proteins ; analysis ; Rabbits ; Retinal Dehydrogenase ; analysis ; Spheroids, Cellular ; classification

AC133 Antigen ; Animals ; Antigens, CD ; metabolism ; Biomarkers, Tumor ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; CD24 Antigen ; metabolism ; CD55 Antigens ; metabolism ; Female ; Gangliosides ; metabolism ; Glycoproteins ; metabolism ; Hedgehog Proteins ; metabolism ; Humans ; Hyaluronan Receptors ; metabolism ; Isoenzymes ; metabolism ; Neoplastic Stem Cells ; metabolism ; Octamer Transcription Factor-3 ; metabolism ; Peptides ; metabolism ; Receptors, Notch ; metabolism ; Retinal Dehydrogenase ; metabolism ; Signal Transduction ; Wnt Signaling Pathway

Abstract: The first-line drug artemisinin is widely used against malaria. Commercially available artemisinin is extracted from plants. However, the lack of sufficient raw material, artemisinin and the cost associated with the drug's manufacture have limited the supply of ACT to most malaria sufferers in the Developing World. As such, it is important to develop a low cost, fine to environment and high-quality method to supply sufficient and reliable quantities of artemisinin in the future. The field of synthetic biology, which utilizes cell factories to manipulate microbial metabolism to enhance the production of artemisinin and its intermediates, has a particularly strong impact by providing new platforms for chemical production. After a brief introduction of the artemisinin biosynthetic pathway, the present review focuses on the introduction of artemisinin biosynthetic genes, such as the genes encoding amorpha-4, 11-diene monooxygenase, NADPH: cytochrome P450 oxidoreductase, artemisinic aldehyde delta 11(13) reductase and aldehyde dehydrogenase. The review also addresses general considerations for potential contributions of synthetic biology to artemisinin production, with an emphasis on factors influencing interest compounds production in chassis cells.
Antimalarials ; metabolism ; supply & distribution ; Artemisinins ; metabolism ; supply & distribution ; Biosynthetic Pathways ; Cytochrome P-450 Enzyme System ; genetics ; Escherichia coli ; metabolism ; Gene Dosage ; Genetic Engineering ; Isoenzymes ; genetics ; RNA Nucleotidyltransferases ; genetics ; Retinal Dehydrogenase ; genetics ; Saccharomyces cerevisiae ; metabolism ; Synthetic Biology

OBJECTIVETo investigate the expression pattern of CD133 and ALDH1 in colorectal cancer cells line Colo205 cultured in serum-free medium (SFM) containing recombinant human epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF).

METHODSColo205 cells were cultured in serum-free medium (SFM) containing human recombinant EGF and bFGF or in serum-supplemented medium (SSM). The expression of CD133 was analyzed in both groups, and CD133(+) and CD133(-) cells sorted from the SFM group using flow cytometry and observed microscopically for their growth status. The expression of CD133 and ALDH1 in CD133(+) cells and CD133(-) cells was detected by immunofluorescence assay. CD133(+) cells and CD133(-) cells were then injected subcutaneously into NOD/SCID mice and the expression of ALDH1 in the tumor tissues was detected by immunohistochemistry.

RESULTSThe cells in SFM group showed a significantly higher percentage of CD133(+) cells than those in SSM group (P<0.05). In SFM, CD133(+) cells were capable of forming tumor spheres while CD133(-) cells could not; CD133(+)cells strongly expressed CD133 and ALDH1 and CD133(-) cells did not. In mice, tumors generated by CD133(+) cells, but not by CD133(-) cells, positively expressed ALDH1.

CONCLUSIONSCD133(+) Colo205 colorectal cancer cells in SFM containing human recombinant EGF and bFGF can form tumor spheres and strongly express ALDH1. ALDH1 may be one of the candidate markers of colorectal cancer stem cells.

AC133 Antigen ; Animals ; Antigens, CD ; metabolism ; Cell Culture Techniques ; Cell Line, Tumor ; Colorectal Neoplasms ; metabolism ; Culture Media, Serum-Free ; Glycoproteins ; metabolism ; Humans ; Isoenzymes ; metabolism ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Peptides ; metabolism ; Retinal Dehydrogenase ; metabolism

OBJECTIVE: To determine the apoptosis-inducing effect of ultraviolet light (UV) on human lens epithelial cell (HLEC) and to explore the involvement of changes in ALDH1 folowing UV radiation. METHODS: HLEC was exposed to the same UV light source and was subsequently divided into 6 groups according to UV radiation time of 0 (control group), 5, 10, 15, and 30 min. Apoptosis was detected by AO/EB staining. Changes of ALDH1 in HLEC were detected by immunohistochemical staining and Western blot. RESULTS: The intensity of immunohistochemical staining and the rate of positive cells decreased with increase of UV time (P<0.05). The rate of positive ALDH1 cells was negatively correlated with the rate of apoptosis (r= -0.92, P<0.05). Western blot showed the integrated absorbance values significantly decreased with the increase of UV time (P<0.05). CONCLUSION ALDH1 in HLEC decreases with an increase of UV exposure, which may be related to UV induced apoptosis of HLEC.
Aldehyde Dehydrogenase 1 ; Apoptosis ; radiation effects ; Cells, Cultured ; Epithelial Cells ; cytology ; metabolism ; radiation effects ; Humans ; Isoenzymes ; genetics ; metabolism ; Lens, Crystalline ; cytology ; Retinal Dehydrogenase ; genetics ; metabolism ; Ultraviolet Rays ; adverse effects

The cancer stem cell (CSC) hypothesis proposes that CSCs are the root of cancer. CSC-targeted therapies may prevent cancer relapse and provide more effective treatment. The expression of aldehyde dehydrogenase 1, as assessed by the Aldefluor assay, has been recognized as a marker of CSCs in breast cancer. Inhibitors of DNA-binding proteins (IDs) have an important role in stem cell differentiation. In this study, we examined IDs necessary for the regulation of stem properties in Aldefluorpos 4T1 cells. When the expression profile of IDs in Aldefluorneg and Aldefluorpos 4T1 cells was compared, qRT-PCR analysis showed that ID4 expression was highly upregulated in Aldefluorpos 4T1 cells. In addition, knockdown of ID4 expression suppressed the properties of CSCs, including their sphere-forming ability and side population phenotype. The findings suggest that ID4 may be a therapeutic target for the treatment of advanced breast cancer.
Aldehyde Dehydrogenase ; Animals ; Breast Neoplasms ; Cell Line ; DNA-Binding Proteins ; Isoenzymes ; Mice ; Neoplastic Stem Cells ; Phenotype ; Recurrence ; Retinal Dehydrogenase ; Stem Cells

OBJECTIVETo isolate breast cancer stem cells from breast cancer patients and identify their biological characteristics.

METHODSMammospheric cells were purified and enriched from the tumor tissues of breast cancer patients using mammosphere culture. Their expressions of CD44 and CD24 were analyzed by flow cytometry, and ALDH1, ESA and Oct4 expressions were determined by Western Blotting. The primary mammospheric and adherent cells, at the density of 2×10(4), 2×10(5) or 2×10(6), were inoculated into NOD/SCID mice to observe their tumorigenic and metastatic activities.

RESULTSWith mammosphere culture method, 62.36% of the mammospheric cells showed CD44(+)/CD24(-/low) phenotype. The expressions of ALDH1, ESA and Oct4 in the mammospheric cells were significantly higher than those in the adherent culture-derived breast cancer cells (P<0.05). Primary mammospheric cells were at least 100-fold more tumorigenic than the adherent cells; the mammospheric cells were associated with liver or lung metastases, but the adherent cells were not.

CONCLUSIONMammosphere culture can be employed to obtain breast cancer stem cells from the tumor tissues of breast cancer patients.

Adult ; Animals ; Aryldialkylphosphatase ; metabolism ; Breast Neoplasms ; pathology ; CD24 Antigen ; metabolism ; Cell Culture Techniques ; methods ; Female ; Humans ; Hyaluronan Receptors ; metabolism ; Isoenzymes ; metabolism ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Middle Aged ; Neoplastic Stem Cells ; cytology ; metabolism ; Octamer Transcription Factor-3 ; metabolism ; Primary Cell Culture ; Retinal Dehydrogenase ; metabolism