[Evaluation of the fabrication deviation of a kind of milling digital implant surgical guides].

OBJECTIVE: To evaluate the deviation of digital implant surgical guides during fabrication process in the Organical Dental Implant (ODI) system. METHODS: This study included two parts. The first part was the in vitro study. A resin block with a diagnostic template was used for the planning. After cone beam computed tomography (CBCT) scanning, a surgical guide with eight implants was virtually designed using the ODI system. The guide was milled by a 5-axial numerical controlled milling machine, and an optical scanning was taken to digitalize the guide to a standard tessellation language (STL) form. The STL data were then imported into an ODI software and registered with the original design. The deviation of the sleeves between the design and the STL was measured in the ODI software and set as the golden standard. Then the ODI examination table was used to measure the deviation of the guide during fabrication. Examiners A and B measured 10 times separately. The reliability and the validity of the examination table was calculated. The second part was the in vivo study: The deviation during fabrication of 12 guides designed and fabricated by the ODI system were measured using the examination table. RESULTS: The standard deviation of the deviation measured using the examination table by examiners A and B were all below 0.40 mm (for the shell reference points) and 0.71 degree (for the angles). No significant difference was found between the two examiners for any implant sites. The result of the examination table was larger than that of the software for the shell reference point (t-test, P<0.05), but no significant difference was found for the angle deviation (t-test, P>0.05). The 45 implants positions in the 12 guides for the in vivo study were examined using the examination table. The deviations at the shell reference points were (1.06±0.29) mm (0.42-1.75 mm), and at the implant tip were (1.12±0.48) mm (0.41-2.44 mm). The angle deviations were (1.42±0.70) degree (0.29-2.96 degree). CONCLUSION: Deviation is unavoidable during the fabrication process of the guides. The examination table of the ODI system is a reliable and valid tool to measure the deviation during fabrication of the ODI guides. More studies should be designed to research the relationship between the fabrication deviation and the implant insertion deviation.

Prevalence of tet gene and complete genome sequencing of tet gene-encoded plasmid (pAHH01) isolated from Aeromonas species in South Korea.

AIMS: To investigate the tetracycline resistance related to tet genes in Aeromonas isolates collected from water and diseased fish in South Korea. METHODS AND RESULTS: A total of 34 Aeromonas strains were examined for their susceptibility to tetracycline using the minimum inhibitory concentration (MIC) assay, and the genetic determinants (tetA to E) were analysed. Among these strains, the tetA and tetE genes were predominant (tetA was found in six strains, and tetE was found in nine strains), and 15 strains were tetracycline-resistant by the MIC assay. Additionally, the 8979-bp plasmid that contains the tetE gene was fully sequenced. CONCLUSIONS: These data may be important with regard to the spread and persistence of tetracycline resistance genes in the bacterial populations that are present in aquaculture systems. SIGNIFICANCE AND IMPACT OF THE STUDY: Interestingly, no isolate has previously been shown to harbour three tet genes that are mediated by efflux systems, but the tetA, tetD and tetE genes were all isolated from one strain, which had the highest MIC value for tetracycline among the strains analysed in this study. We also investigated the full-length plasmid that encoded the tetE gene from a tetracycline-resistant strain.

First description of ColE-type plasmid in Aeromonas spp. carrying quinolone resistance (qnrS2) gene.

AIMS: Isolation and full sequence analysis of ColE-type plasmid, which carries the qnrS2 gene. METHODS AND RESULTS: Quinolone resistance (qnrS2) gene-carrying plasmids were isolated from Aeromonas sobria and Aeromonas hydrophila strains, and plasmid sequencing was achieved by a primer-walking approach. The total sizes of these plasmids (pAQ2-1 and pAQ2-2) were 6900 bp and 6903 bp, respectively, and they were 99·1% identical to each other. The genes (oriV and repA) for plasmid replication were organized similar to the corresponding genes in the ColE2-type plasmids, pAsa3 and pAsa1, isolated from Aeromonas salmonicida subsp. salmonicida, but the gene (mobA) for mobilization was homologue to ColE1-type plasmid (pAsa2) from Aer. salmonicida subsp. salmonicida. Additionally, the qnrS2 gene was part of a mobile insertion cassette element in the plasmid. CONCLUSIONS: Two plasmids were assumed to be the same plasmid, and this identification of a plasmid-mediated qnrS2 gene from the two different strains underlines a possible diffusion of these resistance determinants in an aquaculture system. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first finding of the ColE-type plasmid carrying the qnrS2 gene.

A small IncQ-type plasmid carrying the quinolone resistance (qnrS2) gene from Aeromonas hydrophila.

AIMS: To investigate the qnrS2 gene encoded by a plasmid obtained from Aeromonas hydrophila. METHODS AND RESULTS: To investigate the full-length sequence of the plasmid carrying qnrS2 (plasmid designated pAHH04) from the strain SNUFPC-A10, the full-length coding sequence of the qnrS region was first amplified. The remaining part of the plasmid was read outwards from this region. The plasmid pAHH04 contained the repC, repA, mobA and mobC genes, and its total size was 7191 bp with a G+C content of 60%. CONCLUSIONS: This study describes the full-length sequence of a plasmid carrying the qnrS2 gene from Aer. hydrophila. The plasmid pAHH04 carried plasmid replication and mobilization genes from IncQ-type plasmids. SIGNIFICANCE AND IMPACT OF THE STUDY: The isolated qnrS2 gene encoded by a plasmid from an Aer. hydrophila strain is of significant importance because it emphasizes the problem of antibiotic resistance as well as the ability of the determinants to spread among the different bacterial species that impact human health.

Hypoxia appears at pre-arthritic stage and shows co-localization with early synovial inflammation in collagen induced arthritis.

OBJECTIVE: The presence of hypoxia in rheumatoid synovium has been well known, but exact correlation between hypoxia and synovitis is unclear. The aim of our study was to investigate the time and spatial relationship and the correlation of severity between hypoxia and synovitis in pre-arthritic or early stage of inflammatory joint disease. METHODS: DBA/1J mice were injected intradermally with type II collagen and adjuvant solution to induce arthritis; mice injected with only adjuvant were used as a control group. CIA and control mice were sacrificed weekly after the injection to evaluate serial pathological changes. H&E stain and hydroxyprobe-1 stain were performed to look at the status of inflammation and hypoxia. RESULTS: In serial observations of tissue pathology, we could note the inflammation of synovium developing a week after the injection of type II collagen. Hypoxic change, measured by the hydroxyprobe-1 stain, was also identified in synovium as early as 1 week after the collagen injection, prior to clinically evident arthritis. In addition, we could observe that inflammation and hypoxia co-localize in the synovium and there was a positive correlation between the severity of hypoxia and the degree of synovitis. CONCLUSION: Our results demonstrate that hypoxia takes place in synovium at the pre-arthritic stage of disease and have a close spatial relationship and a positive severity correlation with synovitis.

A human case of Plagiorchis vespertilionis (Digenea: Plagiorchiidae) infection in the Republic of Korea.

Plagiorchis vespertilionis (Digenea: Plagiorchiidae) is generally considered a bat parasite, but here it is reported for the first time in a human. The patient was a 34-yr-old male who lived in a coastal village of Haenam-gun (county), Jeollanam-do (province), Republic of Korea. Only 1 worm, 2.6 mm long and 0.7 mm wide, was recovered after praziquantel treatment and purging with magnesium salts. The fluke was characterized by a large body size, a sucker ratio of 1:1, a straight cirrus organ, a short distance between the ventral sucker and ovary, well-developed vitellaria, a uterus with descending and ascending loops, and fully developed eggs with an average size of 32.5 x 17.5 microm. The patient had habitually eaten the raw flesh of snakehead mullet and gobies that had been caught near his village. The present case represents the first record of a human P. vespertilionis infection.

Detection of LGI1 and CASPR2 antibodies with a commercial cell-based assay in patients with very high VGKC-complex antibody levels.

BACKGROUND: The presence of VGKC-complex antibodies, without LGI1/CASPR2 antibodies, as a standalone marker for neurological autoimmunity remains controversial. Additionally, the lack of an unequivocal VGKC-complex antibody cut-off level defining neurological autoimmunity makes it important to test for monospecific antibodies. We aim to determine the performance characteristics of a commercial assay (Euroimmun, Lübeck, Germany) for LGI1/CASPR2 antibody detection in patients with very high VGKC-complex antibody levels and report their clinico-serological associations. METHODS: We identified 8 patients in our cohort with the highest VGKC-complex antibody levels (median 2663.5pM, range 933-6730pM) with VGKC-complex antibody related syndromes (Group A). Two other groups were identified; 1 group with suspected neuronal surface antibody syndromes and negative for VGKC-complex antibodies (Group B, n=8), and another group with cerebellar ataxia and negative for onconeuronal antibodies (Group C, n=8). RESULTS: Seven out of 8 patients (87.5%) in Group A had LGI1 and/or CASPR2 antibodies. One Group B patient had LGI1 antibodies but was negative on re-testing with a live cell assay. No Group C patients had monospecific antibodies. Inter-rater reliability was high; combining Groups A and B patients, the kappa statistic was 0.87 and 1.0 for LGI1 and CASPR2 antibodies respectively. CONCLUSION: We demonstrated that a high proportion of patients with very high VGKC-complex antibody levels and relevant clinical syndromes have LGI1 and/or CASPR2 antibodies detected by the commercial assay. Our findings lend support to the use of the assay for rapid and reliable detection of LGI1 and CASPR2 antibodies.

A Case of Ectopic Paragonimiasis in a 17th Century Korean Mummy.

Archaeoparasitological studies on fossilized feces obtained from Joseon Dynasty (1392-1910 CE) mummies have provided invaluable data on the patterns of parasitic infection in pre-modern Korean societies. In our recent radiological investigation of a 17th century Joseon mummy discovered in Cheongdo (South Korea), we located a liver mass just below the diaphragm. Anatomical dissection confirmed the presence of a mass of unknown etiology. A subsequent parasitological examination of a sample of the mass revealed a large number of ancient Paragonimus sp. eggs, making the current report the first archaeoparasitological case of liver abscess caused by ectopic paragonimiasis.

Radioresistance of Anisakis simplex third-stage larvae and the possible role of superoxide dismutase.

The radioresistance of Anisakis simplex third-stage larvae and the possible role of sublethal radiation on superoxide dismutase (SOD) were investigated. Larvae were isolated from the viscera of the sea eel Anago anago; irradiated with 10, 100, 200, 500, or 1,000 Gy; and then given orally to rats. Worms were recovered at 16 hr postinoculation. Most larvae were found to have invaded the gastric wall, omentum, and abdominal cavity, suggesting that their viability and infectivity were not controlled by irradiation with the doses used. To determine the relationship between SOD activities in parasites and their radiosensitivities, the larvae of A. simplex and the metacercariae of Neodiplostomum seoulense (a radiosensitive control) were irradiated with 0, 30, 100, or 500 Gy, and parasite SOD levels were measured. In nonirradiated A. simplex larvae, the average SOD level was 38.9 U/mg, and this increased to 51.3 U/mg at 500 Gy. However, at all radiation doses applied, SOD activities of N. seoulense metacercariae were significantly (P < 0.05) lower than those of A. simplex larvae. Our results demonstrate that A. simplex third-stage larvae are radioresistant, and suggest that SOD plays a role in this radioresistance.

Suppressed cytokine and immunoglobulin secretions by murine splenic lymphocytes infected in vitro with Toxoplasma gondii tachyzoites.

Mechanisms of host immunosuppression after infection with Toxoplasma gondii are unclear. This study was performed to observe cytokine and immunoglobulin secretions by murine splenic lymphocytes infected in vitro with live, nonreplicating (irradiated) RH tachyzoites on stimulation with concanavalin A (Con A) or lipopolysaccharide (LPS). For lymphocyte cultivation, 3 groups were prepared: coculture with live nonirradiated tachyzoites separated by a transwell (group T), live irradiated tachyzoites without a transwell (group R), and no tachyzoites (group C). Compared with group T, groups R and C, on stimulation with Con A, revealed significantly (P < 0.05) lower levels of interleukin-2 (IL-2) and IFN-gamma, but not IL-10. The levels of IgG1, IgG2a, IgG2b, IgG3, IgA, and IgM were also significantly (P < 0.05) lower in groups R and C than in group T after stimulation with LPS. The results suggest that intracellular infection of murine splenic lymphocytes with T. gondii tachyzoites could impair their capacity to produce cytokine and immunoglobulin secretions.

Comparative natural killer cell activities of thymic, bursal, splenic and intestinal intraepithelial lymphocytes of chickens.

The natural killer (NK) cell activities of spleen, thymus, bursa, peripheral blood and gut intraepithelial lymphocytes (IEL) from FP and SC chickens were investigated in 4-hr and 16-hr 51Cr release assays. Target cells were 4 different tumor cell lines derived from either an avian leukosis tumor transplant (LSCC-RP9, LSCC-RP12) or from Marek's disease lymphomas (MDCC-MSB-1, MCDD-CU36). Great variability in cytotoxic potential was observed among NK cells of different lymphoid organs. NK cell cytotoxicity varied depending upon the type of effector cells, type of target cells, the ratio of effector to target cells, and the age and genetic background of chickens. Substantial levels of NK cell activity were detected in spleen and gut IEL of SC chickens in a 4-hr assay. In contrast, the NK cytotoxicity in gut IEL of FP chickens was not detectable until 16 hr after incubation. The ranges of target cell specificity demonstrated by IEL, spleen, thymus and bursa NK cells were similar to one another and, in general, the level of cytotoxicity increased with incubation time. Thymus and bursa NK cell activity of both SC and FP chickens was not detectable in a 4-hr assay but substantial NK cell activity was demonstrated in a 16-hr assay. The results of the present study demonstrate that various lymphoid organs of chickens, such as spleen, thymus, bursa, and gut intraepithelium, contain subpopulations of cells that can mediate spontaneous cytotoxicity.

Enhancement of dimethylnitrosamine-induced glutathione S-transferase P-positive hepatic foci by Clonorchis sinensis infestation in F344 rats.

The modifying potential of Clonorchis sinensis (CS) infestation on dimethylnitrosamine (DMN)-induced hepatocarcinogenesis was studied in male F344 rats. The metacercariae of CS were infected, 60 to each animal, by a single intragastric intubation at various times, before, during or after an 8-week period of DMN (25 ppm in drinking water) treatment. Controls were treated with DMN alone or infected with CS without carcinogen. Animals killed at the end of the 40-week experimental period showed a significant enhancing effect of CS on GST-P+ foci induction when CS was infected 4 weeks before DMN treatment, although no such influence was evident with CS infection during or following DMN exposure. The present findings suggest that CS might facilitate the proliferation of DMN-induced preneoplastic lesions of liver in rats.

A trial for a DNA diagnosis of Plasmodium vivax malaria recently reemerging in the Republic of Korea using microtiter plate hybridization assay.

The polymerase chain reaction-based microtiter plate hybridization (PCR-MPH) assay was utilized for a DNA diagnosis of Plasmodium vivax malaria, which has recently reemerged in the Republic of Korea. The subjects were 18 parasite-proven patients and 5 healthy controls. Follow-up blood samples were collected from 4 patients after a standard course of treatment. Polymerase chain reaction and electrophoresis of all the patients' blood showed a prominent band at the 138 base pair area, but not in the controls or after treating the patients. Hybridization of the PCR products with known species-specific probes of the 18S rRNA of various malaria species revealed strong positive reactions against the Plasmodium vivax-specific probe (absorbance 1.30-1.90 at 405 nm) in all of the patients. The absorbance was positively correlated with the degree of blood parasitemia, but with a borderline significance. Sequencing of the probe region of the Korean P. vivax revealed no significant variations from the typical P. vivax. The results show that the PCR-MPH is a highly useful technique for the DNA diagnosis of Korean vivax malaria.

Hepatic capillariasis: first case report in the Republic of Korea.

We report a case of massive hepatic infection by Capillaria hepatica in a 14-month-old girl who presented with the symptom triad of persistent fever, hepatomegaly, and leukocytosis with eosinophilia. Twenty-five cases of human infection with this parasite, mostly in children, have been reported in the literature. This is the first case of hepatic capillariasis reported in the Republic of Korea. The diagnosis was made by needle biopsy of the liver. Scanning electron microscopic examination of the biopsy specimen was also performed. Thiabendazole therapy was initiated and the patient developed liver disease-related IgA nephropathy during the therapy. The literature dealing with proven cases of infection with C. hepatica is briefly reviewed.

Clinical features of vivax malaria.

Plasmodium vivax malaria reemerged in the Republic of Korea in 1993 near the Demilitarized Zone (DMZ). We reviewed clinical features of 101 symptomatic patients with vivax malaria. Of the patients, 77 patients (76.3%) were veterans who had served near the DMZ; their median age was 23 years. The duration of the minimum latent period was > 6 months in 66.2% (51 of 77) of the patients (median, 278 days). Tertian fever developed in 69 patients (68.3%). Severe thrombocytopenia with platelet counts < 60,000/microL was common (29.6% of patients). The parasite densities ranged 32-52,127 parasites per microliter of blood (geometric mean, 1,287). The only complication was a splenic rupture in one patient. All patients responded promptly to chloroquine therapy. Our data suggest that the clinical features of reemerging vivax malaria may be similar to those of Korean vivax malaria reported in the past.

High prevalence and seasonality of cryptosporidiosis in a small rural village occupied predominantly by aged people in the Republic of Korea.

The prevalence of Cryptosporidium parvum infection and its seasonality were studied in a small rural village occupied predominantly by aged people in Hwasun-gun, Chollanam-do, Republic of Korea. Fecal samples were collected monthly from November 1996 to October 1997 and examined for enteric parasites. To detect oocysts of C. parvum, the modified acid-fast stain was applied. To determine effects of climatic factors, local weather reports were collected. The overall oocyst prevalence during 1 year was 57.0% (77 of 135), and was significantly higher (P < 0.0001) in aged people--those aged 50-59 years (80.0%) and those aged 60-69 years (69.0%)--than in younger groups. No significant difference (P > 0.05) was noted between men and boys (53.6%) and women and girls (59.5%), but significant (P < 0.05) household clustering was recognized. People aged > 50 years tended to be more repeatedly positive than people in the younger group. The rate varied monthly from 7.8 to 20.0% (average, 12.6%), with significantly higher (P < 0.05) rates in spring than in summer and winter months. This seasonality had little relation with temperature (r = 0.04), but a weak correlation was found with rainfall (r = 0.4). It is of special interest that cryptosporidiosis is highly endemic among the aged population.

A case of intestinal capillariasis in the Republic of Korea.

In 1991, we observed a Korean man infected by Capillaria philippinensis who had no history of traveling in known endemic areas. He had year-long diarrhea with severe loss of body weight. He enjoyed hunting and fishing and eating raw meat of game animals and fish. An open full-thickness biopsy of the ileum showed flat mucosal surface and sections of the round worm, and fecal examination revealed numerous elliptical helminth eggs. The worms and eggs were consistent with features of C. philippinensis. He was treated with albendazole and was cured. This is the first known case of intestinal capillariasis in the Republic of Korea.

High prevalence of Gymnophalloides seoi infection in a village on a southwestern island of the Republic of Korea.

Gymnophalloides seoi (Digenea: Gymnophallidae), a new human intestinal trematode reported from a Korean woman who complained of epigastric discomfort, has been shown to be highly prevalent among the villagers of a southwestern island of the Republic of Korea. For the detection of human infections, fecal examinations were conducted on the inhabitants of a seashore village, where the first patient with a G. seoi infection had resided. Of 98 inhabitants examined, 70 (71.4%) were infected with various intestinal parasites; among them, G. seoi showed the highest rate of egg positivity 48 of 98 (49.0%). Individual worm burdens of G. seoi, as measured by collection of adult flukes after anthelmintic treatment and purgation, ranged from 106 to 26,373 specimens (average per infected case = 3,326). There was no sex-related difference in the prevalence of G. seoi, and the age distribution of the infected cases showed a relatively even pattern. This study confirms that human infection by G. seoi is not an incidental one and provides the first evidence of its high prevalence on this Korean island.

Automatic identification of human helminth eggs on microscopic fecal specimens using digital image processing and an artificial neural network.

In order to automate routine fecal examination for parasitic diseases, we propose in this study a computer processing algorithm using digital image processing techniques and an artificial neural network (ANN) classifier. The morphometric characteristics of eggs of human parasites in fecal specimens were extracted from microscopic images through digital image processing. An ANN then identified the parasite species based on those characteristics. We selected four morphometric features based on three morphological characteristics representing shape, shell smoothness, and size. A total of 82 microscopic images containing seven common human helminth eggs were used. The first stage (ANN-1) of the proposed ANN classification system isolated eggs from confusing artifacts. The second stage (ANN-2) classified eggs by species. The performance of ANN was evaluated by the tenfold cross-validation method to obviate the dependency on the selection of training samples. Cross-validation results showed 86.1% average correct classification ratio for ANN-1 and 90.3% for ANN-2 with small variances of 46.0 and 39.0, respectively. The algorithm developed will be an essential part of a completely automated fecal examination system.

Histopathological changes of the cat brain in experimental sparganosis.

Histopathological changes of the brain were observed in 22 mongrel cats each of which was subjected to intracranial inoculation of 3 or 5 plerocercoids of Spirometra erinacei. The brains were examined at 2 weeks, 1, 3, and 6 months post-inoculation (PI). Grossly, the brain lesion consisted of a mass of worm capsule and/or hemorrhagic tract. Microscopically hypertrophied astrocytes, lymphocytes, plasma cells, RBCs, polymorphonuclear leukocytes, and hemosiderin laden macrophages were found around the worms or tracts in all groups. The number of reactive astrocytes increased mainly in the white matter. In the lesions 3 months PI, the sectioned worms were encircled with fibrous tissue along with numerous RBCs, inflammatory cells, and necrotic debris. Collagenous fibers were also demonstrated. Outside the inflammation, a wide edematous zone developed and numerous neuroglial fibers were also found. The location of the worms or the tracts was random in the brain lobes, but the worm seemed to migrate mainly along the white matter.