Multiple DNases were identified from Haemonchus contortus intestine based on previous studies. The DNases detected at 34, 36 and 38.5 kDa had diverse characteristics. Some of them had characteristics similar to those of mammalians and others had unusual characteristics. This study was carried out to fractionate worm intestinal DNases from other proteins using phenyl Sepharose chromatographic methods. All DNases detected from Haemonchus contortus intestine were fractionated in the flowthrough of phenyl Sepharose, indicating the worm DNases are hydrophilic. The DNases were enriched five-fold in the flowthrough fraction while additional steps are required for isolation of the worm DNases. Thus, fractionation with phenyl Sepharose could be used as a good initial step to enrich and separate DNases from other proteins.
Chromatography ; Deoxyribonucleases ; Haemonchus ; Intestines ; Proteins ; Sepharose

Multiple DNases were identified from Haemonchus contortus intestine based on previous studies. The DNases detected at 34, 36 and 38.5 kDa had diverse characteristics. Some of them had characteristics similar to those of mammalians and others had unusual characteristics. This study was carried out to fractionate worm intestinal DNases from other proteins using phenyl Sepharose chromatographic methods. All DNases detected from Haemonchus contortus intestine were fractionated in the flowthrough of phenyl Sepharose, indicating the worm DNases are hydrophilic. The DNases were enriched five-fold in the flowthrough fraction while additional steps are required for isolation of the worm DNases. Thus, fractionation with phenyl Sepharose could be used as a good initial step to enrich and separate DNases from other proteins.
Chromatography ; Deoxyribonucleases ; Haemonchus ; Intestines ; Proteins ; Sepharose

Trypsin and DNase digestion technique has become a retinal digestion technique for studying diabetic retinopathy. We tried osmotic digestion method with DNase and compared the quality of preparation of retina and microvascular change with trypsin digestion in normal and diabetic rats. Streptozotocin-induced diabetic rats were sacrificed at 9, 18, 27weeks in 6 rats. Right retinas were digested with 3% trypsin while left wer digested with 0.1% DNase. For control, 18 normal rats were sacrificed at the same time. DNase was superior to trypsin for retinal preparation on the stainability, the degree of separating nonvascular fissue from vascular net, the degree of preservation of vascular net in normal & diabetic rats. In diabetic rats, the number of pericyte decreased significantly with age, which not in normal rats(p=0.0023) We suggest DNase digestion technique as an new alternative for trypsin digestion technique in the study of microvascular change of diabetic retinopathy.
Animals ; Deoxyribonucleases* ; Diabetes Mellitus ; Diabetic Retinopathy ; Digestion* ; Endothelial Cells ; Microvessels* ; Pericytes ; Rats* ; Retina ; Retinaldehyde* ; Trypsin*

BACKGROUND: Staphylococcus aureus is most frequently isolated Gram-positive cocci from clinical specimens. The accurate identification of S. aureus is required. Lipovitellin-salt-mannitol (LSM) agar is medium for differentiating S. aureus and coagulase-negative staphylococci (CNS) by mannitol acidification and lipovitellin lipase reaction. In this study, we compare LSM agar with the other conventional methods for differentiating S. aureus and CNS in clinical laboratories, coagulase test, mannitol-salt agar (MSA), and DNase agar. METHODS: One hundred and forty-five isolates of staphylococci from clinical specimens are used. S. aureus and CNS were identified by coagulase test, MSA, DNase agar, and LSM agar. RESULTS: Eighty-nine isolates were identified as S. aureus and 59 isolates were revealed CNS. Compared ability of methods to differntiate S. aureus from CNS, sensitivity and specificity were 98.7% and 96.6% with LSM agar, 92.1% and 96.6% with coagulase test, 96.6% and 93.2% with MSA, 93.3% and 98.3% with DNase agar, respectively. CONCLUSIONS: LSM agar show good discrimination between S. aureus and CNS. LSM agar may be used for identification of S. aureus in clinical laboratories.
Agar* ; Coagulase ; Deoxyribonucleases ; Discrimination (Psychology) ; Gram-Positive Cocci ; Lipase ; Mannitol ; Sensitivity and Specificity ; Staphylococcus aureus* ; Staphylococcus*

The existence of NMDA receptor and a new organizer protein, Shank, in the postsynaptic density was studied with the cultured hippocampal neurons using by double immunofluorescence method. The hippocampi from embryonic 18 days were dissected and hippocampal neurons were obtained from dissociated hippocampi with 0.25% trypsin and 0.1% DNase in PBS. The hippocampal neurons were plated with density 3,600/cm2 on the poly-L-lysine coated coverglass and cultured 37degrees C, 5% CO2 incubator for 5 weeks. The N2 supplemented MEM was used as a culture medium. Following results are obtained from experiments: 1. The 3~5 minor processes from the cell body of hippocampal neurons were observed at 20 hr in vitro. One of the minor processes was elongated and looked like an axon, and another minor processes showed dendritic branching pattern with slender in thickness. 2. The excitatory NMDA receptor colocalized with PSD-95 which is the postsynaptic density protein. The presynaptic protein, synapsin 1, was closely apposed with PSD-95. 3. Shank which is an organizer protein colocalize with NMDA receptor/PSD-95 complex in the postsynaptic density. Shank proteins may be concerned with the cluster formation of NMDA receptor/PSD-95 in the postsynaptic membrane.
Axons ; Deoxyribonucleases ; Fluorescent Antibody Technique ; Immunohistochemistry ; Incubators ; Membranes ; N-Methylaspartate* ; Neurons* ; Post-Synaptic Density ; Trypsin

Zinc finger nuclease (ZFN), which is a chimeric fusion structure between a Cys2-His2 zinc-finger protein (ZFP) and the cleavage domain of Fok I endonuclease, can be used to introduce targeted double-stranded breaks (DSBs). ZFN-mediated cleavage leads to mutations when double-stranded breaks are repaired by homologous recombination (HR) or nonhomologous end joining (NHEJ). In recent years, ZFNs are widely used in the fields of genetic research. In this review, the methodology and technical advantages of ZFNs were briefly discussed.
Animals ; Deoxyribonucleases, Type II Site-Specific ; chemistry ; genetics ; metabolism ; Humans ; Zinc Fingers

Archaea ; enzymology ; genetics ; Archaeal Proteins ; chemistry ; genetics ; Deoxyribonucleases ; chemistry ; genetics ; Gene Editing ; methods

The glucose nonfermenting gram-negative bacilli encountered about 10% of all gram-negative bacilli isolated from clinical material. Therefore, a rapid and correct identification of glucose nonfermenting gram-negative bacilli is impotent for a better management of infectious disease. There are many conventional systems for the identification of glucose nonfermenting gram-negative bacilli but most of them have problems and difficulties. Commercial Kit Systems exist and they are too expensive for daily use in Korea because of high cost. Based on 12 selected tests we propose a new code system, MCRCODE-N for rapid and inexpensive identification of glucose nonfermenting gram-negative bacilli. The selective 12 tests are oxidase, glucose oxidation motihty, urease, DNase arginine dehydrolase, nitrate reduction, gelatin Liquefaction, esculin hydrolysis, mannitol oxidation, maltose oxidation, Lactose oxidation. The 12 tests are divided 4 group and then each group has 3 tests. The result of each group is expressed by the number as below. The positive test is given by specific number (1st test=1, 2nd test=2, 3rd test=4), while any negative result is 0. Each 3 numbers of one group are added and make number of 1 digit. Four digit number is referred to the code book of MCRCODE-N system or MCRCODE system using computer (Apple-II model) created by authors. This MCRCODE-N system is suitable ones for out use in Korea. We propose the MCRCODE-N system for clinical use.
Arginine ; Clinical Coding* ; Communicable Diseases ; Deoxyribonucleases ; Esculin ; Gelatin ; Glucose Oxidase ; Glucose* ; Hydrolysis ; Korea ; Lactose ; Maltose ; Mannitol ; Urease

OBJECTIVETo construct a reverse transcriptase-polymerase chain reaction (RT-PCR)approach that can improve the specificity of primers while dropping down the nonspecific amplification.

METHODSIn the recent study we reported a new RT-PCR assay which improved markedly the specificity. However its efficiency of regressing nonspecific amplification remains to be accurately checked and further documented. In primer design, we looked over again some sequences that showed differences at 5' or 3' ends between human CAV1 and mouse Cav1 genes. cDNAs and the diluted plasmids which harbored the sequence of human CAV1 or mouse Cav1 gene were chosen as the templates. The ordinary PCR compared with one, of which primers modified by phosphorothioate and combined with proofreading polymerase, for their efficiencies of nonspecific amplification inhibited.

RESULTSTaq DNA polymerase without proofreading activity could efficiently catalyze the extension of primers with a single or multiple mismatched base pairs at the 3' terminus, but the kind of primer extension can be effectively blocked by phosphorothioate modified primers combined with proofreading polymerase. Compared with ordinary PCR reaction, this new PCR method can effectively regress the primer mismatched amplification of 50 ng DNA almost equaling to 2 x 10(4) unmatched template copies in a final volume of 50 microL.

CONCLUSIONCompared with the first generation of polymerases with or without proofreading activities mediating RT-PCR reaction, the introduction of nuclease-resistant 3' modified primers (3' phosphorothioate primer extension) can offer more simplicity, accuracy, and also decrease cost.

Animals ; Caveolin 1 ; genetics ; Deoxyribonucleases ; metabolism ; Gene Amplification ; Humans ; Mice ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Thionucleotides ; metabolism

Cedecea davisae is a motile, Gram-negative rod in the family Enterobacteriaceae which is positive for lipase, DNase and catalase, and negative for gelatinase and oxidase. This bacterium is rarely isolated in the clinical specimens. We isolated C. davisae from the ascitic fluid of a 49-year old male patient with liver cirrhosis who was diagnosed as acute bacterial peritonitis. Bacterial identification was performed by API 20E and VITEK. Antimicrobial susceptibility test showed that the isolate was susceptible to cefotaxime, piperacillin, and imipenem. Peritonitis of this patient was improved by imipenem therapy. This is the first reported case of peritonitis caused by this organism.
Ascitic Fluid ; Catalase ; Cefotaxime ; Deoxyribonucleases ; Enterobacteriaceae ; Gelatinases ; Humans ; Imipenem ; Lipase ; Liver Cirrhosis* ; Liver* ; Male ; Middle Aged ; Oxidoreductases ; Peritonitis* ; Piperacillin