Retrovirus‐mediated gene transfer of the human beta‐globin gene into hematopoietic stem cells is an attractive approach to the therapy of human beta‐globin gene disorders. However, expression of the transduced beta‐globin gene linked to its proximal cis‐acting sequences (‐0.8 to +0.3 kb from the cap site) is considerably below the level required for a significant therapeutic effect. The discovery of the beta‐locus control region (beta‐LCR), organized in four major DNase I hypersensitive sites far upstream of the human beta‐like globin gene cluster, provided a potential means to achieve a high level of expression of a linked human beta‐globin gene, but initial attempts to incorporate beta‐LCR derivatives in retroviral vectors resulted in the production of low‐titer viruses with multiple rearrangements of the transmitted proviral structures. We now describe how extensive mutagenesis of the transduced beta‐globin gene, eliminating a 372 bp intronic segment and multiple reverse polyadenylation and splicing signals, increases viral titer significantly and restores stability of proviral transmission upon infection of cell lines and bone marrow‐repopulating cells. These optimized vectors have enabled us to analyze the expression properties of various retrovirally transduced beta‐LCR derivatives in dimethylsulfoxide‐induced murine erythroleukemia cells and to achieve ratios of human beta‐globin/murine beta maj‐globin mRNA, on a per gene basis, as high as 80%.

Human y-globin and -globin chains have been previously identified as strong inhibitors of the polymerization of hemoglobin S, in contrast to the ,8-globin chain, which exerts only a moderate antisickling effect. However, -y-globin and 8-globin are normally expressed at very low levels in adult erythroid cells, in contrast to j8-globin. We report the design of a j3-globin/8-globin hybrid gene, j3/6 sickle cell inhibitor 1 (13/6-SCI1) and its transduction by retrovirus-mediated gene transfer. The 8/6-SCII-encoding gene retains the overall structure of the human j8-globin gene, while incorporating specific amino acid residues from the 6 chain previously found responsible for its enhanced antisickling properties. To achieve high expression levels of ./ SCI1 in adult erythrocytes, the hybrid gene was placed under the transcriptional control of the human f3-globin promoter and the DNase I hypersensitive site 2 of the human 3 locus control region. High-titer retroviruses were generated, and stable proviral transmission was achieved in infected cells. The mRNA expression levels of the .8/6-SCI1 gene in infected, dimethyl sulfoxide-induced murine erythroleukemia cells approached 85% of the endogenous murine f3maj-globin mRNA, on a per gene basis, evidence that high gene expression levels were achieved in adult erythroid cells. Further evaluation of this strategy in transgenic animal models of sickle cell disease should assess its efficacy for the gene therapy of human patients.

Acute gastroenteritis (AGE), which is one of the most common diseases worldwide, primarily occurs in infants and young children in both developed and developing countries. To investigate the prevalence of AGE in Korea, 6,788 stool specimens collected from hospitalized patients with AGE in Seoul, Korea from March 2004 to June 2007 were analyzed by enzyme immunoassay, reverse transcription-PCR, DNA sequencing and phylogenetic analysis. Enteric viruses and bacteria were detected in 2,955 (43.5%) and 1,389 (20.5%) specimens, respectively. Among the enteric viruses detected, rotavirus (19.7%) and norovirus (18.9%) were the predominant causative agents, followed by adenovirus (2.5%) and astrovirus (2.4%). Staphylococcus aureus was the most commonly observed bacteria (8.0~19.2%). The epidemic peaks of the enteric viruses were October to December for norovirus, January to May for rotavirus, and August to October for adenovirus. The seasonal activity of rotavirus was shifted from winter to late spring. However, astrovirus did not display seasonal activity in this study. Although viral AGE primarily occurred in patients younger than 5 years of age, the incidence of viral AGE in children aged 6 to 14 years was significant. The results of this study will contribute to the currently available epidemiological data and improve public health and hygiene via amelioration of diagnostic methods and longitudinal surveillance.

In this study, inductively coupled plasma-mass spectrometry (ICP-MS) was used to determine the concentration of 15 elements (Mg, Al, K, Ca, Cr, Mn, Co, Ni, Cu, Zn, Rb, Sr, Cd, Ba, and Pb) of sesame seeds. Multivariate analysis was then performed to discriminate the origin of sesame seeds. Korean (48), Chinese (44), and Indian (21) samples were used to develop the calibration model. Another 10 samples were used to validate this model. All elements were significantly different (<0.05) among the samples from three countries, and all elements were subjected to both principal component analysis (PCA) and discriminant analysis. The concentrations of multi-element showed a trend of clustering according to the origin of samples based on PCA. They showed a discrimination rate of 92.0% in the discriminant analysis. The results demonstrated that a combination of ICP-MS multi-element determination and multivariate analysis could be used to discriminate the sesame seed origin.