Proteomic analysis of differential protein expression in atherosclerosis.

Although recent studies have shown that several pro-inflammatory proteins can be used as biomarkers for atherosclerosis, the mechanism of atherogenesis is unclear and little information is available regarding proteins involved in development of the disease. Atherosclerotic tissue samples were collected from patients in order to identify the proteins involved in atherogenesis. The protein expression profile of atherosclerosis patients was analysed using two-dimensional electrophoresis-based proteomics. Thirty-nine proteins were detected that were differentially expressed in the atherosclerotic aorta compared with the normal aorta. Twenty-seven of these proteins were identified in the MS-FIT database. They are involved in a number of biological processes, including calcium-mediated processes, migration of vascular smooth muscle cells, matrix metalloproteinase activation and regulation of pro-inflammatory cytokines. Confirmation of differential protein expression was performed by Western blot analysis. Potential applications of the results include the identification and characterization of signalling pathways involved in atherogenesis, and further exploration of the role of selected identified proteins in atherosclerosis.

Trichomonas vaginalis-induced apoptosis in RAW264.7 cells is regulated through Bcl-xL, but not Bcl-2.

The purpose of this study was to determine whether anti-apoptotic proteins of the Bcl-2 family such as Bcl-2 and Bcl-x(L), proteins that confer resistance to apoptotic death from some stimuli, block apoptotic cell death in RAW264.7 cells upon treatment with Trichomonas vaginalis. In this study, the expression level of Bcl-2 was unchanged throughout the course of apoptotic cell death, and overexpressed Bcl-2 did not prevent release of cytochrome c, the significant change of the membrane potential, activation of caspases, and PARP cleavage in T. vaginalis-treated RAW264.7 cells. On the other hand, Bcl-x(L)expression was decreased after T. vaginalis treatment accompanied with Bax activation. Furthermore, we showed that release of mitochondrial cytochrome c, cleavage of caspase-9 and PARP during apoptosis in T. vaginalis-treated RAW264.7 cells were considerably diminished by transfection with overexpressed Bcl-x(L), and overexpressed Bcl-x(L)could inhibit T. vaginalis-induced apoptosis in RAW264.7 cells. In addition, interestingly, pre-treatment with caspase inhibitors, Boc-D-FMK and Z-DEVD-FMK, significantly abolished T. vaginalis-induced down-regulation of Bcl-x(L), suggesting that caspase-3 may play a pivotal role in the process of apoptosis as well as the down-regulation of Bcl-x(L)by T. vaginalis. Therefore, these results suggest that T. vaginalis-induced apoptosis in RAW264.7 cells can occur via a Bcl-x(L)-dependent apoptotic mechanism.

Susceptibility of avian hosts to experimental Gymnophalloides seoi infection.

To determine whether avian species are susceptible to infection with Gymnophalloides seoi (a human-infecting intestinal trematode), we exposed 7 species of birds with metacercariae obtained from oysters. The birds were necropsied at days 2, 4, and 6 postinfection (PI). The highest worm recovery at day 6 PI was obtained from the Kentish plover (Charadrius alexandrinus; mean = 56.0%), followed by the Mongolian plover (C. mongolus; 49.3%), and the grey plover (Pluvialis squatarola; 32.3%). In contrast, no mature worms were recovered from the great knot (Calidris tenuirostris), dunlin (C. alpina), black-tailed gull (Larus crassirostris), and mallard (Anas platyrhynchos). Among the plovers, the worms attained the greatest size at day 6 PI (254.1 x 190.4 microm) in the Kentish plover, with a significantly higher number of eggs in the uterus. The 3 species of plovers are highly susceptible to experimental G. seoi infection, suggesting that they could play a role as definitive hosts for these worms in nature.

Enterobius vermicularis egg positive rates in primary school children in Gangwon-do (province), Korea.

A survey of the infection rate of Enterobius vermicularis among students in 4 primary schools located in Gangwon-do (Province) was done from May to June 2001. Among the 398 examinees, 39 (9.8%) were infected with E. vermicularis demonstrated by the adhesive cellotape anal swab method. The infection rates ranged from 8.3% to 11.8% among the four schools. The infection rate of males and females was 10.7% and 7.7% respectively. The first grade students showed the highest infection rate, 28.7%. The confirmed cases were treated with albendazole three times at an interval of 15 days. We were able to confirm that E. vermicularis infection is still prevalent among students in Gangwon-do, Korea.

Mucosal mast cell responses in the small intestine of rats infected with Echinostoma hortense.

Mucosal mast cell (MMC) responses and worm recovery rates in rats infected with Echinostoma hortense were investigated from day 3 to day 56 post-infection (p.i.). Experimental infected group showed apparently higher number of MMC in each part of the small intestine than that of the control group. The number of MMC in the duodenum increased gradually after the infection and reached a peak on day 35 p.i. Thereafter, the number of MMC continued to decrease at a slow pace. The kinetics of MMC responses in the upper and lower jejunum were similar to that of the duodenum, but the number of MMC in the jejunum was lower. The worm recovery rate decreased with respect to time of which it was markedly reduced on day 49 and 56 p.i. The duration in which a high number of MMC appeared was similar to that in which a low rate in worm recovery was recorded. These results indicate that intestinal mastocytosis may play an important role in the expulsion of E. hortense.

The palearctic oystercatcher Haematopus ostralegus, a natural definitive host for Gymnophalloides seoi.

In order to verify the role of migrating birds as natural definitive hosts for Gymnophalloides seoi (Gymnophallidae), Palearctic oystercatchers, Haematopus ostralegus, were caught from several western coastal areas in the Republic of Korea and examined for intestinal flukes. Five (71.4%) of 7 oystercatchers were infected with 302-1,660 (mean 892) adult G. seoi. In intestinal sections of the host, worms were found in the intervillous space of the mucosal layer. We conclude that the oystercatcher is a natural definitive host for G. seoi.

An incidental case of human Heterophyes nocens infection diagnosed by sectional morphology in a biopsy specimen of the small intestine.

A case of human infection with Heterophyes nocens (Heterophyidae) was incidentally found in a biopsy specimen of the Meckel's diverticulum at the upper part of the small intestine. The patient was a 58-year-old man living in a rural area of Talsonggun, Kyongsangbuk-do. He had gastrointestinal symptoms such as epigastric pain, indigestion, and abdominal discomfort for 3 months, and severe diarrhea, abdominal pain, and vomiting for about 1 month before hospitalization. Endoscopy of the upper part of the small intestine revealed a Meckel's diverticulum, and it was excised and histopathologically examined. Three adult flukes were incidentally found sectioned in the mucosa, and they were identified as H. nocens. The patient had a history of eating raw mullets at a fish market in Pusan 6 months ago, and the mullets were presumed to be the source of infection. This case brings a considerable interest in that specific diagnosis of heterophyid infections could be done by sectional morphology of the worms.

Discovery of Gymnophalloides seoi metacercariae in oysters from islands of the West Sea known as the habitats of paleartic oystercatchers.

An epidemiological survey was performed to know the infection status of oysters with Gymnophalloides seoi metacercariae in 7 islands of the West Sea known as the habitat of paleartic oystercatchers, Haematopus ostralegus osculans, in Korea. The surveyed areas were Aphaedo (Shinan-gun, Chollanam-do), Jangjado, Sonyudo and Munyodo (Okdo-myon, Kunsan-shi, Chollabuk-do), Yubudo (Changhang-up, Sochon-gun Chungchongnam-do), and Polumdo and Chumoondo (Sodo-myon, Kangwha-gun. Inchonshi). The oysters collected from Aphaedo, the known endemic focus, were examined monthly from August 1995 to October 1996 for observation of any seasonal variation of the metacercarial density. The average metacercarial burden was 761-2,077 by month, but the seasonal variation of the metacercarial density was not obvious. A total of 54 metacercariae was detected in 63 oysters collected from Yubudo. Out of 30 oysters from Sonyudo, 25 (83.3%) were infected with 1-66 metacercariae (12.6 in average). All of 50 oysters (100%) from Munyodo were infected with 3-162 metacercariae (53.5 in average). Only 4 metacercariae were detected in 100 oysters from Chumoondo. However, no metacercariae were found in 55 oysters from Jangjado and 50 oysters from Polumdo. From the above results, it was confirmed that G. seoi is still highly prevalent in oysters from Aphaedo, and several islands of the West Sea known as the habitats of paleartic oystercatchers are new endemic areas of this fluke.

Two rare cases of Diphyllobothrium latum parvum type infection in Korea.

Two rare cases of human infection with parvum (dwarf) type of Diphyllobothrium latum [syn. D. parvum (Stephens, 1908)], were discovered in Korea. The first case was a 46-year old housewife, from whom a kind of pseudophyllidean tapeworm eggs was detected in the feces. She was treated with praziquantel and purged, and a complete strobila with scolex, 120 cm in total length, was recovered. She recalled that she had eaten raw trouts at a raw-fish restaurant near the Chungju Lake. Another patient was a 22-year old medical student (male), who used to eat raw sea-foods. He discovered a chain of tapeworm proglottids, 15 cm in length, discharged spontaneously in his stool and brought it for identification. The worms from the two cases were compatible with D. parvum (Stephens, 1908) of which the taxonomic significance has long been questioned. After a detailed morphological study and review of literature, we designated the worms as D. latum parvum type. This is the first report on the occurrence of this rare type of D. latum infection in Korea.

[Trematode infections in the small intestine of Egretta alba modesta in Kangwon-do].

We surveyed 12 migrating large egrets, Egretta alba modesta, for their infection status with intestinal trematodes, from June to September, 1990. All of them were infected with one or more flukes. Total 5 species were identified. Their infection rates and total numbers of worms were Metagonimus sp. 67% and 1,412, Centrocestus armatus 67% and 2,306, Echinochasmus japonicus 50% and 99, Strigea falconis 83% and 635, and a diplostomid fluke 33% and 36. The number of worms in a bird was in the range from 6 to 896. This is the first record that the egret is a reservoir host of Metagonimus sp., C. armatus, E. japonicus, and S. falconis. S. falconis is a new parasite fauna in Korea. Migrating birds must play an important role in the transmission and spreading of these intestinal trematodes in Korea.

[Studies on Echinostoma spp. in the Chungju Reservoir and upper streams of the Namhan River].

The present study was performed to know the epidemiological status of echinostomiasis in the Chungju Reservoir and upper streams of Namhan River, together with an experimental study on the life history of Echinostoma hortense. The stool specimens of 169 inhabitants and 473 junior high school students from 5 different villages revealed 3 (0.5%) echinostomatid egg positive cases. E. hortense adult worms were recovered from one patient after a treatment and purgation. For the other two patients, it was presumed that one had E. hortense and another E. cinetorchis infection, based on the morphology of eggs. Five kinds of freshwater snails (168 Radix auricularia coreana, 534 Physa acuta, 144 Hippeutis cantori, 56 Cipangopaludina chinensis malleata and 125 Semisulcospira nodifila globus) examined for the cercariae of echinostomes showed negative results. Ten kinds of freshwater fishes examined for E. hortense metacercariae revealed positive rates as Misgurnus anguillicaudatus 40.5%, Odontobutis obscura interrupta 20.3%, Moroco oxycephalus 3.9%, and Coreoperca kawamebari 2.0%. In the experimental study, the metacercariae of E. hortense were infected to rats, eggs were collected from adult worms and cultivated, and miracidia were obtained. The miracidia were artificially infected to freshwater snails (R. auricularia), and cercarial shedding was studied. It was revealed that, when the snails were kept at a low temperature (24 degrees C), only 523 cercariae (on average) were produced during 24 hrs, while they were at a high temperature (30 degrees C), as many as 9,990 cercariae (on average) were shed during the same time. The experimental infection of E. hortense cercariae to freshwater fishes was successful in O. obscura interrupta 52.0%, M. anguillicaudatus 30.3%, C. kawamebari 27.0%, Cobitis lutheri 15.0%, M. oxycephalus 7.3%, Pseudogobio esocinus 4.3%, Squalidus coreanus 2.0%, Zacco platypus 1.3%, and Pungtungia herzi 1.3%. However, infection was not successful to snails, C. chinensis. It has been proved that the Chungju Reservoir and upper streams of Namhan river are endemic areas of echinostomiasis, especially of E. hortense, and snails such as R. auricularia coreana and fishes such as O. obscura interrupta, M. anguillicaudatus, and others are taking the role of first and second intermediate hosts, respectively.

[Cercarial shedding of Echinostoma cinetorchis and experimental infection of the cercariae to several kinds of snails].

The development of Echinostoma cinetorchis in several snail species reared in laboratory aquaria was observed. The eggs from adult flukes collected from the intestine of rats were cultivated to miracidia, and exposed to Hippeutis sp. snails. Observations were made for cercarial shedding from the exposed snails. The cercariae shed from the snails were again exposed to several species of fresh water snails in order to observe metacercarial formation in the snails and their infectivity to final hosts. The results obtained in this study were as follows: 1. Twenty miracidia were exposed to each snail of Hippeutis sp. About 58.3% of the above snails (7 out of 12) were dead before shedding the cercariae, and the remainder shed the cercariae for a period of 7 to 9 days before death. 2. Cercarial shedding from the infected snails started from the 25th day after the exposure to miracidia, and the total number of cercariae shed per snail was 684 in average (range; 482-904). 3. The size of rediae developed in the infected Hippeutis sp. snails was 1,242 x 214 microns in average, and the number of rediae per snail was 350 in average (range; 120-510). 4. About 40 to 50 cercariae shed from the Hippeutis sp. snails were each exposed to several species of snails reared in the laboratory. The metacercarial formation was confirmed by dissecting the infected snails, 12 to 16 days after the infection.(ABSTRACT TRUNCATED AT 250 WORDS)