BACKGROUND: Gastric carcinomas (GCs) have recently been reclassified according to the mucin phenotypes. We aimed to characterize the relationship between the mucin phenotypes and the genetic alterations or the clinicopathologic parameters of GCs. METHODS: Immunohistochemistry was performed for MUC1, MUC5AC, MUC6, MUC2, CD10, p53, hMLH1, CerbB2 and E-cadherin in 150 GCs. The mucin phenotypes of the GCs were classified as 4 phenotypes: gastric, intestinal, mixed and unclassified. RESULTS: MUC1, MUC5AC, MUC6, MUC2 and CD10 were expressed in 63.3%, 42.7%, 14.0%, 24.7% and 14.0% of the GCs, respectively. The mucin phenotypes of the GCs corresponded to the gastric type in 31.3%, the intestinal type in 20.0%, the mixed type in 15.3% and the unclassified type in 33.3%. The incidence of a p53 overexpression was higher in the gastric or mixed phenotype than in the intestinal or unclassified phenotype. MUC5AC expression, p53 overexpression and the gastric or mixed phenotype were associated with poor patient survival by multivariate analysis. CONCLUSION: This study suggests the gastric or mixed mucin phenotype may more likely go through the p53 pathway in carcinogenesis and the mucin phenotype may be considered as a prognostic indicator.
Cadherins ; Carcinogenesis ; Humans ; Immunohistochemistry ; Incidence ; Mucins* ; Multivariate Analysis ; Phenotype* ; Stomach ; Tumor Suppressor Protein p53

In an effort to characterize, on the molecular scale, the Acanthamoeba initially isolated from the cornea of an amoebic keratitis patient associated with overnight-wear orthokeratology lens in Korea, we conducted mitochondrial DNA restriction fragment length polymorphism, 18S rDNA sequencing, and drug sensitivity analyses on the isolate (KA/PE1). The patient was treated with polyhexamethylene biguanide, chlorhexidine and oral itraconazole, which resulted in resolution of the patient's ocular inflammation. The majority of the molecular characteristics of the KA/PE1 were determined to be identical, or quite similar, to those of A. castellanii Ma strain, which had been isolated also from amoebic keratitis. The risk of Acanthamoeba keratitis as a potential complication of overnight orthokeratology is briefly discussed.
*Sequence Analysis, DNA ; RNA, Ribosomal, 18S/genetics ; Polymorphism, Restriction Fragment Length ; Parasitic Sensitivity Tests ; Myopia/therapy ; Itraconazole/administration & dosage ; Humans ; Female ; Disinfectants/administration & dosage ; DNA, Ribosomal/analysis ; DNA, Protozoan/analysis ; DNA, Mitochondrial/analysis ; Contact Lenses/*adverse effects ; Chlorhexidine/administration & dosage ; Biguanides/administration & dosage ; Astigmatism/therapy ; Antiprotozoal Agents/administration & dosage ; Animals ; Adolescent ; Acanthamoeba Keratitis/drug therapy/*parasitology ; Acanthamoeba/classification/*genetics/*isolation & purification

INTRODUCTIONTo investigate the attitudes of medical and nursing student towards the older Chinese population.

MATERIALS AND METHODSA reliable and valid Chinese language version of Kogan's Attitudes toward Older People (KAOP) with 17 matched item pairs (34 items) was completed by 275 medical and nursing students. Descriptive data analysis was employed.

RESULTSParticipants reported slightly to have a more positive attitudes (Mean, 144.30 +/- 17.89) than those reported from studies in other countries (Jordanian: Mean, 110.6 +/- 21.79). Students who were females (Mean, 148.27 +/- 18.97), younger (Mean, 146.01 +/- 18.59) and studying nursing (Mean, 156.86 +/- 17.56) were more likely to have a more positive attitudes than older (Mean, 140.04 +/- 15.31), males (Mean, 140.47 +/- 15.93), studying medicine (Mean, 138.77 +/- 15.04).

DISCUSSIONResults show that students may have greater ambivalence attitudes on 10 out of 17 matched item pairs which provides useful information for faculty involved in aged care curriculum design. Working as a volunteer in the university hospital may increase students' interactions with the elderly and may positively influence attitudes towards the elderly.

Aged ; Attitude of Health Personnel ; Female ; Humans ; Male ; Students, Medical ; psychology ; Students, Nursing ; psychology ; Taiwan

Toxoplasma gondii is an obligate intracellular parasite that stimulates production of high levels of proinflammatory cytokines, which are important for innate immunity. NLRs, i.e., nucleotide-binding oligomerization domain (NOD)-like receptors, play a crucial role as innate immune sensors and form multiprotein complexes called inflammasomes, which mediate caspase-1-dependent processing of pro-IL-1β. To elucidate the role of inflammasome components in T. gondii-infected THP-1 macrophages, we examined inflammasome-related gene expression and mechanisms of inflammasome-regulated cytokine IL-1β secretion. The results revealed a significant upregulation of IL-1β after T. gondii infection. T. gondii infection also upregulated the expression of inflammasome sensors, including NLRP1, NLRP3, NLRC4, NLRP6, NLRP8, NLRP13, AIM2, and NAIP, in a time-dependent manner. The infection also upregulated inflammasome adaptor protein ASC and caspase-1 mRNA levels. From this study, we newly found that T. gondii infection regulates NLRC4, NLRP6, NLRP8, NLRP13, AIM2, and neuronal apoptosis inhibitor protein (NAIP) gene expressions in THP-1 macrophages and that the role of the inflammasome-related genes may be critical for mediating the innate immune responses to T. gondii infection.
Apoptosis ; Cytokines ; Gene Expression ; Immunity, Innate ; Inflammasomes* ; Macrophages* ; Multiprotein Complexes ; Negotiating ; Neurons ; Parasites ; RNA, Messenger ; Toxoplasma* ; Up-Regulation

Trichomonas vaginalis secretes a number of proteases which are suspected to be the cause of pathogenesis; however, little is understood how they manipulate host cells. The mammalian target of rapamycin (mTOR) regulates cell growth, cell proliferation, cell motility, cell survival, protein synthesis, and transcription. We detected various types of metalloproteinases including GP63 protein from T. vaginalis trophozoites, and T. vaginalis GP63 metalloproteinase was confirmed by sequencing and western blot. When SiHa cells were stimulated with live T. vaginalis, T. vaginalis excretory-secretory products (ESP) or T. vaginalis lysate, live T. vaginalis and T. vaginalis ESP induced the mTOR cleavage in both time- and parasite load-dependent manner, but T. vaginalis lysate did not. Pretreatment of T. vaginalis with a metalloproteinase inhibitor, 1,10-phenanthroline, completely disappeared the mTOR cleavage in SiHa cells. Collectively, T. vaginalis metallopeptidase induces host cell mTOR cleavage, which may be related to survival of the parasite.
Blotting, Western ; Cell Line, Tumor ; Epithelial Cells/metabolism/parasitology ; Humans ; Metalloproteases/genetics/*metabolism ; Proteolysis ; Sequence Analysis, DNA ; TOR Serine-Threonine Kinases/*metabolism ; Trichomonas vaginalis/*enzymology/genetics

BACKGROUNDInterleukin (IL)-1beta may effectively decrease introcular pressure (IOP) when administered by subconjunctival injection in normal rabbit. However, IL-1beta is a large molecular agent and an inflammation factor. The aim of this study was to evaluate the penetrability of IL-1beta, and the concentrations of both tumor necrosis factor (TNF)-alpha and IL-6 in the aqueous humor of normal rabbits treated with IL-1beta.

METHODSA total of 170 rabbits were used in the study and were assigned to several different treatment groups as follows: 125 of the rabbits were assigned to two groups. In one group, 33 rabbits were injected subconjunctivally with IL-1beta and 39 were injected with saline alone. In the other group, 27 rabbits were given eye drops containing IL-1beta (400 ng/ml) and 26 were given saline alone. Aqueous humor (AH) was drawn and the concentration of IL-1beta within the fluid measured. The IOP was measured in another six rabbits after administration of eye drops containing IL-1beta (400 ng/ml). A further 20 rabbits were assigned to 3 groups as follows: eight untreated normal controls; six injected subconjunctivally with IL-1beta; and six injected subconjunctivally with saline alone. AH was drawn and the concentration of TNF-alpha in the fluid was measured. Another 19 rabbits were assigned to 3 groups as follows: seven untreated normal controls; and six injected subconjunctivally with IL-1beta; and six injected subconjunctivally with saline alone. AH was drawn and the concentration of IL-6 in the fluid measured. Measurement of cytokine concentration was by radio-immunoassay in all cases.

RESULTSThe IL-1beta concentration in the AH was higher in those animals in which it had been administered subconjunctivally (P < 0.01). The IL-1beta concentration in the AH of the animals given eye drops was almost the same as that in the controls (P > 0.05). The administration of IL-1beta in the form of eye drops had little effect upon IOP reduction. Lower TNF-alpha concentrations were seen in the AH after the subconjunctival administration of IL-1beta, but the concentration of IL-6 was the same as in the normal controls.

CONCLUSIONSIL-1beta shows good corneal penetrability after subconjunctival injection into normal rabbit eyes. The IOP reduction induced by IL-1beta is unlikely be associated with an inflammatory response.

Animals ; Aqueous Humor ; drug effects ; metabolism ; Interleukin-1beta ; pharmacology ; Interleukin-6 ; metabolism ; Rabbits ; Tumor Necrosis Factor-alpha ; metabolism

Ascaris suum eggs are inactivated by composting conditions; however, it is difficult to find functional changes in heat-treated A. suum eggs. Here, unembryonated A. suum eggs were incubated at 20degrees C, 50degrees C, and 70degrees C in vitro, and the gene expression levels related to viability, such as eukaryotic translation initiation factor 4E (IF4E), phosphofructokinase 1 (PFK1), and thioredoxin 1 (TRX1), and to apoptosis, such as apoptosis-inducing factor 1 (AIF1) and cell death protein 6 (CDP6), were evaluated by real-time quantitative RT-PCR. No prominent morphological alterations were noted in the eggs at 20degrees C until day 10. In contrast, the eggs developed rapidly, and embryonated eggs and hatched larvae began to die, starting on day 2 at 50degrees C and day 1 at 70degrees C. At 20degrees C, IF4E, PFK1, and TRX1 mRNA expression was significantly increased from days 2-4; however, AIF1 and CDP6 mRNA expression was not changed significantly. IF4E, PFK1, and TRX1 mRNA expression was markedly decreased from day 2 at 50degrees C and 70degrees C, whereas AIF1 and CDP6 mRNA expression was significantly increased. The expressions of HSP70 and HSP90 were detected for 9-10 days at 20degrees C, for 3-5 days at 50degrees C, and for 2 days at 70degrees C. Taken together, incremental heat increases were associated with the rapid development of A. suum eggs, decreased expression of genes related to viability, and earlier expression of apoptosis-related genes, and finally these changes of viability- and apoptosis-related genes of A. suum eggs were associated with survival of the eggs under temperature stress.
Animals ; *Apoptosis ; Ascaris suum/*genetics/*radiation effects ; Cell Survival/radiation effects ; Eggs/radiation effects ; Female ; Gene Expression Profiling ; Gene Expression Regulation/*radiation effects ; Real-Time Polymerase Chain Reaction ; Survival Analysis ; Temperature

BACKGROUNDBecause of the complexity and severity of the surgery and its associated complications, pancreaticoduodenectomy (PD) is associated with significant morbidity and mortality, especially the hemorrhage post-PD. Exploring the factors associated with post-PD hemorrhage is very important for the patients' safety.

METHODSClinical data from 303 cases of PD between January 1998 and December 2008 were analyzed retrospectively.

RESULTSThe overall mortality rate was 4.95% (15/303). However, post-operative bleeding occurred in 25 patients (8.25%) with nine episodes resulting in death (36.00%). Univariate analysis was performed and identified tumor size, Child's classification, total pancreatic uncinatic process resection, and pancreatic leakage as significant risk factors for post-PD hemorrhage. In the severe hemorrhage group, incomplete resection of uncinate process of pancreas and pancreatic leakage were the main causes. The multivariate Logistic regression analysis revealed that each of these variables is an independent risk factor.

CONCLUSIONSPrimary prevention of bleeding complications depends on total pancreatic uncinatic process resection and meticulous hemostatic techniques during surgery. In addition, several peri-operative factors were found to contribute to post-PD bleeding.

Female ; Humans ; Male ; Middle Aged ; Pancreaticoduodenectomy ; adverse effects ; mortality ; Postoperative Hemorrhage ; etiology ; Risk Factors

Toxoplasma gondii is an apicomplexan zoonotic protozoan parasite that infects most species of warm-blooded animals, including humans. The heavy incidence and severe or lethal damage caused by T. gondii infection clearly indicate a need for the development of an effective vaccine. T. gondii GRA8 is a member of the dense granules protein family and is used as a marker of acute infection. In the present study, we evaluated the protective immunity induced by DNA vaccination based on a recombinant eukaryotic plasmid, pDsRed2-GRA8, against acute toxoplasmosis in mice. BALB/c mice were intramuscularly immunized with the pDsRed2-GRA8 plasmid and then challenged by infection with the highly virulent GFP-RH strain of T. gondii. The specific immune responses and protective efficacy against T. gondii of this vaccine were analyzed by measuring cytokine and serum antibody titers, splenocyte proliferation assays, and the survival times of mice after challenge. Our results showed that mice immunized with pDsRed2-GRA8 demonstrated specific humoral and cellular responses, induced higher IgG antibody titers with predominant IgG2a production; increased levels of IL-10, IL-12 (p70), IFN-γ, TNF-α, and splenocyte proliferation; and prolonged survival times compared to those of control mice. The present study showed that DNA immunization with pDsRed2-GRA8 induced humoral and cellular immune responses, and all immunized mice showed greater Th1-type immune responses and longer survival times than those of control mice. These results indicated that T. gondii GRA8 DNA immunization induces a partial protective effect against acute toxoplasmosis.
Animals ; DNA ; Humans ; Immunity, Cellular ; Immunization ; Immunoglobulin G ; Incidence ; Interleukin-10 ; Interleukin-12 ; Mice ; Parasites ; Plasmids ; Toxoplasma ; Toxoplasmosis ; Vaccination

In a previous study, we reported our discovery of Acanthamoeba contamination in domestic tap water; in that study, we determined that some Acanthamoeba strains harbor endosymbiotic bacteria, via our molecular characterization by mitochondrial DNA restriction fragment length polymorphism (Mt DNA RFLP). Five (29.4%) among 17 Acanthamoeba isolates contained endosymbionts in their cytoplasm, as demonstrated via orcein staining. In order to estimate their pathogenicity, we conducted a genetic characterization of the endosymbionts in Acanthamoeba isolated from domestic tap water via 16S rDNA sequencing. The endosymbionts of Acanthamoeba sp. KA/WP3 and KA/WP4 evidenced the highest level of similarity, at 97% of the recently published 16S rDNA sequence of the bacterium, Candidatus Amoebophilus asiaticus. The endosymbionts of Acanthamoeba sp. KA/WP8 and KA/WP12 shared a 97% sequence similarity with each other, and were also highly similar to Candidatus Odyssella thessalonicensis, a member of the alpha-proteobacteria. The endosymbiont of Acanthamoeba sp. KA/WP9 exhibits a high degree of similarity (85-95%) with genus Methylophilus, which is not yet known to harbor any endosymbionts. This is the first report, to the best of our knowledge, to show that Methylophilus spp. can live in the cytoplasm of Acanthamoeba.
Acanthamoeba/isolation & purification/*microbiology/ultrastructure ; Alphaproteobacteria/classification/genetics/*isolation & purification ; Animals ; Bacteroidetes/classification/genetics/*isolation & purification ; Cluster Analysis ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Fresh Water/*parasitology ; Korea ; Methylophilus/classification/genetics/*isolation & purification ; Microscopy, Electron, Transmission ; Phylogeny ; RNA, Ribosomal, 16S/genetics ; Sequence Analysis, DNA ; *Symbiosis