Methylated N-(4-N,N-dimethylaminobenzyl) chitosan for novel effective gene carriers

The objective of this study was to investigate the transfection efficiency of quaternized N-(4-N,N-dimethylaminobenzyl) chitosan, TM(47)-Bz(42)-CS, using the plasmid DNA encoding green fluorescent protein (pEGFP-C2) on human hepatoma cell lines (Huh7 cells), in comparison to quaternized chitosan (TM(43)-CS) and chitosan (CS). Factors affecting the transfection efficiency, such as the carrier/DNA weight ratio, the pH of the culture medium, and the presence of serum, have been investigated. The results revealed that TM(47)-Bz(42)-CS was able to condense with pDNA. As illustrated by the agarose gel electrophoresis, the complete complexes of TM(47)-Bz(42)-CS/DNA were formed at a weight ratio of above 0.5, whereas those of TM(43)-CS/DNA and CS/DNA were formed at a ratio of above 1. TM(47)-Bz(42)-CS showed superior transfection efficiency to TM(43)-CS and CS at all weight ratios tested. Higher transfection efficiency and gene expression were observed when the carrier/DNA weight ratios increased. The highest transfection efficiency was found at a weight ratio of 8. The results indicated that the improved gene transfection was due to the hydrophobic group (N,N-dimethylaminobenzyl) substitution on CS, which promoted the interaction and condensation with DNA, as well as N-quaternization, which increased the CS water solubility. During cytotoxicity studies, it was found that high concentrations of TM(47)-Bz(42)-CS and TM(43)-CS could decrease the Huh7 cell viability. In conclusion, this novel CS derivative, TM(47)-Bz(42)-CS, shows promising potential as a gene carrier by efficient DNA condensation and a mediated higher level of gene transfection in Huh7 cells.     

Incorporation of camptothecin into N-phthaloyl chitosan-g-mPEG self-assembly micellar system.

The capability of N-phthaloylchitosan-grafted poly (ethylene glycol) methyl ether (mPEG)(PLC-g-mPEG) to enhance the aqueous solubility and stability of the lactone form of camptothecin (CPT) was investigated. PLC-g-mPEG formed a core-shell micellar structure after dialysis of the polymer solutions in dimethyl sulfoxide (DMSO) or dimethylformamide (DMF) against water, with a critical micelle concentration (CMC) of 28 microg/ml. CPT was loaded into the inner core of the micelles by dialysis method. The results showed an increase in the CPT-loading amount with an increasing concentration of CPT. The stability of drug-loaded micelles was studied by gel-permeation chromatography (GPC), and their in vitro release behaviors were analyzed. Release of CPT from the micelles was sustained. When compared to the unprotected CPT, CPT-loaded PLC-g-mPEG micelles were able to prevent the hydrolysis of the lactone group of the drug. The kinetics of the CPT hydrolysis in human serum albumin (HSA) and fetal bovine serum (FBS) were pseudo-first order. The hydrolysis rate constants for CPT and CPT-loaded PLC-g-mPEG micelles in phosphate-buffered saline (PBS) pH 7.4, were 7.4 x 10(-3) min(-1) and 9.1 x 10(-3) h(-1), parallel to an increase in half-life of CPT from 94 min to 76.15 h, respectively.     

Comparison of colorimetry and electrothermal atomic absorption spectroscopy for the quantification of non-transferrin bound iron in human sera

This paper describes a comparison of two analytical techniques, one employing bathophenanthrolinedisulfonate (BPT), a most commonly-used reagent for Fe (II) determination, as chromogen and an electrothermal atomic absorption spectroscopy (ETAAS) for the quantification of non-transferrin bound iron (NTBI) in sera from thalassemic patients. Nitrilotriacetic acid (NTA) was employed as the ligand for binding iron from low molecular weight iron complexes present in the serum but without removing iron from the transferrin protein. After ultrafiltration the Fe (III)-NTA complex was then quantified by both methods. Kinetic study of the rate of the Fe (II)-BPT complex formation for various excess amounts of NTA ligand was also carried out. The kinetic data show that a minimum time duration (> 60 minutes) is necessary for complete complex formation when large excess of NTA is used. Calibration curves given by colorimetric and ETAAS methods were linear over the range of 0.15-20 microM iron (III). The colorimetric and ETAAS methods exhibited detection limit (3sigma) of 0.13 and 0.14 microM, respectively. The NTBI concentrations from 55 thalassemic serum samples measured employing BPT as chromogen were statistically compared with the results determined by ETAAS. No significant disagreement at 95% confidence level was observed. It is, therefore, possible to select any one of these two techniques for determination of NTBI in serum samples of thalassemic patients. However, the colorimetric procedure requires a longer analysis time because of a slow rate of exchange of NTA ligand with BPT, leading to the slow rate of formation of the colored complex.     

Occupational health and safety situation and research priority in Thailand

The aims of this article are to review the situation of occupational health and safety and to describe research priority in this field in Thailand. Thailand is one of newly industrialized countries in Southeast Asia. Approximately half of the working population is still in agriculture. The data from Thai Workmen's Compensation Fund showed that incidence rate of occupational injuries and diseases was 3-4% each year. Almost were occupational injuries from various accidents in workplaces. At least 3 relevant governmental agencies, including Ministry of Labour, Ministry of Public Health, and Ministry of Industry, are responsible in occupational health and safety in the country. Nowadays, those agencies collaborate and develop projects and activities to prevent and control of the problems. Because of lack of staff and other resources, research priority is needed and has been developed recently. The framework of research needed focuses on research and development such as how to improve occupational health and safety management at all levels, setting up and development of standard guidelines for health and environmental assessment, and implementation of suitable control measures in workplaces. Finally, improvement of research system in the country is essential to cope with new occupational health problems in the near future.     

Persistent post-occupational dermatitis

Wall and Gebauer (Contact Dermatitis 1991: 24: 241-243) first described persistent post-occupational dermatitis (PPOD) as ongoing dermatitis for which there is no obvious present cause, precipitated by prior occupational contact dermatitis (OCD). We propose that individuals exhibiting PPOD lose the capacity for resolution of their condition upon removal from exposure to causative agents and subsequently develop persistent dermatitis, which can be continual or intermittent. Accordingly, we suggest modification of criterion 6 of the OCD criteria developed by Mathias (J Am Acad Dermatol 1989: 20: 842-848): 'Removal from exposure initially leads to improvement of dermatitis, however, over time there may be incomplete or no improvement, despite removal from exposures at work'. To satisfy the definition of PPOD, individuals must meet at least 4 of the 7 criteria, including the altered criterion 6. We present 6 cases of PPOD exemplifying these scenarios, which met the altered Mathias criteria. In some cases, subsequent failure to recognize the initial work relatedness of their skin conditions resulted in the termination of workers' compensation benefits. This situation is particularly relevant in the Australian context. The diagnosis of PPOD needs to be considered in all individuals with work-initiated dermatitis who present with ongoing endogenous-like eczema.     

Polymer design and incorporation methods for polymeric micelle carrier system containing water-insoluble anti-cancer agent camptothecin

A water-insoluble anti-cancer agent, camptothecin (CPT) was incorporated to a polymeric micelle carrier system forming from poly(ethylene glycol)-poly(aspartate) block copolymers. Incorporation efficiency and stability were analyzed in correlation with chemical structures of the inner core-forming hydrophobic blocks as well as with incorporation methods. Among three incorporation methods (dialysis, emulsion and evaporation methods), an evaporation method brought about much higher CPT yields with less aggregation than the other two methods. By the evaporation method, CPT was incorporated to polymeric micelles in considerably high yields and with high stability using block copolymers possessing high contents of benzyl and methylnaphtyl ester groups as hydrophobic moieties. This indicates importance of molecular design of the hydrophobic block chain to obtain targeting using polymeric micelle carriers as well as importance of the drug incorporation method.     

Reactogenicity and immunogenicity of reduced antigen content diphtheria-tetanus-acellular pertussis vaccine (dTpa) administered as a booster to 4-6 year-old children primed with four doses of whole-cell pertussis vaccine

A trial to compare the reactogenicity and immunogenicity of a reduced antigen content diphtheria-tetanus-acellular pertussis (dTpa) vaccine with diphtheria-tetanus-whole-cell pertussis (DTPw) vaccine was conducted in Thailand. Three hundred and thirty children aged 4-6 years, primed with four doses of DTPw, received a single injection of either dTpa or DTPw. There was a significantly lower incidence of local and general reactions following dTpa than DTPw (P<0.001). One month after vaccination, 99.4 and 100% of all subjects had protective anti-diphtheria and -tetanus titers, respectively. The vaccine response rate to pertussis antigens was similar in both groups, with 96.9% versus 92.5% for anti-pertussis toxin (PT), 96.9% versus 97.5% for anti-filamentous hemagglutinin (FHA) and 95.1% versus 90.8% for anti-pertactin (PRN) in the dTpa and DTPw groups, respectively. For anti-BPT, the vaccine response in the dTpa group was 29.6% versus 94.4% for DTPw. In conclusion, the dTpa vaccine was as immunogenic and significantly better tolerated than DTPw. The new dTpa vaccine could improve coverage for routine booster vaccination in children and provide a good replacement for DTP vaccines at 4-6 years of age.     

Enhanced antitumor effect of camptothecin loaded in long-circulating polymeric micelles.

A water-insoluble antitumor agent, camptothecin (CPT) was successfully incorporated into polymeric micelles formed from poly(ethylene glycol)-poly(benzyl aspartate) block copolymers (CPT-loaded polymeric micelles). Antitumor effects and biodistribution of CPT-loaded micelles were evaluated in mice subcutaneously transplanted by colon 26 tumor cells. Tumor growth was significantly inhibited after a single i.v. injection of CPT-loaded polymeric micelles at doses of either 15 or 30 mg/kg. Efficacy of a single high-dose injection was comparable to low dose multiple injections. CPT loaded in polymeric micelles showed prolonged blood circulation and higher accumulation in tumors compared with CPT in solution. Polymeric micelle systems offer a stable and effective platform for cancer chemotherapy with CPT.