[Show abstract][Hide abstract] ABSTRACT: Cosmetics are normally composed of various ingredients. Some cosmetic ingredients can act as chemical haptens reacting toward proteins or peptides of human skin and they can provoke an immunologic reaction, called as skin sensitization. This haptenation process is very important step of inducing skin sensitization and evaluating the sensitizing potentials of cosmetic ingredients is very important for consumer safety. Therefore, animal alternative methods focusing on monitoring haptenation potential are undergoing vigorous research. To examined the further usefulness of spectrophotometric methods to monitor reactivity of chemicals toward peptides for cosmetic ingredients. Forty chemicals (25 sensitizers and 15 non-sensitizers) were reacted with 2 synthetic peptides, e.g., the cysteine peptides (Ac-RFAACAA-COOH) with free thiol group and the lysine peptides (Ac-RFAAKAA-COOH) with free amine group. Unreacted peptides can be detected after incubating with 5,5'-dithiobis-2-nitrobenzoic acid or fluorescamine™ as detection reagents for free thiol and amine group, respectively. Chemicals were categorized as sensitizers when they induced more than 10% depletion of cysteine peptides or more than 30% depletion of lysine peptides. The sensitivity, specificity, and accuracy were 80.0%, 86.7% and 82.5%, respectively. These results demonstrate that spectrophotometric methods can be an easy, fast, and high-throughput screening tools predicting the skin sensitization potential of chemical including cosmetic ingredient.
[Show abstract][Hide abstract] ABSTRACT: Using a human corneal cell line (HCE-T cells) and 2 evaluation criteria, we developed a new alternative method to assess the eye irritation potential of chemicals. We exposed HCE-T cells to different concentrations of 38 chemicals for 1h and measured relative cell viability (RCV) as an endpoint at each concentration. Using the RCV values, we calculated the RCV50. We also exposed HCE-T cells to 3 fixed concentrations of the 38 chemicals (5%, 0.5%, and 0.05%) for 1h and measured the RCV at each concentration. Using the RCV values at 5%, 0.5%, and 0.05%, we developed a new criterion for eye irritation potential (total eye irritation score, TEIS) and estimated the ocular irritancy. We then assessed the correlation of the results of RCV50 and TEIS with those of the Draize rabbit eye irritation. Both the RCV50 and TEIS results exhibited good positive correlations (sensitivity: 80.77%, specificity: 83.33%, and accuracy: 81.58% for TEIS; sensitivity: 73.08-76.92%, specificity: 75.00%, and accuracy: 73.68-76.32% for RCV50). We conclude that the new in vitro model using HCE-T cells is a good alternative evaluation model for the prediction of the eye irritation potential of chemicals.
[Show abstract][Hide abstract] ABSTRACT: Through our entire life, oral care products such as toothpaste are used. Thus the safety of oral care products used every day to our mouth is very important. As the previous study in animal tests or clinical trials, surfactant in toothpaste may cause the oral irritation. However, EU cosmetics legislation prohibits animal testing of cosmetics and its ingredient for animal welfare. Therefore the development of alternative in vitro test has been actively performed to replace or reduce using the animal in many areas. However, the way to evaluate oral mucosal toxicity has been done using animal models or clinical trials from now on. Even more, the experiment with human oral 3D tissue or human oral cell line is used recently. The aim of this study is the development of oral mucosal irritation method without using animal for the safety of the oral care product. We developed in vitro test method for oral irritation by using human oral cell line (YD-38 cell) acceptable to toothpaste which contains insoluble material. By the results of this assay, we could discriminate toothpaste with or without irritating substance as same manner in animal studies reported previously. In addition, we confirmed that toothpaste for babies and children toothpaste irritated oral musoca lower than the general adult toothpaste. The present study suggest that this new in vitro method by using human oral cell line (YD-38 cell) could be used for evaluation of oral irritation without using animal.
[Show abstract][Hide abstract] ABSTRACT: Topical retinoids inhibit matrix metalloproteinases and accelerate collagen synthesis, thereby triggering antiaging effects in the skin. However, topical retinoids can cause severe skin reactions, including scaling, erythema, papules, and inflammation. The present study demonstrates that the ethanolic bark extract of Alstonia scholaris R. Br. can significantly inhibit all-trans retinoic acid-induced inflammation in human HaCat keratinocyte cells. Furthermore, two representative retinoid-induced proinflammatory cytokines, monocyte chemoattractant protein-1 and interleukin-8, were significantly suppressed by A. scholaris extract (by 82.1% and 26.3% at 100 ppm, and dose-dependently across the tested concentrations) in vitro. In a cumulative irritation patch test, A. scholaris extract decreased retinol-induced skin irritation, while strengthening the ability of retinoids to inhibit matrix metalloproteinase-1 expression, which is strongly associated with aging effects. These results suggest that A. scholaris is a promising compound that may increase the antiaging function of retinoids while reducing their ability to cause skin irritation.
Evidence-based Complementary and Alternative Medicine 01/2012; 2012(11):190370. DOI:10.1155/2012/190370 · 1.88 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In the present work, we assessed the relationship between alkyl carbon chain length and ocular irritation potentials using the hen's egg test-chorioallantoic membrane (HET-CAM) and bovine corneal opacity and permeability (BCOP) assays using 5 commercial alkyl polyglucoside surfactants with different compositions of alkyl chain lengths (C(6)-C(16)). With HET-CAM, there was a good correlation between the proportion of C(10) alkyl polyglucoside and the eye irritation potential Q score (r(2) = 0.912, p = .011). There were no significant differences between the proportion of C(10) alkyl polyglucoside and corneal opacity in BCOP assays; however, there was a relatively high positive correlation between the proportion of C(10) alkyl carbon chain lengths and corneal permeability (r(2) = 0.736, p = .063).
[Show abstract][Hide abstract] ABSTRACT: The attenuation and immunoenhancing effects of rpoS and phoP Salmonella enterica serovar strain Typhi (Salmonella typhi) mutants have not been compared. Here, three S. typhi deletion mutants (phoP, rpoS, and rpoS-phoP double mutant) are constructed and these mutants are characterized with respect to invasiveness, virulence, and protective immune response compared with wild-type Ty2. It was found that phoP and phoP-rpoS deletion mutants are less invasive to HT-29 cells than the wild-type Ty2 and the rpoS single-deleted strain. The LD(50) of immunized mice was higher for phoP than for rpoS mutants, and the highest for the phoP-rpoS double mutant. In addition, all S. typhi mutants showed an increase in the specific serum IgG levels and T-cell-mediated immunity, and showed equal protection abilities against a wild-type Ty2 challenge after two rounds of immunization in BALB/c mice. It is concluded that phoP genes appear to play a more important role than rpoS genes in both cellular invasion and virulence of S. typhi, but not in immunogenicity in mice. Furthermore, the data indicate that the phoP-rpoS double mutant may show promise as a candidate for an attenuated typhoid vaccine.
[Show abstract][Hide abstract] ABSTRACT: Six-week-old male and female Japanese quails (Coturnix japonica) received two organophosphate pesticides, isazofos and pyraclofos, for a 21-day dietary toxicity test, based on the OECD workshop report. During the treatment period, body weight and food consumption of the quail decreased with exposure to either isazofos or pyraclofos. Using the up-and-down procedure to determine the 50% mortality value, we found that the 21-day LC(50) of isazofos and pyraclofos were 40 and 87 mg/kg body weight, respectively. Ataxia, salivation, diarrhea, ruffled feathers, and convulsions at a dead point were observed with both pesticides. The tips of the villi were necrotic in the high dosage groups of isazofos- and pyraclofos-treated quail. Based on these results, body weight, food consumption, clinical signs, and histopathological findings may be useful parameters for detecting the dietary toxicity associated with isazofos and pyraclofos exposure. In addition, Japanese quail could be an excellent bird model for monitoring the toxicological risks of pesticides in Korea.
[Show abstract][Hide abstract] ABSTRACT: The present study was performed to investigate the protective effects of granulocyte macrophage-colony stimulating factor (GM-CSF) against ulcerative mucositis in hamster buccal pouch. GM-CSF was topically administered to the buccal pouches of hamsters with two different doses of 5 and 20 microg/ml. The treatment of GM-CSF led to rapid healing effects in gross and histopathological findings. It decreased expression of pro-inflammatory cytokine mRNA levels in the mucosal tissue of buccal pouches. Also GM-CSF-treated animals showed high numbers of Ki-67 positive cells in basal cell layer. These results suggest that GM-CSF provided excellent healing effects to ulcerative mucositis in the buccal pouch of hamster.
[Show abstract][Hide abstract] ABSTRACT: Mycoplasma pulmonis and Mycoplasma arthritidis were differentially identified using PCR-restriction fragment length polymorphism (RFLP). A genus-specific sequence of mycoplasma was amplified by PCR and the PCR products were digested with the restriction enzyme SmaI. Each PCR product from the four isolates of M. pulmonis was digested with SmaI into two fragments; however, there was no digestion in the PCR product from M. arthritidis. This method might be useful to differentiate infection of M. pulmonis from that of M. arthritidis.
[Show abstract][Hide abstract] ABSTRACT: The antimicrobial effects of a lactic acid producing bacteria culture condensate mixture (LCCM) were assessed against Salmonella enteritidis. In the presence of LCCM, bacterial growth was assessed in vitro by the measurement of optical density (OD) and viable bacterial counting. At concentrations of 1.25 and 2.5% LCCM, OD values were significantly lower than that of the control broth, and at concentrations of 5 and 10% LCCM, OD values did not increase for the entire period of experiment. At 8 h after incubation, the viable bacterial numbers in 5% and 10% LCCM-containing broths were remarkably lower than that in the control broth. This antimicrobial ability of the LCCM was fundamentally attributed to causing cell death rather than inhibiting growth. Even when the pH of LCCM-containing broth was adjusted to 7.2, the number of viable bacteria was significantly lower in the broths containing LCCM over 2.5% than that in control broth at 8 h after incubation. However, the OD value of each culture in the presence of each concentration of the LCCM increased over 1.0 at the completion of the experiment. The in vivo antimicrobial effects of the LCCM against S. enteritidis were also assessed. In S. enteritidis-infected mice, the LCCM decreased both the viable bacteria found in the feces and the mortality rate of the mice. These findings showed that the LCCM might have an antimicrobial ability against S. enteritidis.
International Journal of Food Microbiology 06/2005; 101(1):111-7. DOI:10.1016/j.ijfoodmicro.2004.11.005 · 3.08 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: We report idiopathic intranuclear inclusion bodies in the renal tubular epithelia of two cases of among the 960 Japanese quail (Coturnix coturnix japonica) in the course of the acute oral toxicity and dietary toxicity test. Basophilic inclusion bodies were seen only in the nuclei of renal tubular epithelia. We could not classify our case into any adenovirus infection by clinical signs and lesions. The inclusion bodies were only identified as adenovirus-like particles based upon the electronmicroscopical features.
Journal of Veterinary Science 04/2005; 6(1):75-6. · 1.16 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The immunoenhancing effects of Lactobacillus fermentum PL9005 were assessed via mouse intragastric inoculation. The number of immunoglobulin A-positive cells in the small intestine, CD4+ T lymphocytes in the peripheral blood, and the lymphocyte proliferation response to mitogen stimulation (lipopolysaccharide) increased in mice fed L. fermentum PL9005. The lactic acid concentration also increased dose dependently in the small intestine of mice fed L. fermentum PL9005. No differences were found in body weight, food intake, and clinical signs between mice fed L. fermentum PL9005 and the control group. Results indicated that L. fermentum PL9005 is a probiotic with immunoenhancing properties.
Journal of food protection 04/2005; 68(3):571-6. · 1.85 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This study examined the prevalence of Helicobacter infection in the pyloric mucosa of pigs and its histopathological and molecular characteristics. Forty porcine pyloric samples were examined for Helicobacter infection by silver staining and PCR assay. The PCR product (376 bp) was digested with NdeII to differentiate between Helicobacter heilmannii and Helicobacter pylori. Another PCR assay run to produce an 1157 bp fragment was performed using a primer set designed from the 16S rRNA gene of Candidatus H. suis, and its product was cloned and sequenced. Infection rates were 62.5% (25/40) and 95.0% (38/40) as determined by silver staining and the PCR assay, respectively. On histopathological examination, lymphoid follicle aggregation in the pyloric mucosa and granulocytic migration into the lumen of pyloric glands were observed in 24 (60.0%) and 33 (82.5%) gastric samples, respectively. All PCR products, except that of H. pylori, were cut into two fragments of 147 and 229 bp by enzymatic digestion with NdeII. Sequencing of the 16S rRNA gene showed that the bacterium had 99.57% (1152 bp/1157 bp) homology to the 16S rRNA gene of Candidatus H. suis.
[Show abstract][Hide abstract] ABSTRACT: Epitheliocystis in the carp of a pet fish market were investigated by our diagnostic work and collecting information from department of laboratory animal medicine and fish & shellfish laboratory. The epitheliocystis was identified by using histopathological examination. Epitheliocystis was confirmed as inflammation, epithelial hyperplasia, and lamellar fusion of the gill tissue. Electron microscopic observation showed that the inclusions were filled with Chlamydia-like organism.
Journal of Veterinary Medical Science 01/2005; 67(1):119-20. DOI:10.1292/jvms.67.119 · 0.78 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The efficacy of STEL water for protection against white spot syndrome virus (WSSV) infection was evaluated using shrimp. The LC50 of residual chlorine (Cl-) in STEL water for brood-stock and 2-mo-old shrimp were 2.3 and 3.2 ppm, respectively. All 2-month-old shrimp raised in seawater containing more than 40 microl 2l(-1) of a WSSV-infected tissue homogenate died within 3 d post-exposure (dpe). Thus, a 10-fold dose of 400 microl 2 l(-1) was used in the disinfection tests. Low concentrations of STEL water effectively prevented mortality of shrimp at this challenge dose. All 2-month-old shrimp exposed to seawater with 400 microl of viral homogenate disinfected with STEL water at Cl- concentrations over 0.125 ppm for 1 and 10 min, lived until 5 dpe. With 5-mo-old shrimp, all positive control shrimps died within 3 dpe, whereas most shrimp reared in seawater disinfected with STEL water for 1 h before addition of homogenate lived until 5 dpe. Results suggested that continuous disinfection of seawater with STEL water may be effective for preventing WSSV infection in shrimp.
[Show abstract][Hide abstract] ABSTRACT: Listeria monocytogenes induces the suppurative gastritis in some mice strains. In this study, characteristics of the gastritis caused by L. monocytogenes infection in mice were examined with time course of infection. Mice were administered intragastrically with 1.8 x 10(8) CFU of L. monocytogenes. Each three mice were sacrificed by cervical dislocation at 1, 3, 5, 7, 10, 14, 17, 21, and 28 days postinoculation (pi), respectively. Bacterial colonization in the stomachs reached the peak at 3 days pi, maintained over 4.3 log10 CFU/g tissue until 14 days pi, and was cleared by 28 days pi. However, in the spleens and livers, the bacteria could not be detected after 7 days pi. The gastric lesions were the most prominent at between 3 and 7 days pi. The lesions consisted of marked neutrophilic infiltration, edema, vacuolar degeneration and necrosis of muscle cells and were more severe in the nonglandular region and fundus than in the pylorus, and were in submucosa, lamina muscularis, and serosa than in mucosa. mRNA expression of several cytokines (INF-gamma, IL-1beta, IL-5, IL-6, IL-12, and TNF-alpha) and chemokines (KC, MCP-1) increased in the gastric tissue of infected mice at 1-7 days pi and slightly decreased at 14 days pi. These findings would be useful for studying the pathological mechanism of human febrile gastroenteritis due to L. monocytogenes infection.
[Show abstract][Hide abstract] ABSTRACT: The cholesterol lowering effect of SG-GN3, the extract of salted and fermented small shrimps, Acetes japonicus, was investigated in hypercholesterolemic animal models. Hypercholesterolemia was induced with Triton WR-1339 (nonionic detergent) or high cholesterol (HC)-diet. SG-GN3 significantly decreased total cholesterol (TC) in Triton WR-1339 model at 30 post-treatment hour (549.80 +/- 152.46 mg/dl) compared to the control which induced by only Triton WR-1339 (798.84 +/- 94.98 mg/dl), whereas high-density lipoprotein (HDL) content did not decrease (P < 0.05). In HC-diet model, TC content significantly decreased by SG-GN3 treatment at 3 post-treatment day (P < 0.05). These results suggest that SG-GN3 effectively decreased serum TC level in hypercholesterolemic animal models.
Journal of Ethnopharmacology 05/2004; 91(2-3):231-5. DOI:10.1016/j.jep.2003.12.032 · 3.00 Impact Factor