Excretory/secretory proteins (ESP) from Toxoplasma gondii were analyzed to define the function in the penetration process into host cells. Whole ESP obtained at 37 degrees C were composed of 15 bands with molecular mass of 110, 97, 86, 80, 70, 60, 54, 42, 40, 36, 30, 28, 26, 22, and 19 kDa. Five ESP of 86, 80, 42, 36, and 28 kDa were reacted with monoclonal antibodies (mAb), named as Tg386 (microneme), Tg485 (surface membrane), Tg786 (rhoptry), Tg378, and Tg556 (both dense granules), respectively. The ESP was released by a temperature-dependent/-independent manner and all at once whenever ready to pour out except Tg786. Each ESP was not exhausted within the parasite but the amount was limited. Tg786 was released continuously with increment, whereas Tg378 and Tg556 were ceased to release after 3 and 4 hr. Dense granular Tg378 and Tg556 were released spontaneously and constitutively before the entry into host cells also. The entry of T. gondii was inhibited by all the mAbs differentially. And the parasite deprived of ESP was inhibited to enter exponentially up to 90.1%. It is suggested that ESP play an essential function to provide appropriate environment for the entry of the parasite into host cells.
Animals ; *Antibodies, Monoclonal ; *Antibodies, Protozoan ; Antigens, Protozoan/*analysis/physiology ; Mice ; Mice, Inbred BALB C ; Support, Non-U.S. Gov't ; Temperature ; Toxoplasma/*chemistry/pathogenicity

The antigenic domain of the major surface protein (Nc-p43) of Neospora caninum was examined by polymerase chain reaction of its gene fragments and recombinant expression as GST fusion proteins. The fragments of Nc-p43 were as follow: a total open reading frame (OFR), T; OFR without signal sequence and C-terminal hydrophobic sequence, S; N-terminal 2/3 parts of S, A; C-terminal 2/3 parts, P; N-terminal 1/3 part, X; middle 1/3 part, Y; and C-terminal 1/3 part, Z, respectively. The DNA fragments were cloned into pGEX-4T vector. Recombinant plasmids transformed into Escherichia coli of BL21 pLysS (DE3) strain were induced to express GST or GST fused fragments of Nc-p43 such as 69 kDa protein for T, 66 kDa for S, 52 kDa for A, 53 kDa for P, and 40 kDa proteins for X, Y, and Z, respectively in SDS-PAGE. The Nc-p43 fragments of T, S, and P reacted with a bovine serum of neosporosis while those of A, X, Y, and Z together with GST did not in the western blot. These findings suggest that the antigenic domain of Nc-p43 of N. caninum may be localized in the C-terminal 2/3 parts. Together with A19 clone in SAG1 of Toxoplasma gondii (Nam et al., 1996), the P fragment of Nc-p43 could be used as efficient antigens to diagnose and differentiate those infections with both species.
Amino Acid Sequence ; Animals ; Base Sequence ; Biological Markers/analysis ; Blotting, Western ; Cells, Cultured ; Cercopithecus aethiops ; Coccidiosis/diagnosis ; Neospora/*immunology ; Protozoan Proteins/*analysis/genetics/immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Support, Non-U.S. Gov't ; Vero Cells/parasitology

PURPOSE: To evaluate the value of dacryoscintigraphy in the assessment of patients with a clinical diagnosis of functional nasolacrimal duct obstruction. MATERIALS AND METHODS: Dacryoscintigraphy was performed in symptomatic 35 lacrimal drainage systems in 18 patients (age range: 37~76, 8 males, 10 females) that were patent on syringing. RESULTS: Abnormalities were detected with dacryoscintigraphy in 75.8% of systems. The positive scintigrams were subdivided into those demonstrating prelacrimal sac delay (31.8%), delay at the lacrimal sac/junction (40.9%), or delay within the duct (27%). CONCLUSION: Dacryoscintigraphy is noninvasive useful technique in the assessment of the functional nasolacrimal duct obstruction and very sensitive at detecting abnormalities in patients with lid laxity caused by senile change and facial nerve palsy.
Diagnosis* ; Drainage ; Facial Nerve ; Humans ; Male ; Nasolacrimal Duct* ; Paralysis

Western blot analysis was performed to diagnose vivax malaria using stage-specific recombinant antigens. Genomic DNA from the whole blood of a malaria patient was used as templates to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), merozoite surface protein (MSP-1), apical merozoite antigen (AMA-1), serine repeat antigen (SERA), and exported antigen (EXP-1) of Plasmodium vivax. Each amplified DNA fragment was inserted into a pGEX-4T plasmid to induce the expression of GST fusion protein in Escherichia coli by IPTG. The bacterial cell extracts were separated on 10% SDS-PAGE followed by western blot analysis with patient sera which was confirmed by blood smear examination. When applied with patient sera, 147 (91.9%) out of 160 vivax malaria, 12 (92.3%) out of 13 falciparum malaria, and all 9 vivax/falciparum mixed malaria reacted with at least one antigen, while no reactions occurred with 20 normal uninfected sera. In the case of vivax malaria, CSP-1 reacted with 128 (80.0%) sera, MSP-1 with 102 (63.8%), AMA-1 with 128 (80.0%), SERA with 115 (71.9%), and EXP-1 with 89 (55.6%), respectively. We obtained higher detection rates when using 5 antigens (91.9%) rather than using each antigen solely (55.6-80%), a combination of 2 (76.3-87.5%), 3 (85.6-90.6%), or 4 antigens (89.4-91.3%). This method can be applied to serological diagnosis, mass screening in endemic regions, or safety test in transfusion of prevalent vivax malaria.
Animals ; Antigens, Protozoan/*blood ; Biological Markers/blood ; *Blotting, Western ; Human ; Life Cycle Stages/immunology ; Malaria, Vivax/*diagnosis ; Mass Screening ; Plasmodium vivax/*immunology ; Recombinant Proteins/blood ; Serologic Tests/methods ; Support, Non-U.S. Gov't

A cDNA of 1.1 kb comprising the gene encoding the peroxiredoxin of Toxoplasma gondii (TgPrx) has been cloned. The open reading frame of 591 bp was translated into a protein of 196 amino acids with a molecular mass of 25 kDa. Conserved 2 cysteine domains of Phe-Val-Cys-Pro and Glu-Val-Cys-Pro indicated TgPrx belonged to 2-Cys Prx families. TgPrx showed the highest homology with that of Arabidopsis thaliana by 53.9% followed by Entamoeba histolytica with 39.5% by the amino acid sequence alignment. Polyclonal antibody against recombinant TgPrx detected 25 kDa band in T. gondii without binding to host cell proteins. TgPrx was located in the cytoplasm of T. gondii extracellularly or intracellularly by immunofluorescence assay. The expression of TgPrx was increased as early as 30 min after the treatment with artemisinin in the intracellular stage, while no changes in those of host Prx I and TgSOD. This result implies that TgPrx may function as an antioxidant protecting the cell from the attack of reactive oxygen intermediates. It is also suggested that TgPrx is a possible target of chemotherapy.
Amino Acid Sequence ; Animals ; Antioxidants ; *Artemisinins ; Base Sequence ; *Cloning, Molecular ; Cysteine/metabolism ; Molecular Sequence Data ; Nucleic Acid Amplification Techniques ; Peroxidases/chemistry/*genetics/physiology ; Sesquiterpenes/pharmacology ; Support, Non-U.S. Gov't ; Toxoplasma/*enzymology

PURPOSE: Thallim-201 (201Tl) brain SPECT and proton (1H) magnetic resonance spectroscopy (MRS) have been used to evaluate tumor grade and viability of glioma. We assessed the correlations between 201Tl brain index or spectrum of metabolites of 1H MRS and grade of glioma or histopathologic findings. MATERIALS AND METHODS: We studied 17 patients (4 astrocytoma, 7 anaplastic astrocytoma and 6 glioblastoma). On 201Tl Brain SPECT, 201Tl index was measured as the ratio of average counts for region of interest to those for the contralateral normal brain. On 1H MRS, we calculated choline (Cho) /creatine (Cr) ratio and N-acetylaspartate (NAA)/Cr ratio in ROI defined as tumor center. Histopathologic findings were graded by Ki-67 index, cellularity, mitosis, pleomorphism, necrosis and endothelial proliferation. An unpaired t test and statistical correlations were performed to evaluate these data. RESULTS: Tl-index showed the best correlation with Ki-67 index (p<0.01), less correlations with cellularity, mitosis, and endothelial proliferation, but no correlation with results of MRS, pleomorphism, or necrosis. The findings of MRS did not correlate with all of the above. The cases of glioblastoma demonstrated a higher Tl-index, Cho/Cr ratio, Ki-67 index and lower NAA/Cr ratio, albeit without statistical significance. CONCLUSION: Even though 201Tl brain SPECT did not correlate directly with grade of malignancy, it may still be useful in determining biological aggressiveness of tumor and prognosis of patients because it correlated well with Ki-67 index, a growth fraction of glioma, cellularity, mitosis and endothelial proliferation.
Astrocytoma ; Brain* ; Choline ; Glioblastoma ; Glioma* ; Humans ; Magnetic Resonance Spectroscopy* ; Mitosis ; Necrosis ; Pathology* ; Prognosis ; Protons ; Tomography, Emission-Computed, Single-Photon*

PURPOSE: Thallim-201 (201Tl) brain SPECT, which can represent cellular activity of brain lesions, may provide more useful information in differentiating between benign and malignant brain lesions more so than CT or MRI, that merely represents anatomic changes or breakdown of blood brain barrier. We used 201Tl brain SPECT prospectively to evaluate the utility of 201Tl-indices as an indicator of benign or malignant lesions. MATERIALS AND METHODS: We studied 28 patients. There were 13 cases of benign lesions (3: nonspecific benign lesion, 3: meningioma, 2: low grade glioma, 1: tuberculoma, central neurocytoma, hemangioblastoma, radiation necrosis, and choroid plexus papilloma) and 15 cases of malignant lesions (6: glioblastoma multiforme, 5: anaplastic glioma, 2: medulloblastoma, 1: metastasis and lymphoma). In all patients, CT and/or MRI were obtained and then 201Tl brain SPECT was obtained with measuring mean 201Tl index and peak 201Tl index. An unpaired t-test was performed to compare the 201Tl-indices and pathologic diagnoses to evaluate the utility of 201Tl-indices as an indicator of benign or malignant lesions. RESULTS: There were no statistically significant difference in 201Tl-indices between benign and malignant brain lesions (P>0.05). CONCLUSION: These results demonstrated that we could not use 201Tl indices on brain SPECT alone as an indicator of benign or malignant brain lesions.
Blood-Brain Barrier ; Brain* ; Choroid Plexus ; Diagnosis ; Glioblastoma ; Glioma ; Hemangioblastoma ; Humans ; Magnetic Resonance Imaging ; Medulloblastoma ; Meningioma ; Necrosis ; Neoplasm Metastasis ; Neurocytoma ; Prospective Studies ; Tomography, Emission-Computed, Single-Photon* ; Tuberculoma

BACKGROUND: It has been reported that the association between blood pressure and cognitive impairment could be modified by factors such as gender, education, and Apo E genotypes. This study was aimed to investigate whether `age' could also be a modifier of the association. METHODS: The study group consisted of 489 participants aged 65 years or over who lived in Namwon, Korea. The presence of hypertension was ascertained by the measurement of their resting blood pressure. A cognitive impairment was considered to be present when a Korean Mini-Mental State Examination score was less than 19 (30th percentile). RESULTS: A higher systolic blood pressure was significantly associated with cognitive impairment, after an adjustment for age, gender, education, and occupation. However, the significance was lost in subjects aged 71 or over. CONCLUSIONS: High systolic blood pressure is a factor strongly associated with cognitive impairment in an aged population. However, in the subjects with far advanced age, the association is not significant. Further studies are required to elucidate whether blood pressure lowering strategies should be differentiated according to age in order to reduce cognitive impairment.
Adult* ; Aged* ; Apolipoproteins E ; Blood Pressure* ; Education ; Genotype ; Humans ; Hypertension ; Jeollabuk-do ; Korea ; Occupations

PURPOSE: The purpose of this study was to explore the direction for development of the Korean Journal of Adult Nursing toward becoming an international journal through analysing the accepted and rejected papers during the last three years (2007-2009). METHODS: Two hundred and ten accepted papers were analyzed focusing on research methodology and key words using descriptive statistics. In addition, rejected papers were reviewed to analyze their study designs and key words. RESULTS: The proportion of quantitative research was 86.4% while the proportion of qualitative research was 9.5%. The majority of the qualitative research design was survey (71.8%). Sixty percent of the research had verbal consent and 32.7% had written consent from the participants. The prevailing data collection settings were hospitals (52.1%), and community (22.7%). The most frequently used research domain was health. It was noted that theoretical framework was rarely presented. The paper rejection rate was 31.5% and among the rejected paper, 75.3% was survey. CONCLUSION: The results of this analysis suggest that published studies have been improved and diversified compared with the papers published before the year 2007. However, translation research, clinical trials by nurses, and more detailed evaluation process for ethics in research need to be facilitated.
Adult ; Data Collection ; Humans ; Nursing Research ; Qualitative Research ; Rejection (Psychology) ; Research Design

Background: Human breast milk is essential and provides irreplaceable nutrients for early humans. However, breastfeeding is not easy for various reasons in medical institution environments. Therefore, in order to improve the breastfeeding environment, we investigated the difficult reality of breastfeeding through questionnaire responses from medical institution workers. Methods: A survey was conducted among 179 medical institution workers with experience in childbirth within the last five years. The survey results of 175 people were analyzed, with incoherent answers excluded. Results: Of the 175 people surveyed, a total of 108 people (61.7%) worked during the day, and 33 people (18.9%) worked in three shifts. Among 133 mothers who stayed with their babies in the same nursing room, 111 (93.3%) kept breastfeeding for more than a month, but among those who stayed apart, only 10 (71.4%) continued breastfeeding for more than a month (P = 0.024). Ninety-five (88.0%) of daytime workers, 32 (94.1%) two-shift workers, and 33 (100%) three-shift workers continued breastfeeding for more than a month (P = 0.026). Workers in general hospitals tended to breastfeed for significantly longer than those that worked in tertiary hospitals (P = 0.003). A difference was also noted between occupation categories (P = 0.019), but a more significant difference was found in the comparison between nurses and doctors (P = 0.012). Longer breastfeeding periods were noted when mothers worked three shifts (P = 0.037). Depending on the period planned for breastfeeding prior to childbirth, the actual breastfeeding maintenance period after birth showed a significant difference (P = 0.002). Of 112 mothers who responded to the question regarding difficulties in breastfeeding after returning to work, 87 (77.7%) mentioned a lack of time caused by being busy at work, 82 (73.2%) mentioned the need for places and appropriate circumstances. Conclusion In medical institutions, it is recommended that environmental improvements in medical institutions, the implementation of supporting policies, and the provision of specialized education on breastfeeding are necessary to promote breastfeeding.