A physiologically based kinetic (PBK) model describing plasma concentrations of quercetin and its metabolites in rats

Biological activities of flavonoids in vivo are ultimately dependent on the systemic bioavailability of the aglycones as well as their metabolites. In the present study, a physiologically based kinetic (PBK) model was developed to predict plasma concentrations of the flavonoid quercetin and its metabolites and to tentatively identify the regiospecificity of the major circulating metabolites. The model was developed based on in vitro metabolic parameters and by fitting kinetic parameters to literature available in vivo data. Both exposure to quercetin aglycone and to quercetin-4′-O-glucoside, for which in vivo data were available, were simulated. The predicted plasma concentrations of different metabolites adequately matched literature reported plasma concentrations of these metabolites in rats exposed to 4′-O-glucoside. The bioavailability of aglycone was predicted to be very low ranging from 0.004%-0.1% at different oral doses of quercetin or quercetin-4′-O-glucoside. Glucuronidation was a crucial pathway that limited the bioavailability of the aglycone, with 95–99% of the dose being converted to monoglucuronides within 1.5–2.5 h at different dose levels ranging from 0.1 to 50 mg/kg bw quercetin or quercetin-4′-O-glucoside. The fast metabolic conversion to monoglucuronides allowed these metabolites to further conjugate to di- and tri-conjugates. The regiospecificity of major circulating metabolites was observed to be dose-dependent. As we still lack in vivo kinetic data for many flavonoids, the developed model has a great potential to be used as a platform to build PBK models for other flavonoids as well as to predict the kinetics of flavonoids in humans.     

Molecular characterization of VP4, VP6, VP7, NSP4, and NSP5/6 genes identifies an unusual G3P[10] human rotavirus strain

An unusual strain of human rotavirus G3P[10] (CMH079/05) was detected in a stool sample of a 2‐year‐old child admitted to the hospital with severe diarrhea in Chiang Mai, Thailand. Analysis of the VP7 gene sequence revealed highest identities with unusual human rotavirus G3 strain CMH222 at 98.7% on the nucleotide and 99.6% on the amino acid levels. Phylogenetic analysis of the VP7 sequence confirmed that the CMH079/05 strain formed a cluster with G3 rotavirus reference strains and showed the closest lineage with the CMH222 strain. Analysis of partial VP4 gene of CMH079/05 revealed highest degree of sequence identities with P[10] rotavirus prototype strain 69M at nucleotide and amino acid levels of 92.9% and 94.6%, respectively. Phylogenetic analysis of the VP4 sequence revealed that CMH079/05 and 69M clustered closely together in a monophyletic branch separated from other rotavirus genotypes. To our knowledge, this is a novel G–P combination of G3 and P[10] genotypes. In addition, analyses of VP6, NSP4, and NSP5/6 genes revealed these uncommon genetic characteristics: (i) the VP6 gene differed from the four other known subgroups; (ii) the NSP4 gene was identified as NSP4 genetic group C, an uncommon group in humans; and (iii) the NSP5/6 gene was most closely related with T152, a G12P[9] rotavirus previously isolated in Thailand. The finding of uncommon G3P[10] rotavirus in this pediatric patient provided additional evidence of the genetic diversity of human group A rotaviruses in Chiang Mai, Thailand. J. Med. Virol. 81:176–182, 2009. © 2008 Wiley‐Liss, Inc.     

Circulating of human bocavirus 1, 2, 3, and 4 in pediatric patients with acute gastroenteritis in Thailand

Human bocavirus (HBoV) is a newly identified human parvovirus that associated with respiratory and gastrointestinal diseases. Epidemiological surveillance of HBoV was conducted on fecal specimens collected from hospitalized children with diarrhea in Chiang Mai, Thailand in 2011. Among a total of 222 fecal specimens tested, 17 (7.7%) were positive for HBoV by PCR. Of the 17 HBoV positive samples, double- or triple-infections together with other enteric viruses were found in 10 (58.8%) pediatric patients, while monoinfection with HBoV alone was detected in seven (41.2%) cases. Mixed infection among HBoV with norovirus GII was frequently observed in this population. The partial VP1 nucleotide sequences of all 17 HBoV strains demonstrated that all four species of HBoV were found in the specimens tested. Eleven strains were HBoV1. Other three strains showed the sequence identity with HBoV2, which were most closely related to the HBoV2A. In addition, other two HBoV strains showed the highest level of nucleotide sequence identity with the HBoV3. It was surprisingly to observe that one Thai HBoV strain showed a unique characteristic similar to the HBoV4, a rare species of HBoV found in acute gastroenteritis patients. In summary, this study presents the genetic background information of HBoV circulated in acute gastroenteritis children in Chiang Mai, Thailand and it was clearly demonstrated that HBoVs circulated in this area were genetically diverse as all four species of HBoVs (HBoV1-4) were detected in the fecal specimens collected from pediatric patients admitted to the hospitals in this area.     

The unique characteristics of Thai Leber hereditary optic neuropathy: analysis of 30 G11778A pedigrees

Leber hereditary optic neuropathy (LHON) is characterized by acute or subacute bilateral visual loss, and affects mostly young males. The most common mitochondrial DNA mutation responsible for LHON worldwide is G11778A. Despite different genetic backgrounds, which are believed to influence the disease expression, most features of LHON are quite common in different populations. However, there seem to be a few ethnic-specific differences. Analyses of our 30 G11778A LHON pedigrees in Thailand showed some characteristics different from those of Caucasians and Japanese. In particular, our pedigrees showed a lower male to female ratio of affected persons (2.6:1) and much higher prevalence of G11778A blood heteroplasmy (37% of the pedigrees contained at least one heteroplasmic G11778A individual). Heteroplasmicity seemed to influence disease manifestation in our patients but did not appear to alter the onset of the disease. The estimated overall penetrance of our G11778A LHON population was 37% for males and 13% for females. When each of our large pedigrees were considered separately, disease penetration varied from 9 to 45% between the pedigrees, and also varied between different branches of the same large pedigree. Survival analysis showed that the secondary LHON mutations G3316A and C3497T had a synergistic deleterious effect with the G11778A mutation, accelerating the onset of the disease in our patients.     

Identification of a novel 74-kiloDalton immunodominant antigen of Pythium insidiosum recognized by sera from human patients with pythiosis

The oomycetous, fungus-like, aquatic organism Pythium insidiosum is the etiologic agent of pythiosis, a life-threatening infectious disease of humans and animals that has been increasingly reported from tropical, subtropical, and temperate countries. Human pythiosis is endemic in Thailand, and most patients present with arteritis, leading to limb amputation and/or death, or cornea ulcer, leading to enucleation. Diagnosis of pythiosis is time-consuming and difficult. Radical surgery is the main treatment for pythiosis because conventional antifungal drugs are ineffective. The aims of this study were to evaluate the use of Western blotting for diagnosis of human pythiosis, to identify specific immunodominant antigens of P. insidiosum, and to increase understanding of humoral immune responses against the pathogen. We performed Western blot analysis on 16 P. insidiosum isolates using 12 pythiosis serum samples. These specimens were derived from human patients with pythiosis who had different forms of infection and lived in different geographic areas throughout Thailand. We have identified a 74-kDa immunodominant antigen in all P. insidiosum isolates tested. The 74-kDa antigen was also recognized by sera from all patients with pythiosis but not by control sera from healthy individuals, patients with thalassemia, and patients with various infectious diseases, indicating that Western blot analysis could facilitate diagnosis of pythiosis. Therefore, the 74-kDa antigen is a potential target for developing rapid serodiagnostic tests as well as a therapeutic vaccine for pythiosis. These advances could lead to early diagnosis and effective treatment, crucial factors for better prognosis for patients with pythiosis.     

Microwave-assisted synthesis of Ag/ZnO nanoparticles using Averrhoa carambola fruit extract as the reducing agent and their application in cotton fabrics with antibacterial and UV-protection properties

This is the first time Averrhoa carambola fruit extract has been used as a reducing agent to synthesize Ag/ZnO composites for coating cotton to develop antibacterial activity and UV protection under domestic microwave irradiation. The effects of the molar concentration of silver nitrate solutions, applied power, reaction duration, and pH on the yield of nanoparticles were determined. The treated fabrics were subjected to the investigation of surface morphology and chemical structure using SEM and EDX techniques, respectively. The antibacterial activity of the ZnO NPs and the Ag/ZnO nanocomposite coated on cotton fabric was evaluated against E. coli and S. aureus using the agar well diffusion method. The results revealed good antibacterial activity in the cotton fabric treated with the Ag-doped ZnO composite. The stability of the Ag/ZnO nanocomposite coated fabrics was determined by a wash durability test, the results of which demonstrated that this fabric could retain good antibacterial activity even after 20 wash cycles. The UV-blocking capacity of the treated fabrics was evaluated based on the ultraviolet protection factor (UPF) value determined in the range of 280–400 nm. The UPF value determined for the Ag/ZnO-coated fabric was 69.67 ± 1.53, which indicated an excellent ability to block UV radiation. Collectively, these results demonstrated the Ag/ZnO nanocomposite prepared in the present study as a promising material for preparing textiles with good antibacterial activity and UV protection.

This is the first time Averrhoa carambola fruit extract has been used as a reducing agent to synthesize Ag/ZnO composites for coating cotton to develop antibacterial activity and UV protection under domestic microwave irradiation.