The present study was undertaken to observe the quality and quantity of lipid constituents by the developing Ascaris suum eggs. The collected eggs from the uterus of A. suum were classified into 3 groups, i.e., single cell stage, morula stage and embryonated eggs, and were subjected to analyse their lipid fractions. To obtain the morula stage eggs, 10 to 11 incubation days at 30 degree C were needed and for the embryonated eggs, 30 to 31 days were lasted. At the time of experiment, their indices of development by Hoffman were 285(morula stage) and 42 (embryonated stage) respectively. Lipid extraction was done by the methods of Folch et a1. (1957) and Kenny (1952), and then the extracted lipid fractions from the above 3 groups of eggs were separated by thin layer chromatography. Those fractions were also subjected to perform the quantitative analyses of fatty acids, glycerides, cholesterol and phospholipids. The results obtained were summarized as follows. Total amount of fatty acid was decreased from 12.9 mg per gram of eggs (single cell stage) to 6.6 mg/gm (embryonated eggs), whereas the proportion of free fatty acid to total fatty acid was constantly increased from 77.5 percent to 89.4 percent during the period of egg development. Total amount of glycerides was also increased from 33.0 mg/gm of single cell stage to 55.9 mg/gm of the embryonated eggs. The most abundant glyceride among 3 glycerides discovered from A. suum eggs was triglyceride, and the least was monoglyceride. The amount of free cholesterol was much larger than that of ester form in general, and it reached maximum in the eggs of morula stage (4.6 mg/gm). The increase of total cholesterol was monitored during the development of A. suum eggs from 3.3 mg/gm to 5.4 mg/gm. The following 8 phospholipids were detected in the embryonated eggs, i.e., lysophosphatidyl choline, phosphatidyl inositol, sphingomyelin, phosphatidyl choline, phosphatidyl glycerol, phosphatidyl serine, phosphatidyl ethanolamine and unknown phospholipid. But in the single cell stage eggs, 4 kinds out of the above 8 phopholipids were not observed, and in the morula stage eggs, 2 kinds were absent among the 8 phospholipids.
parasitology-helminth-nematoda ; Ascaris suum ; ovum ; lipid ; fatty acid ; glycerides ; triglyceride ; monoglyceride ; cholesterol ; lysophosphatidyl choline ; phosphatidyl inositol ; sphingomyelin ; phosphatidyl choline ; phosphatidyl glycerol ; phosphatidyl serine ; phosphatidyl ethanolamine ; phopholipids ; biochemistry

BACKGROUND: Paroxysmal nocturnal hemoglobinuria (PNH) is caused by deficient biosynthesis of the glycosylphosphatidylinositol (GPI) anchor in hemopoietic stem cells. Mutation of phosphatidyl inositol glycan class A (PIG-A) gene, an X-linked gene that participates in the first step of GPI anchor biosynthesis, is responsible for PNH. Characteristics of somatic mutation of PIG-A gene in the Korean patients with PNH and their relationships to clinical characteristics were analyzed. METHODS: Twenty five patients with PNH and a donor of bone marrow transplantation were selected. Ham tests, sucrose hemolysis tests and CD59 expressions of erythrocytes and granulocytes were performed to confirm diagnosis. Dideoxy fingerprinting (ddF) was used to screen mutations, and direct sequencing of DNA was performed to characterize the mutations. RESULTS: The mutations of PIG-A gene were found in twelve cases and ten of them were novel mutations. There were five deletions, six substitutions and a insertion. Therewere six premature terminations, three abnormal splicings, a missense and two nonsense mutations. There were six point mutations and six frameshift mutations. Five cases of hypoplastic PNH showed mutations only in exons, but three in seven cases of hemolytic PNH showed mutations in introns. Two cases with symptoms of venous thrombosis showed mutations in exon 3. CONCLUSION: There were ten novel mutations among twelve mutations in the Korean patients with PNH and characteristics of the mutations were variable without any remarkable hot spot in sites and types. The characteristics of mutation were not correlated with the results of clinical courses of the patients with PNH.
Bone Marrow Transplantation ; Codon, Nonsense ; Dermatoglyphics ; Diagnosis ; DNA ; Erythrocytes ; Exons ; Frameshift Mutation ; Genes, X-Linked ; Glycosylphosphatidylinositols ; Granulocytes ; Hemoglobinuria, Paroxysmal* ; Hemolysis ; Humans ; Introns ; Phosphatidylinositols ; Point Mutation ; Stem Cells ; Sucrose ; Tissue Donors ; Venous Thrombosis

No abstract available.
Immunoglobulins, Thyroid-Stimulating* ; Phosphatidylinositols* ; Thyroid Gland*

PURPOSE: To investigate the effects of nitroglycerin on the production of nitric oxide and its related pathway in cultured human trabecular meshwork cells (HTMC). METHODS: Primarily cultured HTMC were exposed to 10 nM nitroglycerin using 1% serum-containing media for 30 minutes. The production of nitric oxide was assessed with the Griess assay and expressions of eNOS mRNA was assessed with RT-PCR. Additionally, the cells were exposed to wortmanin and Akt1/2 kinase inhibitor to investigate the mechanism related to the production of nitric oxide. RESULTS: Nitroglycerin increased the production of nitric oxide (p < 0.05) accompanied with increased expression of eNOS mRNA. The increased production of nitric oxide and eNOS mRNA was inhibited by wortmanin and Akt1/2 kinase inhibitor. CONCLUSIONS: Low-dose nitroglycerin increased the production of nitric oxide accompanied by increased eNOS activity. Nitroglycerin drives eNOS activation via the phosphatidylinositol 3-kinase/protein kinase B pathway.
Humans ; Nitric Oxide ; Nitroglycerin ; Phosphatidylinositols ; Phosphotransferases ; RNA, Messenger ; Trabecular Meshwork

Sinonasal mucosal melanoma (SMM) is an aggressive and rare type of melanoma. Although the classic RAS-RAF-MEK pathway is thought to be the main pathway involved in melanoma pathogenesis, genetic alterations in the phosphatidylinositol 3-kinase-AKT pathway, including PTEN-regulated signaling, are also thought to contribute. So far, data regarding altered PTEN expression and epigenetic mechanism of PTEN silencing in development of SMM is extremely limited. Herein we report on a case of SMM with liver and bone metastases with an epigenetic alteration of PTEN. Results of mutation analysis for BRAF, NRAS, HRAS, KRAS, PIK3CA, c-Kit, and PTEN were negative; however, methylation of PTEN CpG islands was observed. Our case not only supports PTEN as a major tumor suppressor involved in melanoma tumorigenesis, but also a potential epigenetic mechanism of PTEN silencing in development of SMM.
Carcinogenesis ; CpG Islands ; Epigenomics ; Liver ; Melanoma* ; Methylation* ; Neoplasm Metastasis ; Phosphatidylinositols

Sinonasal mucosal melanoma (SMM) is an aggressive and rare type of melanoma. Although the classic RAS-RAF-MEK pathway is thought to be the main pathway involved in melanoma pathogenesis, genetic alterations in the phosphatidylinositol 3-kinase-AKT pathway, including PTEN-regulated signaling, are also thought to contribute. So far, data regarding altered PTEN expression and epigenetic mechanism of PTEN silencing in development of SMM is extremely limited. Herein we report on a case of SMM with liver and bone metastases with an epigenetic alteration of PTEN. Results of mutation analysis for BRAF, NRAS, HRAS, KRAS, PIK3CA, c-Kit, and PTEN were negative; however, methylation of PTEN CpG islands was observed. Our case not only supports PTEN as a major tumor suppressor involved in melanoma tumorigenesis, but also a potential epigenetic mechanism of PTEN silencing in development of SMM.
Carcinogenesis ; CpG Islands ; Epigenomics ; Liver ; Melanoma* ; Methylation* ; Neoplasm Metastasis ; Phosphatidylinositols

The ovarian follicles develop initially from primordial follicles. The majority of ovarian primordial follicles are maintained quiescently as a reserve for the reproductive life span. Only a few of them are activated and develop to an advanced follicular stage. The maintenance of dormancy and activation of primordial follicles are controlled by coordinated actions of a suppressor/activator with close communications with somatic cells and intra-oocyte signaling pathways. Many growth factors and signaling pathways have been identified and the transforming growth factor-beta superfamily plays important roles in early folliculogenesis. However, the mechanism of maintaining the dormancy and survival of primordial follicles has remained unknown for decades. Recently, since the first finding that all primordial follicles are activated prematurely in mice deficient forkhead box O3a, phosphatidylinositol 3 kinase/phosphatase and tensin homolog (PTEN) signaling pathway was reported to be important in the regulation of dormancy and initial follicular activation. With these informations on early folliculogenesis, clinical application can be expected such as in vitro maturation of immature oocytes or in vitro activation of follicles by PTEN inhibitor in cryopreserved ovarian cortical tissues for fertility preservation.
Animals ; Female ; Fertility Preservation ; Intercellular Signaling Peptides and Proteins ; Mice ; Microfilament Proteins ; Oocytes ; Ovarian Follicle ; Phosphatidylinositols

The ovarian follicles develop initially from primordial follicles. The majority of ovarian primordial follicles are maintained quiescently as a reserve for the reproductive life span. Only a few of them are activated and develop to an advanced follicular stage. The maintenance of dormancy and activation of primordial follicles are controlled by coordinated actions of a suppressor/activator with close communications with somatic cells and intra-oocyte signaling pathways. Many growth factors and signaling pathways have been identified and the transforming growth factor-beta superfamily plays important roles in early folliculogenesis. However, the mechanism of maintaining the dormancy and survival of primordial follicles has remained unknown for decades. Recently, since the first finding that all primordial follicles are activated prematurely in mice deficient forkhead box O3a, phosphatidylinositol 3 kinase/phosphatase and tensin homolog (PTEN) signaling pathway was reported to be important in the regulation of dormancy and initial follicular activation. With these informations on early folliculogenesis, clinical application can be expected such as in vitro maturation of immature oocytes or in vitro activation of follicles by PTEN inhibitor in cryopreserved ovarian cortical tissues for fertility preservation.
Animals ; Female ; Fertility Preservation ; Intercellular Signaling Peptides and Proteins ; Mice ; Microfilament Proteins ; Oocytes ; Ovarian Follicle ; Phosphatidylinositols

Sorting nexins (SNXs) are a large group of proteins that contain Phox (PX) domain and involve in regulating endocytosis and endosome sorting. SNX7, a member of SNXs family, contains a PX domain and a BAR domain. In zebrafish, SNX7 is a liver-enriched anti-apoptotic protein and indispensible for the liver development. A fragment of SNX7 cDNA ((px-bar)snx7), encoding the PX domain and the BAR domain, was inserted into the expressing vector p28a, transformed into E. coli Rosseta 2 (DE3), and then induced by isopropyl β-D-1-Thiogalactopyranoside (IPTG). After affinity, ion exchange and gel filtration purification, the purity of (PX-BAR)SNX7 reached over 95%. Dynamic light scattering (DLS) experiment indicated that (PX-BAR)SNX7 was homogeneous in solution. Lipid overlay assay showed that (PX-BAR)SNX7 can bind to PtdIns(5)P, PtdIns(4,5)P2 and PtdIns(3,4,5)P3.
Escherichia coli ; metabolism ; Genetic Vectors ; Humans ; Phosphatidylinositols ; metabolism ; Recombinant Proteins ; biosynthesis ; Sorting Nexins ; biosynthesis ; Substrate Specificity

OBJECTIVE: To investigate the effect of electro-acupuncture (EA) on vasomotor symptoms in rats with acute cerebral infarction, by observing the changes in the expression of factors related to the phosphatidylinositol (PI) system. METHODS: Forty-two Wistar rats were randomly divided into 3 groups by a random number table: the control group (n=6), the model group (n=18) and the EA group (n=18). The EA group was given EA treatment at Shuigou (GV 26) instantly after modeling with middle cerebral artery occlusion (MCAO) method, while the model and control groups were not given any treatment. The degrees of neurological deficiency were evaluated using neurological severity scores (NSS) and the brain blood flow was evaluated by a laser scanning confocal microscope. Western blot analysis was conducted to detect the expression levels of G-protein subtype (Gq) and calmodulin (CaM). Competition for protein binding was conducted to detect the expression level of inositol triphosphate (IP3). Thin layer quantitative analysis was conducted to detect the expression level of diacylglycerol (DAG). The expression level of intracellular concentration of free calcium ion ([Ca RESULTS: The NSS of the model group was significantly higher than the control group at 3 and 6 h after MCAO (P<0.01), while the EA group was significantly lower than the model group at 6 h (P<0.01). The cerebral blood flow in the model group was significantly lower than the control group at 1, 3 and 6 h after MCAO (P<0.01), while for the EA group it was remarkably higher than the model group at the same time points (P<0.01). The expressions of Gq, CaM, IP3, DAG and [Ca CONCLUSION EA treatment at GV 26 can effectively decrease the over-expression of related factors of PI system in rats with acute cerebral infarction, improve cerebral autonomy movement, and alleviate cerebral vascular spasm.
Acupuncture Therapy ; Animals ; Brain Ischemia ; Cerebral Infarction/therapy* ; Electroacupuncture ; Phosphatidylinositols ; Rats ; Rats, Wistar