Surface ultrastructure of Parvatrema timondavidi developmental stages was studied using a scanning electron microscope. The metacercariae were collected from the marine clam, Tapes philippinarum, and juvenile and worms adult were recovered at 1, 2, 3, and 7 days after experimental infection of mice. The metacercariae had a large oral sucker and characteristic lateral projections. Around the lip of the oral sucker many type I and type II sensory papillae were observed, and type III papillae were located symmetrically on the medial side of the lateral projection. Numerous type I papillae were grouped around the genital pore. The tegumental spines were distributed over the worm surface except the lip of the sucker and genital pore. The 1-day old worm had a well-developed ventral sucker, with 6 type II sensory papillae on its outer surface and another 6 type I papillae on the inner side, Two small type I papillae were seen on the anterior side of the ventral sucker. The genital pore was and 15 type I papillae were grouped around it. The 2-, 3-, and 7-day worms revealed that as they grew to be adults, the spine tips became multipointed, the genital pore formed a genital atrium, and the cytoplasmic process became well differentiated. In 2- and 3-day worms 10 type II papillae encircling the lip of the oral sucker, and additional 4 papilled at the dorsal side of 4 dorsal type II papillae were a characteristic feature. The distribution pattern of sensory papillae around the oral sucker and genital pore, and 2 type I papillae on the anterior side of the ventra sucker, was so peculiar in P. timondavidi, that they seem to be useful keys for taxonomic differentiation from other gymnophallids.
parasitology-helminth-trematoda ; Parvatrema timondavidi ; surface ultrastructure ; scanning EM, sensory papilla ; spine ; cytoplasmic process

Neural prosthesis control system is based on brain-computer interface and functional electrical stimulation technology, by an-alyzing the electroencephalograph control commands directly into the muscle system or an external device, which compensated efferent pathway from the brain-spinal cord, and recovered motor function of patients with cervical spinal cord injury. This paper described the basic structure, working principle and key technology of neural prosthetic system, summarized the application, problems and prospects of neural prosthetic technology in the rehabilitation of cervical spinal cord injury.

OBJECTIVE:To optimize the decoction process of agkistrodon. METHODS:RP-HPLC with pre-column derivatiza-tion was adopted. With the contents of 4 main amino acids in agkistrodon as index,the decoction process(decocting times,heating time,water quantity and medicinal material crushing granularity) was optimized by orthogonal tests and verified. RESULTS:The optimal technology of the decoction process of agkistrodon was as 3 times of decoction,60 min of heating time,50 ml of water consumption for 0.90 g medicinal material and No.6 sieve. The results of verification test showed the total extraction of 4 kinds of amino acids was 72.68 mg/g(RSD=3.77%,n=3). CONCLUSIONS:The decoction technology can be used for the decoction pro-cess of agkistrodon,and it is stable and feasible.

Cryptosporidium, a coccidian parasite first described by Tyzzer (1907) from a laboratory mouse, has become an important human enteric pathogen causing overwhelming diarrhea especially in immunocompromised patients such as AIDS. This parasite has been reported from over 20 countries and is recognized as a cosmopolitan species. In Korea, however, there has been no report on human as well as animal cryptosporidiosis. This study was performed so as to verify the presence of Cryptosporidium in Korea by activating the parasite from laboratory mice by immunosuppression. Total 65 conventionally-bred ICR mice including a control (5 mice) and 3 experimental groups (20 each) were used for this study. Group I was immunosuppressed with prednisolone injection (1 mg IM, every other day) for 7 weeks. Group II (prednisolone injection and tetracycline administration) and Group III (prednisolone injection and trimethoprim-sulfamethoxazole administration) were prepared to observe the effect of antibacterial agents on the activation of cryptosporidiosis. In fecal examinations of mice Cryptosporidium oocysts (4-6 microns in size) were detected from 1 week after the start of immunosuppression and the mice began to die. In H-E stained tissue sections of the lower jejunum, numerous very small (2-4 microns), dense, ovoid or spherical, slightly basophilic bodies were seen attached on the free border of mucosal epithelial cells. In scanning and transmission electron microscopic observations, these organisms were identified as various developmental stages of Cryptosporidium. The species is considered to be C. parvum.(ABSTRACT TRUNCATED AT 250 WORDS)
Cryptosporidiosis-etiology ; Cryptosporidiosis-immunology ; Cryptosporidium-growth-and-development ; English-Abstract ; Immune-Tolerance ; Intestinal-Diseases,-Parasitic-etiology ; Intestinal-Diseases,-Parasitic-immunology ; Mice- ; Mice,-Inbred-ICR ; *Cryptosporidiosis-parasitology ; *Cryptosporidium-pathogenicity ; *Immunosuppression- ; *Intestinal-Diseases,-Parasitic-parasitology

We investigated the optimal culture conditions for Cryptosporidium muris in a human stomach adenocarcinoma (AGS) cell line by determining the effects of medium pH and of selected supplements on the development of C. muris. The optimum pH of the culture medium required for the development of C. muris was determined to be 6.6. The number of parasites significantly increased during cultivation for 72 hr (p < 0.05) at this level. On the other hand, numbers decreased linearly after 24 hr of incubation at pH 7.5. When cultured in different concentrations of serum, C. muris in media containing 5% FBS induced 4-7 times more parasites than in 1% or 10% serum. Of the six medium supplements examined, only 1 mM pyruvate enhanced the number of C. muris in vitro. Transmission electron microscopic observation showed the developmental stages of C. muris in the cytoplasm of the cells, not in an extracytoplasmic location. The growth of C. muris in AGS cells provides a means of investigating its biological characteristics and of testing its response to therapeutic agents. However, a more optimized culture system is needed for the recovery of oocysts on a large scale in vitro.
Adenocarcinoma ; Animals ; Cell Line, Tumor ; Cryptosporidium/*growth & development ; Culture Media ; Human ; Hydrogen-Ion Concentration ; Stomach/*parasitology ; Stomach Neoplasms ; Support, Non-U.S. Gov't

BACKGROUND/AIMS: The introduction of Hansenula polymorpha for recombinant hepatitis B vaccine production allowed high product yield with plasmid stability and less glycosylation than conventional Saccharomyces cerevisiae system. A Green Cross HG-II vaccine formulated from HBsAg produced by a recombinant strain of the yeast H. polymorpha was evaluated for immunogenicity and safety in an open label triaL METHOFD: A 20 ug dose of Green Cross HG-II vaccine was administered intramuscularly at 0, 1 and 6 months at the deltoid region in 118 healthy adults seronegative for HBV markers. The anti-HBs titers were determined at one month after administration of the third dose of vaccine by radioimmunoassay. RESULTS: The seroconversion rate was 96.8% (90 out of 93), with seroprotective rate of 95.7% (89 out of 93). The geometric mean titers(GMT) of the anti-HBs response was 153.1mIU/ml in seroconverters. An age-dependent effect was observed in the anti-HBs response. But sex-dependent effect was not prominent. Reactogenecity was in incidence and general reactions were short-lasting and a mainly mild in severity. CONCLUSIONS: The results of this study have shown that the Green Cross HG-II vaccine is safe and clinically well tolerated, a nd that it may provide protection against HBV infection.
Adult* ; Glycosylation ; Hepatitis B Surface Antigens ; Hepatitis B Vaccines* ; Hepatitis B* ; Hepatitis* ; Humans ; Incidence ; Pichia ; Plasmids ; Radioimmunoassay ; Saccharomyces cerevisiae ; Yeasts

The present study was undertaken to know the infection status of Cryptosporidium parvum among the residents of Chorwon-gun, Kangwon-do in 1993. Total 461 fecal samples were collected from the inhabitants residing in Chorwon-gun during the period of August 12 to September 14, 1993. Fecal smears were prepared by formalin-ether sedimentation, and examined after modified acid fast staining. Of the 461 fecal samples, 9 (1.9%) were positive for C. parvum oocysts. The positive cases were limited to thirties (4) patients, forties (3), and sixties (2), and no oocyst was detected in other age groups. The oocyst positive rate for male was 1.4% and that of female was 2.6%.
Adolescent ; Adult ; Age Factors ; Aged ; Animals ; Child ; Cryptosporidiosis/*epidemiology/parasitology ; Cryptosporidium parvum/isolation & purification ; Feces/parasitology ; Female ; Human ; Korea/epidemiology ; Male ; Middle Aged ; Parasite Egg Count ; Sex Factors

OBJECTIVES: Many studies have suggested different neurobiological findings and clinical courses in alcoholism. Recently, subtyping in alcohol dependence has become essential to overcome the heterogeneity of patients. Among several criteria of subtypes, Lesch's typology is proposed to integrate biological, social, and psychological factors. This review provides neurobiological findings and treatment-responses of alcohol dependence according to Lesch's typology. METHOD: We searched the international published medical literature using the search terms 'Lesch's typology' and 'alcohol dependence' and using the limits 'human'. RESULTS: We identified 17 studies with subjects of alcohol dependence according to Lesch's typology. CONCLUSION: They indicated that each subtype of Lesch's typology can have specific neurobiological factors and different clinical responses as follows. Lesch's subtype 1 is characterized by severe withdrawal symptoms and associated with elevated glutamate and homocysteine. Lesch's subtype 2 is defined by individuals who drink alcohol as self-medication for anxiety. Their craving has significant positive correlations with prolactin, leptin level, or intake-volume (vasopressin). Lesch's subtype 4 is related to cerebral dysfunction and associated with increased glutamate and left-handedness. Clinical trials showed that naltrexone was effective in Lesch's subtype 3 and 4 patients, while acamprosate was effective in the subtypes 1 and 2.
Alcoholism ; Anxiety ; Glutamic Acid ; Homocysteine ; Humans ; Leptin ; Naltrexone ; Population Characteristics ; Prolactin ; Substance Withdrawal Syndrome ; Taurine

AIMTo investigate the activation of the nuclear factor of activated T cells (NFAT) and its function in the corticosterone (CORT)-induced apoptosis of rat Leydig cells.

METHODSNFAT in rat Leydig cells was detected by Western blotting and immunohistochemical staining. Cyclosporin A (CsA) was used to evaluate potential involvement of NFAT in the CORT-induced apoptosis of Leydig cells. Intracellular Ca(2+) was monitored in CORT-treated Leydig cells using Fluo-3/AM. After the Leydig cells were incubated with either CORT or CORT plus CsA for 12 h, the levels of NFAT2 in the nuclei and in the cytoplasm were measured by semi-quantitative Western blotting. The role of NFAT2 in CORT-induced Leydig cell apoptosis was further evaluated by observing the effects of NFAT2 overexpression and the inhibition of NFAT2 activation by CsA on FasL expression and apoptosis.

RESULTSWe found that NFAT2 was the predominant isoform in Leydig cells. CsA blocked the CORT-induced apoptosis of the Leydig cells. The intracellular Ca(2+) level in the Leydig cells was significantly increased after the CORT treatment. The CORT increased the level of NFAT2 in the nuclei and decreased its level in the cytoplasm. CsA blocked the CORT-induced nuclear translocation of NFAT2 in the Leydig cells. Both CORT-induced apoptosis and FasL expression in the rat Leydig cells were enhanced by the overexpression of NFAT2 and antagonized by CsA.

CONCLUSIONNFAT2 was activated in CORT-induced Leydig cell apoptosis. The effects of NFAT2 overexpression and the inhibition of NFAT2 activation suggest that NFAT2 may potentially play a pro-apoptotic role in CORT-induced Leydig cell apoptosis through the up-regulation of FasL.

Animals ; Apoptosis ; drug effects ; Calcium ; metabolism ; Cell Nucleus ; metabolism ; Corticosterone ; pharmacology ; Cytoplasm ; metabolism ; Immunohistochemistry ; Kinetics ; Leydig Cells ; cytology ; drug effects ; physiology ; Male ; NFATC Transcription Factors ; metabolism ; Rats ; Rats, Sprague-Dawley

Human bone marrow mesenchymal stem cells (MSCs) expanded in vitro exhibit not only a tendency to lose their proliferative potential, homing ability and telomere length but also genetic or epigenetic modifications, resulting in senescence. We compared differential methylation patterns of genes and miRNAs between early-passage [passage 5 (P5)] and late-passage (P15) cells and estimated the relationship between senescence and DNA methylation patterns. When we examined hypermethylated genes (methylation peak > or = 2) at P5 or P15, 2,739 genes, including those related to fructose and mannose metabolism and calcium signaling pathways, and 2,587 genes, including those related to DNA replication, cell cycle and the PPAR signaling pathway, were hypermethylated at P5 and P15, respectively. There was common hypermethylation of 1,205 genes at both P5 and P15. In addition, genes that were hypermethylated at P5 (CPEB1, GMPPA, CDKN1A, TBX2, SMAD9 and MCM2) showed lower mRNA expression than did those hypermethylated at P15, whereas genes that were hypermethylated at P15 (MAML2, FEN1 and CDK4) showed lower mRNA expression than did those that were hypermethylated at P5, demonstrating that hypermethylation at DNA promoter regions inhibited gene expression and that hypomethylation increased gene expression. In the case of hypermethylation on miRNA, 27 miRNAs were hypermethylated at P5, whereas 44 miRNAs were hypermethylated at P15. These results show that hypermethylation increases at genes related to DNA replication, cell cycle and adipogenic differentiation due to long-term culture, which may in part affect MSC senescence.
Bone Marrow Cells/*metabolism ; *DNA Methylation ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; Mesenchymal Stromal Cells/*metabolism ; MicroRNAs ; Molecular Sequence Annotation ; Primary Cell Culture ; Promoter Regions, Genetic ; Reproducibility of Results ; Signal Transduction ; Telomere Shortening