OBJECTIVE: To investigate the effects of different concentrations of thiomersal on apoptosis and autophagy regulation of human leukemia cell lines U937, CEM-C1 and BALL-1. METHODS: The inhibitory effect of thiomersal on the proliferation of U937, CEM-C1 and BALL-1 cells was detected by CCK-8 assay. Annexin V-FITC/PI double staining flow cytometry was used to detect the apoptosis rate. Western blot was used to detect the effects of thiomersal on autophagy signaling pathway and the expression of PI3K, Akt, mTOR, p-mTOR, caspase-3 and LC3-II proteins. RESULTS: Within 24 and 48 hours, thiomersal inhibited the proliferation of U937, CEM-C1 and BALL-1 cell lines in a time and dose-dependent manner (r_{24 h}=0.295, r_{24 h}=0.452, r_{24 h}=0.103; r_{48 h}=0.821, r_{48 h}=0.665, r_{48 h}=0.821), but no significant time and dose-dependent effect was observed at 72 hours. After 48 hours treatment of thiomersal, the apoptosis rate of U937, CEM-C1 and BALL-1 cells increased in a dose-dependent manner (r=0.819, r=0.763, r=0.835). After 48 hours treatment of thiomersal, the expression levels of PI3K, Akt, mTOR and p-mTOR protein in U937, CEM-C1 and BALL-1 cells decreased in a concentration-dependent manner, the R value of U937 cells was -0.975, -0.899, -0.925 and -0.915, respectively, that of CEM-C1 cells was -0.960, -0.920, -0.861 and -0.927, and that of BALL-1 cells was -0.939, -0.911, -0.896 and -0.926,. which suggested that thiomersal-induced apoptosis of U937, CEM-C1 and BALL-1 cells might be due to the inhibition of PI3K/Akt/mTOR pathway. Thiomersal promoted the apoptosis of U937, CEM-C1 and BALL-1 cells via caspase-3 pathway, and the expressions of caspase-3 and LC3-II were up-regulated in a dose-dependent manner (r=0.976, r=0.914; r=0.976, r=0.986; r=0.961, r=0.974). CONCLUSIONS Thiomersal can inhibit the proliferation and promote the apoptosis of U937, CEM-C1 and BALL-1 cells. A certain concentration of thiomersal can down-regulate the expression of PI3K/Akt/mTOR pathway related proteins PI3K, Akt, mTOR and p-mTOR in U937, CEM-C1 and BALL-1 cells, and activate autophagy and apoptosis by down-regulation of PI3K/Akt/mTOR pathway.
Humans ; Thimerosal ; Caspase 3 ; Phosphatidylinositol 3-Kinases ; Autophagy ; Apoptosis ; Leukemia ; Cell Line

BACKGROUND: Rosacea is a relatively common inflammatory skin disease characterized by central facial erythema that persists for several months or longer. Experimental studies have demonstrated that rosacea compromises the epidermal barrier protection against allergens and irritants, rendering rosacea patients susceptible to contact hypersensitivity and skin irritation. OBJECTIVE: This study aimed to investigate allergic and irritant reactions to topical agents and cosmetics in rosacea patients using patch tests. METHODS: Rosacea signs and subtypes of 40 patients were recorded and subjective symptoms and clinical experiences related to topical agents and cosmetics were assessed using a questionnaire. Patch tests were performed in these patients using the Korean Standard Series (Chemotechnique Diagnostics, Sweden), the TRUE test® (Mekos Laboratories ApS, Denmark), and 26 selected fragrance and 31 cosmetic antigens (TROLAB, Germany). RESULTS: Of 40 patch-tested patients, 26 (65%) showed at least 1 positive reaction and 13 (32.5%) showed 2 or more positive reactions. The most common allergens were nickel (II) sulfate (50%), cobalt chloride (10%), epoxy resin (7.7%), and thimerosal (7.5%). Thirteen patients (32.5%) experienced irritant patch test reactions. There were no significant differences in allergic or irritant positive reactions between mild, moderate, and severe cases of rosacea or between rosacea subtypes. Irritant reactions were more common in fair-skinned participants. CONCLUSION: Different topical medications are used in the treatment of rosacea patients. Exacerbation of rosacea may be caused by a hypersensitivity reaction or irritation induced by the topical agent or by cosmetics.
Allergens ; Clinical Study ; Cobalt ; Dermatitis, Contact ; Erythema ; Humans ; Hypersensitivity ; Irritants ; Nickel ; Patch Tests ; Rosacea ; Skin ; Skin Diseases ; Thimerosal

PURPOSE: An oral cholera vaccine (OCV), Euvichol, with thimerosal (TM) as preservative, was prequalified by the World Health Organization (WHO) in 2015. In recent years, public health services and regulatory bodies recommended to eliminate TM in vaccines due to theoretical safety concerns. In this study, we examined whether TM-free Euvichol induces comparable immunogenicity to its TM-containing formulation in animal model. MATERIALS AND METHODS: To evaluate and compare the immunogenicity of the two variations of OCV, mice were immunized with TM-free or TM-containing Euvichol twice at 2-week interval by intranasal or oral route. One week after the last immunization, mice were challenged with Vibrio cholerae O1 and daily monitored to examine the protective immunity against cholera infection. In addition, serum samples were obtained from mice to measure vibriocidal activity and vaccine-specific IgG, IgM, and IgA antibodies using vibriocidal assay and enzyme-linked immunosorbent assay, respectively. RESULTS: No significant difference in immunogenicity, including vibriocidal activity and vaccine-specific IgG, IgM, and IgA in serum, was observed between mice groups administered with TM-free and -containing Euvichol, regardless of immunization route. However, intranasally immunized mice elicited higher levels of serum antibodies than those immunized via oral route. Moreover, intranasal immunization completely protected mice against V. cholerae challenge but not oral immunization. There was no significant difference in protection between two Euvichol variations. CONCLUSION: These results suggested that TM-free Euvichol could provide comparable immunogenicity to the WHO prequalified Euvichol containing TM as it was later confirmed in a clinical study. The pulmonary mouse cholera model can be considered useful to examine in vivo the potency of OCVs.
Animals* ; Antibodies ; Cholera Vaccines ; Cholera* ; Clinical Study ; Enzyme-Linked Immunosorbent Assay ; Immunization ; Immunoglobulin A ; Immunoglobulin G ; Immunoglobulin M ; Mice ; Models, Animal* ; Public Health ; Thimerosal ; Vaccines ; Vibrio cholerae O1 ; World Health Organization

OBJECTIVE: This study was to identify small inhibitory RNAs (siRNAs) that are effective in inhibiting growth of cervical cancer cell lines harboring human papilloma virus (HPV) and to examine how siRNAs interact with interferon beta (IFN-beta) and thimerosal. METHODS: The HPV18-positive HeLa and C-4I cell lines were used. Four types of siRNAs were designed according to their target (both E6 and E7 vs. E6 only) and sizes (21- vs. 27-nucleotides); Ex-18E6/21, Ex-18E6/27, Sp-18E6/21, and Sp-18E6/27. Each siRNA-transfected cells were cultured with or without IFN-b and thimerosal and their viability was measured. RESULTS: The viabilities of HPV18-positive tumor cells were reduced by 21- and 27-nucleotide siRNAs in proportion to the siRNA concentrations. Of the two types of siRNAs, the 27-nucleotide siRNA constructs showed greater inhibitory efficacy. Sp-18E6 siRNAs, which selectively downregulates E6 protein only, were more effective than the E6- and E7-targeting Ex-18E6 siRNAs. siRNAs and IFN-beta showed the synergistic effect to inhibit HeLa cell survival and the effect was proportional to both siRNA and IFN-beta concentrations. Thimerosal in the presence of siRNA exerted a dose-dependent inhibition of C-4I cell survival. Finally, co-treatment with siRNA, IFN-beta, and thimerosal induced the most profound decrease in the viability of both cell lines. CONCLUSION: Long (27-nucleotides) siRNAs targeting E6-E7 mRNAs effectively reduce the viability of HPV18-positive cervical cancer cells and show the synergistic effect in combination with IFN-b and thimerosal. It is necessary to find the rational design of siRNAs and effective co-factors to eradicate particular cervical cancer.
Cell Line ; Cell Survival ; HeLa Cells ; Humans ; Interferon-beta ; Papilloma* ; RNA* ; RNA, Messenger ; RNA, Small Interfering ; Thimerosal ; Uterine Cervical Neoplasms*

BACKGROUND: A number of skin manifestations in patients with allergic contact dermatitis result from sensitization with specific allergens, and patch testing is used as a diagnostic means of identifying theses allergens. In Korea, the commercial patch test panel, the TRUE-test, has been available since 2005. However, there have been few reports regarding the results of the TRUE-test. OBJECTIVE: The purpose of this study was to analyze the type, frequency, and changes in common contact allergens in Korea using the TRUE test. METHODS: A total of 311 patients with allergic contact dermatitis were evaluated by the TRUE test between January 2007 and December 2011 at Soonchunhyang hospital. Patch test reading was performed on day 2 and day 4. RESULTS: A total of 311 cases of TRUE test records (male 79, female 232) were compiled and analyzed. The highest age distribution was the 4th decade in females. The face, with exception of the lips, was the most frequently affected site (40.2%). Overall, 58.8% of patients had at least on positive reaction, and the most common allergens were nickel sulfate (31.8%), p-phenylenediamine (13.5%), cobalt chloride (10.0%), thiomersal (7.4%), and carba mix (5.5%). Nickel allergen displayed higher positive rates than the rates in other countries. CONCLUSION: There was no significant difference in the overall positive rate of patch test results compared with recent studies. However, we confirmed that metal-related allergens remain the most common, and that the results provide the basis for the use of the TRUE test in patients with contact dermatitis in Korea.
Age Distribution ; Allergens ; Cobalt ; Dermatitis, Allergic Contact ; Dermatitis, Contact ; Ditiocarb ; Female ; Guanidines ; Humans ; Korea ; Lip ; Nickel ; Patch Tests ; Phenylenediamines ; Skin Manifestations ; Thimerosal

Previously, we reviewed biological evidence that mercury could induce autoimmunity and coronary arterial wall relaxation as observed in Kawasaki syndrome (KS) through its effects on calcium signaling, and that inositol 1,4,5-triphosphate 3-kinase C (ITPKC) susceptibility in KS would predispose patients to mercury by increasing Ca2+ release. Hg2+ sensitizes inositol 1,4,5-triphosphate (IP3) receptors at low doses, which release Ca2+ from intracellular stores in the sarcoplasmic reticulum, resulting in delayed, repetitive calcium influx. ITPKC prevents IP3 from triggering IP3 receptors to release calcium by converting IP3 to inositol 1,3,4,5-tetrakisphosphate. Defective IP3 phosphorylation resulting from reduced genetic expressions of ITPKC in KS would promote IP3, which increases Ca2+ release. Hg2+ increases catecholamine levels through the inhibition of S-adenosylmethionine and subsequently catechol-O-methyltransferase (COMT), while a single nucleotide polymorphism of the COMT gene (rs769224) was recently found to be significantly associated with the development of coronary artery lesions in KS. Accumulation of norepinephrine or epinephrine would potentiate Hg2+-induced calcium influx by increasing IP3 production and increasing the permeability of cardiac sarcolemma to Ca2+. Norepinephrine and epinephrine also promote the secretion of atrial natriuretic peptide, a potent vasodilator that suppresses the release of vasoconstrictors. Elevated catecholamine levels can induce hypertension and tachycardia, while increased arterial pressure and a rapid heart rate would promote arterial vasodilation and subsequent fatal thromboses, particularly in tandem. Genetic risk factors may explain why only a susceptible subset of children develops KS although mercury exposure from methylmercury in fish or thimerosal in pediatric vaccines is nearly ubiquitous. During the infantile acrodynia epidemic, only 1 in 500 children developed acrodynia whereas mercury exposure was very common due to the use of teething powders. This hypothesis mirrors the leading theory for KS in which a widespread infection only induces KS in susceptible children. Acrodynia can mimic the clinical picture of KS, leading to its inclusion in the differential diagnosis for KS. Catecholamine levels are often elevated in acrodynia and may also play a role in KS. We conclude that KS may be the acute febrile form of acrodynia.
Acrodynia ; Arterial Pressure ; Autoimmunity ; Calcium ; Calcium Signaling ; Catechol O-Methyltransferase ; Catecholamines ; Child ; Coronary Vessels ; Diagnosis, Differential ; Epinephrine ; Heart Rate ; Humans ; Hydrazines ; Hypertension ; Inositol ; Inositol 1,4,5-Trisphosphate ; Inositol 1,4,5-Trisphosphate Receptors ; Inositol Phosphates ; Mucocutaneous Lymph Node Syndrome ; Norepinephrine ; Permeability ; Phosphorylation ; Polymorphism, Single Nucleotide ; Powders ; Relaxation ; Risk Factors ; S-Adenosylmethionine ; Sarcolemma ; Sarcoplasmic Reticulum ; Tachycardia ; Thimerosal ; Thrombosis ; Tooth ; Tooth Eruption ; Vaccines ; Vasoconstrictor Agents ; Vasodilation

BACKGROUND: Patch test is usually performed in order to differentiate allergic contact dermatitis from other facial skin diseases; however, there are only infrequent comparative studies regarding the positive allergens patch tested between allergic contact dermatitis and other facial skin diseases. OBJECTIVE: The purpose of this study was to investigate the positivity of patch test in patients with facial dermatitis and to analyze the valid positive allergen patch tested in allergic contact dermatitis compared to other facial skin diseases. In addition, we evaluated the difference of positive allergen according to clinical features, such as age, sex, site of the lesion and duration of disease, in patients with allergic contact dermatitis. METHODS: A total of 409 patients tested with facial dermatitis patch were retrospectively reviewed for medical recordings, including clinical features, occupational history, clinical photos and the results of a Korean standard series of patch tests. RESULTS: Compared with other facial skin disease, patients diagnosed with allergic contact dermatitis showed higher rate in the patch test, responding to 2 or more allergens. They also had more statistically valid positive allergens, such as phenylendiamine base, cobalt, colophony, thimerosal and isothizolinone. In addition, there were a few allergens which revealed significant differences in the positivity of the patch test according to the clinical features in patients diagnosed with allergic contact dermatitis. CONCLUSION: Based on these results, we could receive help in order to classify the positive allergens as relevant allergens in the patch test for the differentiation of allergic contact dermatitis from other facial skin disease.
Allergens ; Cobalt ; Dermatitis ; Dermatitis, Allergic Contact ; Humans ; Medical Records ; Patch Tests ; Resins, Plant ; Retrospective Studies ; Skin ; Skin Diseases ; Thimerosal

Active sensitization of contact allergen is an uncommon complication of patch test. Para-tertiary-butylphenol (PTBP) is a component of neoprene-based adhesives used in glued leather goods such as shoes, handbags, belts and watchstraps. Contact allergy to PTBP is frequently seen when the patch test is performed. However, active sensitization to PTBP after the patch test is rare. A 23-year-old female patient came to our clinic with a brownish hyperpigmented patch on the belt position of her belly. Pruritic erythematous papules and vesicles developed on the abdomen when she wore new pants 4 months ago. Under the clinical diagnosis of allergic contact dermatitis, the patient was patch tested with the Thin-layer Rapid Use Epicutaneous Test (TRUE Test(R)). The result showed strong positive reaction to nickel sulphate and thiomersal both on day 2 and 3 after application. Twenty-nine days after the test, the patient noticed pruritus on the TRUE Test(R) area. Thirty-four days after the test, an erythematous square-shaped plaque studded with tiny vesicles developed at the PTBP patch test site. This strongly suggests that the reaction to PTBP is occurred. She was diagnosed with an active sensitization to PTBP. We report a rare and interesting case of active sensitization to PTBP after TRUE Test(R).
Abdomen ; Adhesives ; Dermatitis, Allergic Contact ; Dermatitis, Contact ; Female ; Humans ; Hypersensitivity ; Nickel ; Patch Tests ; Phenols ; Porphyrins ; Pruritus ; Shoes ; Thimerosal

BACKGROUND: The patch test is widely used for diagnosis of allergic contact dermatitis. However, nearly half of positive reactions can be observed only on day 2 or day 4 and it is difficult to interpret these reactions. OBJECTIVE: The purpose of this study is to assess the frequency of transient and delayed reactions in TRUE-test and detect common antigens that provoke these reactions. METHODS: A total of 311 patients with allergic contact dermatitis were evaluated by TRUE-test between Jan 2007 and December 2011. Records of patch test results of day 2 and day 4 were reviewed and analyzed. RESULTS: A total 311 cases of T.R.U.E. TEST(R) records (male 79, female 232) were analyzed. Persistent reactions were observed in 80.1% patients tested, transient reactions were observed in 18.3%, and delayed reaction in 5%. Frequent allergens which showed transient reactions were cobalt dichloride (2.9%), nickel sulfate (2.2%), thiomersal (1.9%), and carba mix (1.6%), in order of frequency. Allergens which showed delayed reactions were nickel sulfate (0.3%), fragrance mix (0.3%), p-tert-butylphenol formaldehyde resin (0.43). CONCLUSION: Our results showed a relatively high frequency of transient reaction in T.R.U.E. TEST(R). This suggests that additional reading at day 4 in the patch test would be of value.
Allergens ; Cobalt ; Dermatitis, Allergic Contact ; Ditiocarb ; Female ; Formaldehyde ; Guanidines ; Humans ; Nickel ; Patch Tests ; Resins, Synthetic ; Thimerosal

BACKGROUND: Allergic contact dermatitis (ACD) was once regarded as a disorder of the adult population, while ACD in children was considered rare. However, ACD in children may be more common than previously realized and more recently, it has been estimated that more than 20% of the pediatric population is affected by ACD. However, in Korea results of patch testing in the pediatric population has not yet been reported. OBJECTIVE: The purpose of this study was to analyze allergens responsible for ACD in Korean children and investigate the influence of sex, involved site, and atopic dermatitis on contact allergen sensitization. We also wanted to compare the results between the pediatric and the adult group. METHODS: A retrospective analysis of all patch test results from our data base between 2009 and 2011 was performed. A total of 234 patients were patch tested. The pediatric population was defined as patients 18 years and younger, and total 30 pediatric patients were included. RESULTS: A total of 30 patch tested cases (male 12, female 18) were analyzed. Overall, 66.7% of pediatric patients had at least one positive reaction and common allergens were nickel sulfate (33.3%), thimerosal (13.0%), and black rubber mix (10.0%) in order of frequency. There were no significant differences between contact allergen sensitivity and sex or involved sites. However, in the atopic group, the positive reactions to wool alcohols were significantly higher than in the non-atopic group (p=0.0076). In adults, common allergens were nickel sulfate (34.8%), p-tert butylphenol formaldehyde resin (11.8%), cobalt chloride (11.3%) and thimerosal (11.3%) in order of frequency. This was not significantly different to the pediatric group. CONCLUSION: ACD in children is not uncommon and patch testing in suspected children revealed 66.7% of positive reaction.
2-Naphthylamine ; Adult ; Alcohols ; Allergens ; Child ; Cobalt ; Dermatitis, Allergic Contact ; Dermatitis, Atopic ; Female ; Formaldehyde ; Humans ; Korea ; Nickel ; Patch Tests ; Pediatrics ; Phenylenediamines ; Retrospective Studies ; Rubber ; Thimerosal ; Wool