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This study examined the effects of Maillard reaction products reacted by casein and lactose (cMRP) and of cMRP fermented by Lactobacillus fermentum H9 (FcMRP) on hypolipidemic and antiinflammatory effects in rats fed a high-fat and high-cholesterol diet (HD). The HD-fed rats had significantly increased hepatic triglyceride concentrations compared with the rats fed a normal diet. It was shown that treatment with simvastatin, L. fermentum H9 (H9), cMRP, and F-cMRP decreased total triglycerides in the liver compared with the HD group. On histological analysis, a reduction of lipid accumulation in the liver and aortic tissues was observed in the cMRP, F-cMRP, and H9-fed rats. Also, F-cMRP and cMRP reduced intima-media thickness in the HD group. In addition, the H9, cMRP, and F-cMRP treatments significantly reduced the expression levels of ICAM-1 and VCAM-1, but not of MCP-1. In particular, the expressions of ICAM-1 and VCAM-1 were significantly decreased in the F-cMRP group compared with the HD group. These results of the present study suggest that cMRP and F-cMRP in dairy foods could potentially be used to prevent or treat cardiovascular diseases, especially atherosclerosis. [ABSTRACT FROM AUTHOR]

Concurrently, several porcine epidemic diarrhea virus (PEDV) variants are circulating in US swine farms, including the original US and the spike insertion-deletion (S-INDEL) strains. In this study, reverse transcription (RT)-PCR assays for the detection and differentiation of different US PEDV variants were developed based on the differences in the S1 domain of the spike (S) gene. This assay successfully differentiated three PEDV strains: PC22A (the original US virulent), Iowa106 (S-INDEL), and PC177 (S-197DEL) that was derived from cell culture adaptation and has a 197 amino acid-deletion in the S1 domain. The assays did not amplify the porcine deltacoronavirus OH-FD22 strain or transmissible gastroenteritis virus Miller strain. It is the first report on the development of RT-PCR assays allowing the detection and differentiation of all major types of US PEDV variants. [ABSTRACT FROM AUTHOR]

Abstract: Bcl-xL is a pro-survival member of the Bcl-2 family that plays indispensable roles in regulating cell survival and apoptosis. It is overexpressed in many malignant tumors including colorectal cancer (CRC). However, it is still unclear if Bcl-xL can be used as an independent molecular marker for predicting the prognosis of CRC patients. In this study, reverse transcription-PCR assay was performed to detect the expression of Bcl-xL mRNA in CRC and corresponding non-tumor colon tissues. Immunohistochemistry was performed to detect the immunolocalization of Bcl-xL protein in sixty-eight primary CRC tissue samples. The association between Bcl-xL protein expression and clinicopathological factors of CRC patients was analyzed and the survival was assessed by the Kaplan–Meier method and proportional hazards model. The averaged level of Bcl-xL mRNA expression in CRC tissues (0.85±0.13) was significantly higher than that in non-tumor colon tissues (0.08±0.02). Immunohistochemical staining showed that the Bcl-xL protein was mainly located in the cytoplasm of tumor cells. The level of Bcl-xL protein expression was closely correlated with tumor differentiation (P =0.002), lymph node metastasis (P =0.010), venous permeation (P =0.004), and Duke''s classification (P =0.021). Furthermore, patients with high Bcl-xL expression showed poorer overall survival than those with low Bcl-xL expression (P =0.016). Univariate and multivariate analysis indicated that the status of Bcl-xL protein expression might be an independent prognostic marker for CRC patients (P =0.032). Taken together, immunohistochemical assessment of status of Bcl-xL protein may offer a valuable approach for predicting survival after curative surgery for colorectal cancer. [Copyright &y& Elsevier]

Abstract: Enteric viruses are important agents of foodborne disease. Unfortunately, robust, quantitative methods for sampling and analysis of enteric and other viruses in processed or complex foods are not well-established. As a result, epidemiologically determined etiologies or pathogen sources in foodborne outbreaks are rarely confirmed by virological analysis. In this study, an acid-adsorption elution concentration (AEC) method previously used to monitor virus occurrence and investigate enteric virus outbreaks in shellfish was adapted for examination of processed food items, namely tomato sauce and blended strawberries. Hepatitis A virus (HAV), poliovirus, and coliphage MS2 (MS2) were seeded in 10 or 30 g samples of tomato sauce or blended strawberries, recovered by AEC, and quantified by cell culture infectivity assay. In addition, nested reverse transcription-polymerase chain reaction (RT-PCR) and TaqMan RT-PCR assays were used to detect HAV RNA. Viruses were efficiently adsorbed to foods as an initial concentration step, with infectious HAV and MS2 adsorption of 67% and 93%, respectively, to tomato sauce, and 89% and 99%, respectively, to blended strawberries. Forty-three to 65% of HAV and poliovirus were subsequently eluted and recovered from tomato sauce using 0.5 M threonine, pH 7.2. The lower limits of HAV detection were at initial seeding levels of 14 PFU/g of tomato sauce and 33 PFU/g of blended strawberries. Unlike TaqMan RT-PCR, nested RT-PCR was not inhibited by undiluted final RNA extracts of tomato sauce or blended strawberries. The successful adaptation of the AEC method for enteric and other virus recovery, quantitation and detection in processed foods demonstrates its potential for use in the investigation of foodborne outbreaks of viral etiology and for validation of virus disinfection and sanitary processing procedures used by the food industry. [Copyright &y& Elsevier]

Abstract: The basic helix-loop-helix/Per-Arnt-Sim (bHLH-PAS) homology protein family is an important class of transcriptional regulators that are involved in a wide variety of biological processes. In a previous study, we characterized a novel bHLH-PAS factor ‘NXF’ as a new member of the mammalian bHLH-PAS family. If model animals have ortholog genes for NXF, they might have advantages for characterization of its function and regulation in vivo. Here we document cDNA cloning and characterization of putative NXF homolog genes (for NXF-like-factors) in Drosophila and C. elegans. Some conservation of the exon–intron gene architecture was noted in the bHLH-PAS domains among NXF-like-factors and human NXF. The NXF-like-factors selectively interacted with an Arnt ortholog in vitro assay. Moreover, they selectively associated with the mammalian NXF:Arnt2 binding element in the presence of the Arnt ortholog, and synergistically activated a mammalian NXF:Arnt-responsive promoter. These data strongly suggest that the Drosophila and C. elegans NXF-like-factors reported here are homologs of mammalian NXF. Further elucidation of whether they are true orthologs with the same function in vivo is now necessary. [Copyright &y& Elsevier]

Abstract: In our present study we used preparations from Xenopus laevis lungs to perform electrophysiological Ussing chamber measurements, unidirectional flux measurements, and employed molecular approaches to elucidate the presence and function of a cystic fibrosis transmembrane conductance regulator (CFTR) homolog in this tissue. Application of different CFTR blockers (NPPB (5-nitro-2-(3-phenylpropylamino)benzoic acid), niflumic acid (NFA), glibenclamide, lonidamine, CFTRinh-172) to the apical side of the tissues was able to significantly decrease the measured short circuit current (I SC) indicating a Cl− secretion due to luminal located CFTR channels. This was further supported by a net 36Cl− secretion determined by radioactive tracer flux experiments. Further, Xenopus pulmonary epithelia responded to apical chlorzoxazone exposure – a CFTR activator – and this activated current was inhibited by CFTRinh-172. We performed reverse transcription-PCR (RT-PCR) and Western blot analysis and with both approaches we found characteristic signals indicating the presence of a CFTR homolog in Xenopus lung. In addition, we were able to detect CFTR in apical membranes of Xenopus lung slices with immunohistological techniques. We conclude that Xenopus lung epithelium exhibits functional CFTR channels and that this tissue represents a valuable model for the investigation of ion transport properties in pulmonary epithelia. [Copyright &y& Elsevier]

Abstract: Background: Human respiratory coronavirus (hCoV) HKU1 infections were reported for the first time in 2005 in Hong Kong. Objective: To investigate epidemiological, clinical, and diagnostic features of HKU1 infections. Study design: Longitudinal, prospective study from November 2005 through May 2006 in a hospitalised patient population. Results: Overall, 48/426 (11.3%) patients were found to be infected by hCoV acute respiratory tract infections (ARTI). Of these, 10 (19.2%) were caused by HKU1 (6 single infections and 4 coinfections) during the period January–May 2006. Diagnosis was made by using RT-PCR for all four hCoVs, and in parallel, in-house developed group-specific monoclonal antibodies (MAbs) for HKU1 and 229E. HKU1-specific MAb was able to retrospectively identify 8 of 10 HKU1 strains detected by RT-PCR. Phylogenetic analysis showed that four HKU1 strains were genotype A and six genotype B. In HKU1-infected patients, the predominant clinical symptom was rhinorrhea (nine patients). Within group II hCoV, HKU1-infected patients had a significantly lower rate of lower ARTI compared to OC43-infected patients. Conclusion: HKU1 hCoV strains circulated in northern Italy during the winter–spring season 2005–2006. Both HKU1 genotypes were detected. HKU1-specific MAb may contribute to the rapid diagnosis of HKU1 infections currently performed by RT-PCR. [Copyright &y& Elsevier]

The detection of hepatitis A virus in shellfish by reverse transcription-nested polymerase chain reaction (RT-nested PCR) is hampered mainly by low levels of virus contamination and PCR inhibitors in shellfish. In this study, we focused on getting a rapid and sensitive processing procedure for the detection of HAV by RT-nested PCR in clam samples (Tapes spp.). Two previously developed processing methods for virus concentration in shellfish have been improved upon and compared. The first method involves acid adsorption, elution, polyethylene glycol (PEG) precipitation, chloroform extraction and PEG precipitation. The second method is based on elution with a glycine buffer at pH 10, chloroform extraction and concentration by ultracentrifugation. Final clam concentrates were processed by RNA extraction or immunomagnetic capture of viruses (IMC) before the RT-nested PCR reaction. Both methods of sample processing combined with the RNA extraction from the concentrates were very efficient when they were assayed in seeded and naturally contaminated samples. The results show that the first method was more effective in removal inhibitors and the second was simpler and faster. The IMC of HAV from clam concentrates processed by method 1 was revealed to be a very effective method of simultaneously removing residual PCR inhibitors and of concentrating the virus. [Copyright &y& Elsevier]