Intramuscular electroporation with the pro-opiomelanocortin gene in rat adjuvant arthritis

Endogenous opioid peptides have an essential role in the intrinsic modulation and control of inflammatory pain, which could be therapeutically useful. In this study, we established a muscular electroporation method for the gene transfer of pro-opiomelanocortin (POMC) in vivo and investigated its effect on inflammatory pain in a rat model of rheumatoid arthritis. The gene encoding human POMC was inserted into a modified pCMV plasmid, and 0-200 microg of the plasmid-POMC DNA construct was transferred into the tibialis anterior muscle of rats treated with complete Freund's adjuvant (CFA) with or without POMC gene transfer by the electroporation method. The safety and efficiency of the gene transfer was assessed with the following parameters: thermal hyperalgesia, serum adrenocorticotropic hormone (ACTH) and endorphin levels, paw swelling and muscle endorphin levels at 1, 2 and 3 weeks after electroporation. Serum ACTH and endorphin levels of the group into which the gene encoding POMC had been transferred were increased to about 13-14-fold those of the normal control. These levels peaked 1 week after electroporation and significantly decreased 2 weeks after electroporation. Rats that had received the gene encoding POMC had less thermal hypersensitivity and paw swelling than the non-gene-transferred group at days 3, 5 and 7 after injection with CFA. Our promising results showed that transfer of the gene encoding POMC by electroporation is a new and effective method for its expression in vivo, and the analgesic effects of POMC cDNA with electroporation in a rat model of rheumatoid arthritis are reversed by naloxone.     

Synthesis,in vitro structure–activity relationship,and in vivo studies of 2-arylthiazolidine-4-carboxylic acid amides as anticancer agents

A series of (2RS,4R)-2-arylthiazolidine-4-carboxylic acid amide (ATCAA) was synthesized. Antiproliferative activity against melanoma and prostate cancer cells compared with control cells (fibroblast and RH7777, respectively) was evaluated. Compound 3id showed the best selectivity and growth-inhibition activity against three melanoma cell lines (B16-F1, A375, and WM-164). Compounds 15b and 3ac had good selectivity and potency against four prostate cancer cell lines (DU 145, PC-3, LNCaP, and PPC-1). The structure–activity relationship (SAR) of the side chain, the thiazolidine ring, and phenyl substituents is discussed. Cell cycle analysis showed that the percentage of cancer cells undergoing apoptosis (sub-G1 phase) increased after treatment with 1b and 3ad, which also strongly inhibited melanoma colony formation. In vivo studies on nude mice bearing A375 melanoma tumors showed that compound 1b inhibited tumor growth in a dose-dependent manner. At a dose of 10 mg/kg, 1b significantly inhibited melanoma tumor growth and showed higher efficacy than did dacarbazine at 60 mg/kg.     

First report of human biting activity of Ixodes acutitarsus (Acari: Ixodidae) collected in Taiwan

Ixodes acutitarsus is said to be the largest Ixodes tick in the world. Human biting activity of I. acutitarsus was reported for the first time in Taiwan. Ticks were observed from April to December and collected from infested humans during trips to the mountainous areas of northern, central, southern and eastern Taiwan. Host range, seasonal prevalence, and the vectorial capacity of I. acutitarsus ticks for various human pathogens need to be further identified.     

Utilization of ground eggshell waste as an adsorbent for the removal of dyes from aqueous solution

The adsorption of cationic basic blue 9 and anionic acid orange 51 from aqueous solution onto the calcified eggshell (ES) and its ground eggshell powder (ESP) was carried out by varying the process parameters such as agitation speed, initial dye concentration, adsorbent mass and temperature. The adsorption potential for basic blue 9 onto ESP is far lower than that for acid orange 51, mainly due to the ionic interaction between the acid dye with the sulfonate groups and the positively charged sites on the surface of ESP. The adsorption capacity of acid orange 51 onto ES is significantly smaller than that onto ESP, which is in line with their pore properties (i.e., 1 vs. 21 m(2)/g). The experimental results showed that the adsorption process can be well described with a simple model, the pseudo-second-order model. According to the equilibrium adsorption capacity from the fitting of pseudo-second order reaction model, it was further found that the Freundlich model yields a somewhat better fit than the Langmuir model in the adsorption of acid orange 51 onto ESP. In addition, an increase in adsorption temperature from 15 to 45 degrees C significantly enhances the adsorption capacity of acid orange 51 onto ESP, revealing that the adsorption should be an endothermic or chemisorption process. From the results, it is feasible to utilize the ground eggshell waste as an effective adsorbent for removal of anionic dye from aqueous solution.     

Molecular analysis of <Emphasis Type="Italic">Ixodes granulatus</Emphasis>, a possible vector tick for <Emphasis Type="Italic">Borrelia burgdorferi</Emphasis> sensu lato in Taiwan

The genetic identity of Ixodes granulatus ticks was determined for the first time in Taiwan. The phylogenetic relationships were analyzed by comparing the sequences of mitochondrial 16S ribosomal DNA gene obtained from 19 strains of ticks representing seven species of Ixodes and two outgroup species (Rhipicephalus sanguineus and Haemaphysalis inermis). Four major clades could be easily distinguished by neighbour-joining analysis and were congruent by maximum-parsimony method. All these I. granulatus ticks of Taiwan were genetically affiliated to a monophyletic group with highly homogeneous sequences (92.2–99.3% similarity), and can be discriminated from other Ixodes species and other genera of ticks with a sequence divergence ranging from 11.7 to 30.8%. Moreover, intraspecific analysis revealed that two distinct lineages are evident between the same species of I. granulatus ticks collected from Taiwan and Malaysia. Our results demonstrate that all these I. granulatus ticks of Taiwan represent a unique lineage distinct from the common vector ticks (I. ricinus complex) for Borrelia burgdorferi spirochetes.     

Species identification of <Emphasis Type="Italic">Ixodes granulatus</Emphasis> (Acari: Ixodidae) based on internal transcribed spacer 2 (ITS2) sequences

To investigate the genetic specificity of Ixodes granulatus ticks collected from Taiwan, the genetic identities and phylogenetic relationships were analyzed by comparing the sequences of the internal transcribed spacer 2 (ITS2) region obtained from 27 strains of ticks representing twelve species of Ixodes. Five major clades can be easily distinguished by neighbour-joining analysis and were congruent by maximum-parsimony method. All these I. granulatus ticks collected from Taiwan and Japan were genetically affiliated to a monophyletic group with highly homogeneous sequences (95.8–99.5% similarity), and can be discriminated from other species and subgenera of Ixodes ticks with a sequence divergence ranging from 13.6% to 62.9%. Moreover, interspecific analysis revealed that four distinct lineages are evident between Ixodes ticks, and all these I. granulatus ticks collected from Taiwan and Japan belong to the same lineage. Our results provide the first investigation on the genetic specificity of I. granulatus ticks, and demonstrate that all these I. granulatus ticks represent a unique lineage distinct from other species and subgenera of Ixodes ticks. The feasibility of ITS2-based genetic analysis for species-specific identification of I. granulatus ticks around East Asia was highly anticipated.     

Effect of triton X-100 on alkaline lipase production by Pseudomonas pseudoalcaligenes F-111

Summary An extracellular alkaline lipase was evaluated from Pseudomonas pseudoalcaligenes F-111, which was isolated from soil using a rhodamine B agar plate with Na₂CO₃. For enzyme production, olive oil, soymeal, triton X-100 and sodium ion were found to be essential. The addition of triton X-100 increased the alkaline lipase by 50- fold. The effect of additives as well as the development of suitable culture conditions for lipase production is discussed.     

Large-Scale Chemical Similarity Networks for Target Profiling of Compounds Identified in Cell-Based Chemical Screens

Target identification is one of the most critical steps following cell-based phenotypic chemical screens aimed at identifying compounds with potential uses in cell biology and for developing novel disease therapies. Current in silico target identification methods, including chemical similarity database searches, are limited to single or sequential ligand analysis that have limited capabilities for accurate deconvolution of a large number of compounds with diverse chemical structures. Here, we present CSNAP (Chemical Similarity Network Analysis Pulldown), a new computational target identification method that utilizes chemical similarity networks for large-scale chemotype (consensus chemical pattern) recognition and drug target profiling. Our benchmark study showed that CSNAP can achieve an overall higher accuracy (>80%) of target prediction with respect to representative chemotypes in large (>200) compound sets, in comparison to the SEA approach (60–70%). Additionally, CSNAP is capable of integrating with biological knowledge-based databases (Uniprot, GO) and high-throughput biology platforms (proteomic, genetic, etc) for system-wise drug target validation. To demonstrate the utility of the CSNAP approach, we combined CSNAP''s target prediction with experimental ligand evaluation to identify the major mitotic targets of hit compounds from a cell-based chemical screen and we highlight novel compounds targeting microtubules, an important cancer therapeutic target. The CSNAP method is freely available and can be accessed from the CSNAP web server (http://services.mbi.ucla.edu/CSNAP/).     

Molecular detection of <Emphasis Type="Italic">Borrelia valaisiana</Emphasis>-related spirochetes from <Emphasis Type="Italic">Ixodes granulatus</Emphasis> ticks in Taiwan

Borrelia valaisiana-related spirochetes were detected for the first time in Ixodes granulatus ticks collected in Taiwan. The genetic identities of these detected spirochetes were determined by analyzing the gene sequences amplified by a genospecies-specific polymerase chain reaction assay based on the outer surface protein A (OspA) gene of B. burgdorferi sensu lato. Phylogenetic relationships were analyzed by comparing the sequences of OspA gene obtained from 35 strains of Borrelia spirochetes representing six genospecies of Borrelia. Eight major clades can be easily distinguished by neighbour-joining analysis and were congruent by maximum-parsimony method. Except one strain (KH-74), all these Borrelia spirochetes of Taiwan were genetically affiliated to the same clade with highly homogeneous sequences (97.8–100% similarity), and can be discriminated from other groups of B. valaisiana and other genospecies of Borrelia spirochetes with a sequence divergence ranging from 3 to 19.6%. Moreover, intraspecific analysis also revealed that three distinct groups are evident between the same species of B. valaisiana spirochetes detected in Taiwan. Our results provide the first evidence of B. valaisiana spirochetes detected in I. granulatus ticks collected in Taiwan and demonstrate that all these B. valaisiana spirochetes of Taiwan represent three major groups distinct from the European group of B. valaisiana spirochetes.     

Clinical Manifestations of Non-O1 Vibrio cholerae Infections

Background

Infections caused by non-O1 Vibrio cholera are uncommon. The aim of our study was to investigate the clinical and microbiological characteristics of patients with non-O1 V. cholera infections.

Methods

The clinical charts of all patients with non-O1 V. cholera infections and who were treated in two hospitals in Taiwan were retrospectively reviewed.

Results

From July 2009 to June 2014, a total of 83 patients with non-O1 V. cholera infections were identified based on the databank of the bacteriology laboratories of two hospitals. The overall mean age was 53.3 years, and men comprised 53 (63.9%) of the patients. Liver cirrhosis and diabetes mellitus were the two most common underlying diseases, followed by malignancy. The most common type of infection was acute gastroenteritis (n = 45, 54.2%), followed by biliary tract infection (n = 12, 14.5%) and primary bacteremia (n = 11, 13.3%). Other types of infection, such as peritonitis (n = 5, 6.0%), skin and soft tissue infection (SSTI) (n = 5, 6.0%), urinary tract infection (n = 3, 3.6%) and pneumonia (2, 2.4%), were rare. July and June were the most common months of occurrence of V. cholera infections. The overall in-hospital mortality of 83 patients with V. cholera infections was 7.2%, but it was significantly higher for patients with primary bacteremia, hemorrhage bullae, acute kidney injury, acute respiratory failure, or admission to an ICU. Furthermore, multivariate analysis showed that in-hospital mortality was significantly associated with acute respiratory failure (odds ratio, 60.47; 95% CI, 4.79-763.90, P = 0.002).

Conclusions

Non-O1 V. cholera infections can cause protean disease, especially in patients with risk factors and during warm-weather months. The overall mortality of 83 patients with non-O1 V. cholera infections was only 7.2%; however, this value varied among different types of infection.