The aim of this study was to identify two homologous serine proteinase inhibitor (serpin) molecules, squamous cell carcinoma (SCC) antigen-1 and -2, by two-dimensional electrophoresis (2-DE), combined with immunoblotting, and examine their expression in tumor tissue. The recombinant SCC (rSCC) antigen-1 showed four spots with p/ 6.5, 6.4, 6.3 and 6.0, whereas rSCC antigen-2 showed a more acidic spot with p/5.95. SCC antigen in tumor tissue appeared in three new acidic spots (p/5.7-5.5, M(r) 44 500), numbered 5, 6 and 7, besides the previously reported four spots numbered 1 to 4. These new acidic spots of SCC antigen apparently increased in SCC tissue. Treatment of tissue extract by carboxymethyl (CM)-papain agarose matrix extinguished spots 1 to 4 encoded on the SCCA1 gene, but not 5 to 7 on the SCCA2 gene. Overexpression of the SCCA2 gene may play an important role in the malignant behavior of tumor cells.
A divalent thiaruthenacycle complex, cis-Ru[SC6H3(2-CH2)(6-Me)-κ2S,C](PMe3)4 (3), is prepared by the treatment of Ru(η4-1,5-COD)(η6-1,3,5-COT) (1) with 2,6-dimethylbenzenethiol in the presence of PMe3 via Ru(η5-cyclooctadienyl)(SC6H3Me2-2,6)(PMe3)2 (2). Exposure of 3 in benzene to H2 (0.1 MPa) leads to the quantitative formation of cis-RuH(SC6H3Me2-2,6)(PMe3)4 (4), which readily turns to 3 at room temperature on evacuation, indicating the reversibility of the reaction. Both forward and backward reactions of this equilibrium are retarded by addition of PMe3, suggesting prerequisite prior dissociation of PMe3 for both reactions. Complex 3 catalyzes selective and facile deuteration of the ortho-methyl and the mercapto groups in 2,6-dimethylbenzenethiol under D2.
Squamous cell carcinoma antigen (SCCA) is expressed in normal squamous epithelia and malignant squamous cell tissues. The serum level of SCCA has been used to evaluate treatment efficacy, clinical course of disease, and recurrence. SCCA is produced by at least two genes (SCCA1 and SCCA2); both of them have been located on chromosome 18q21.3. It has been difficult to examine the expression levels of SCCA1 and SCCA2 mRNAs separately because of their high homology at nucleotide level. In the present study, asymmetric semi-nested reverse transcription PCR, based on the principle of fluorescence energy transfer, enabled to quantitate the copy numbers of both SCCA1 and SCCA2 mRNAs. Using this method, the expression levels of these mRNAs were evaluated in normal and malignant squamous tissues. The copy number of SCCA2 mRNA was higher in malignant tissues than in normal tissues, while those of SCCA1 mRNA did not significantly differ between normal and malignant tissues. These data indicate that specific quantitation of the expression level of SCCA2 mRNA may be useful for the diagnosis and management of patients with squamous cell carcinoma.
Abstract The reactions of 4‐bromoacetyl‐3‐methoxy‐3‐methyl‐6,6‐diphenyl‐1,2‐dioxane with thioureas or thioamides gave 3‐methoxy‐3‐methyl‐6,6‐diphenyl‐4‐(4‐thiazolyl)‐1,2‐dioxanes in 63–90% yields. The similar reaction of 4‐bromoacetyl‐3‐methoxy‐3‐methyl‐6,6‐diphenyl‐1,2‐dioxane with acetamide gave 3‐methoxy‐3‐methyl‐4‐(2‐methyl‐4‐oxazolyl)‐6,6‐diphenyl‐1,2‐dioxane in 39% yields. The reactions of 4‐bromoacetyl‐3‐methoxy‐3‐methyl‐6,6‐diphenyl‐1,2‐dioxane with 3‐alkyl‐4‐amino‐5‐mercaptot[1,2,4]triazoles yielded 3‐methoxy‐3‐methyl‐6,6‐diphenyl‐4‐[3‐(5‐alkyl[1,2,4]triazolo[3,4‐ b ]‐2,3‐dihydro‐6 H ‐[1,3,4]thiadiazinyl)]‐1,2‐dioxanes in moderate yields (43–46%).
DNA vaccination can be applied to the treatment of various infectious diseases and cancers; however, technical difficulties have hindered the development of an effective delivery method. The efficacy of a DNA vaccine depends on optimal antigen expression by the injected plasmid DNA. The pyro-drive jet injector (PJI) is a novel system that allows for adjustment of injection depth and may, thus, provide a targeted delivery approach for various therapeutic or preventative compounds. Herein, we investigated its potential for use in delivering DNA vaccines. This study evaluated the optimal ignition powder dosage, as well as its delivery effectiveness in both rat and mouse models, while comparing the results of the PJI with that of a needle syringe delivery system. We found that the PJI effectively delivered plasmid DNA to intradermal regions in both rats and mice. Further, it efficiently transfected plasmid DNA directly into the nuclei, resulting in higher protein expression than that achieved via needle syringe injection. Moreover, results from animal ovalbumin (OVA) antigen induction models revealed that animals receiving OVA expression plasmids (pOVA) via PJI exhibited dose-dependent (10 μg, 60 μg, and 120 μg) production of anti-OVA antibodies; while only low titers (< 1/100) of OVA antibodies were detected when 120 μg of pOVA was injected via needle syringe. Thus, PJI is an effective, novel method for delivery of plasmid DNA into epidermal and dermal cells suggesting its promise as a tool for DNA vaccination.
Neonicotinoids are potent agonists of nicotinic acetylcholine receptors that exert insecticidal effects by causing abnormal excitation of the nervous system. Neonicotinoids and their metabolites effect in mammals, including humans, have become a concern. In the present study, we evaluated the effects of chronic exposure of two neonicotinoids, acetamiprid (ACE) and clothianidin (CTD), on Caenorhabditis elegans. We used 1, 10, 100, and 1000 µM solutions of nicotine, ACE, and CTD dissolved in 1% dimethyl sulfoxide (DMSO). Bioassays and motility tests, which are neurotoxicity assessments, were performed on the L1‒L2 larvae of wild-type C. elegans. To evaluate the effect of exposure over multiple generations and the correlation between concentrations and generations, the same study was conducted on the second and third generations of the exposed group. The bioassay results showed concentration-dependent adverse effects: body length, maturity rate, and lifetime number of pups decreased for both ACE and CTD for the first generation. In a multi-generation study, the effect intensified with the progression of generations, and the toxicity of both ACE and CTD was cumulative. This effect was more pronounced in breeding studies. The motility test results showed concentration-dependent adverse effects, such as a decrease in the number of behaviors for both ACE and CTD in both tests for the first generation. In a multi-generation study, the effect intensified with the progression of generations, and this effect was more pronounced with ACE exposure. Thus, the chronic exposure to ACE and CTD may cause cross-generational adverse effects, especially on C. elegans reproduction and motion.
A single reference high-performance liquid chromatographic (SR-HPLC) method was developed and validated for the therapeutic drug monitoring (TDM) of phenytoin (PHT) and carbamazepine (CBZ) in plasma from patients. The analytical parameters evaluated were linearity, limit of quantification (LOQ), selectivity, accuracy, and stability according to the US Food and Drug Administration (FDA) guideline. The developed method shows good linearity (r2 > 0.999; LOQ—50 µg/mL), and LOQ values were 1.56 µg/mL for PHT and 0.40 µg/mL for CBZ at 254 nm. For the development of SR-HPLC method, we evaluated to improve the detection wavelength, stirred retention time, and stability for SR, and selected 5-(p-methylphenyl)-5-phenylhydantoin for PHT (relative molar sensitivity, RMS = 0.848) and 10-methoxyiminostilbene for CBZ (RMS = 0.263). The established differential definite quantities of PHT and CBZ in plasma samples were similar using the RMS and absolute calibration methods based on RSD < 5.10%. A preliminary application was performed using chemiluminescent immunoassay and SR-HPLC method, in which the detectable values of the correlation coefficient and the slope of the intercept were PHT: 0.964 and 0.992647, and CBZ: 0.969 and 1.072089, respectively. Based on these results, we propose that the SR-HPLC method with RMS would be offered to the useful and accurate TDM of various medicines in plasma/serum samples.
In recent years, paraoxybenzoic acid esters (parabens) have been used in pharmaceuticals, cosmetics, and food additives. Parabens have been reported to have a weak estrogenic effect in in vitro test systems, and it is presumed that the longer the alkyl chain of the paraben, the greater its endocrine-disrupting and reproductive function effects. However, the effects of parabens on human health are still unclear. In this study, we evaluated the effects of six parabens (methyl p-hydroxybenzoate [MP], ethyl p-hydroxybenzoate [EP], propyl p-hydroxybenzoate [PP], isopropyl p-hydroxybenzoate [IPP], butyl p-hydroxybenzoate [BP], and isobutyl p-hydroxybenzoate [IBP]) on the reproductive function of the model organism Caenorhabditis elegans. We used 25, 50, and 100 µg/mL solutions of parabens in 0.1% dimethyl sulfoxide (DMSO). Bioassays (growth and maturation effect tests and reproduction effect tests) were performed on L1 larvae of wild-type C. elegans. In the growth effects test, all parabens were observed to have no effect. In the maturation effects test, there was a significant decrease in maturity at each concentration of five of the six parabens, with the exception being MP. In the reproduction effects test, a significant decrease in the number of lifetime offspring was observed at each concentration of five of the six parabens, with the exception being EP. This decrease was remarkable with PP, which has been reported to adversely affect reproductive function in rats. It is necessary to continue to focus on the estrogen-like action of parabens, including PP, and perform genetic analyses, such as RNA sequencing.
