Isolation and characterisation of selected lactic acid bacteria for improved processing of Nem chua, a traditional fermented meat from Vietnam.

Nem chua is a traditional lactic acid fermented meat of Vietnam that is consumed raw. During fermentation, the number of acid-producing bacteria increased significantly and by the final day (4th), it had reached approximately 8.4 log cfu/g, contributing to 94% of the total microbiota of the product. Out of 85 isolates obtained from the product, only 44 were confirmed Gram-positive and catalase-negative. These putative LAB isolates were purified and screened for antimicrobial and proteolytic activity. None of the isolates showed bacteriocin activity using the 'spot-on lawn' method, but showed antimicrobial activity against Lactobacillus sakei and Enterococcus faecium using deferred assay. Four isolates showed good proteolytic activities at pH 7.0. These isolates were identified as Lactobacillus plantarum using an API 50 CHL i.d. kit. Sequencing of the partial 16S rRNA gene amplified by primers 1101F (5'-AACGAGCGCAACCC-3') and 1407R (5'-GACGGGCGGTGTGTAC-3') showed 98% homology to Lactobacillus plantarum WCFS1.

Sympathetic control of cardiac myosin heavy chain gene expression.

Several neuroendocrine factors have been shown to influence the muscle phenotype. Various physiological reports have suggested the role of adrenergic nervous system for cardiac myosin heavy chain (MHC) expression. We have used cultured fetal rat heart myocytes to investigate the role of cAMP on the alpha- and beta-MHC gene expression. In low density cultures, addition of 1 mM 8 Br cAMP resulted in up regulation of alpha-MHC and down regulation of beta-MHC mRNA. This antithetic effect of cAMP depends on the basal expression of both expression of both MHC transcripts. In transient transfection analysis employing a series of alpha-MHC gene promoter/reporter constructs, we identified a 13 bp E-box M-CAT hybrid motif (EM element) which conferred a basal muscle specific and cAMP-inducible expression of the alpha-MHC gene. Data obtained from the mobility gel-shift analysis indicated that one of the factor(s) binding to the EM element is related to troponin T M-CAT binding factor (TEF-1). To test whether the protein binding to this sequence could be a substrate for cAMP-dependent phosphorylation, the cardiac nuclear proteins were preincubated in a kinase reaction buffer either with a catalytic subunit of PKA (CatPKA) or with cAMP, and binding activity of proteins to the EM element was evaluated by mobility gel shift assay. In a concentration dependent manner, a twofold increase in the intensity of the retarded band was observed. Furthermore, at 100 units of CatPKA, an additional band of faster mobility was observed which was not present either when phosphorylated nuclear extract was incubated with alkaline phosphatase or when ATP was absent in kinase reaction buffer. These results strongly suggest that factor(s) binding to the EM element is a substrate for cAMP dependent phosphorylation.

Fusarium mycotoxins (fumonisins, nivalenol, and zearalenone) and aflatoxins in corn from Southeast Asia.

Corn samples collected from the Philippines, Thailand, and Indonesia were surveyed for the natural occurrence of Fusarium mycotoxins (fumonisins, trichothecenes, and zearalenone) and aflatoxins. Fumonisins B1 and B2 were found in over 50% of corn samples in individual countries, and their co-occurrences with aflatoxins at the incidence of 48% were noted. In addition to these mycotoxins, a trichothecene, nivalenol, and an estrogen, zearalenone, both mycotoxins of Fusarium species, were detected in these Southeast Asian samples. This is the first report on the simultaneous occurrence of two carcinogenic mycotoxins, fumonisins and aflatoxins, together with Fusarium mycotoxins (nivalenol and zearlenone) in corn from Asian tropics.

Differential expression of ventricular-like myosin heavy chain mRNA in developing and regenerating avian skeletal muscles.

Based on previous immunological data, cross-reactivity of myosin heavy chain (MHC) with the ventricular (V) isoform was observed in primordia of avian skeletal muscles and in regenerating adult anterior latissimus dorsi (ALD) muscle. To determine whether this primordial (P) MHC is identical to adult V-MHC gene product, we have cloned and characterized the 3' portion of MHC cDNA that is expressed in ALD muscle at 3 d of regeneration. Comparison of nucleotide sequences between adult V-MHC and P-MHC cDNAs revealed more than 98% homology in the 3'-untranslated (UT) portions of these genes. The expression pattern of P-MHC was analyzed in adult regenerating muscles using total RNA from two fast muscles, posterior latissimus dorsi (PLD) and pectoralis major (PM), as well as from slow ALD and mixed fast/slow gastrocnemius muscles at 0, 1, 3, 4, 6, 9, and 14 d after cold injury. Identical results were obtained by RNase protection assays using either a probe specifying the coding region of adult V-MHC or a P-MHC probe encoding the carboxy end plus the 3'-UT region. The expected protected fragments were detected early from day 2 up to day 6 in ALD muscle. Similar rate of appearance, reaching the highest level at day 3, was observed in PLD, PM, and gastrocnemius muscles. However, the amount and the kinetics of disappearance differed among the various muscles analyzed. In contrast, during development, steady-state levels and kinetics of V-MHC mRNA expression were found to be alike in axial and appendicular muscles. These data strongly suggest the identity of P-MHC as the ventricular isoform and support the concept that expression of P-MHC mRNA is a common feature of developing as well as of all regenerating adult skeletal muscles. Interestingly, no expression of cardiac specific myosin light chain (MLC) 2A was observed after cold injury, suggesting independent regulatory pathways for the two kinds of myosin subunits.

Both muscle-specific and ubiquitous nuclear factors are required for muscle-specific expression of the myosin heavy-chain beta gene in cultured cells.

Expression of the myosin heavy-chain beta gene is controlled by multiple cis-acting regulatory elements in the 5' flanking region; two of these, referred to as A (-276 to -263) and B (-207 to -180), are essential for conferring muscle-specific activation on homologous and heterologous promoters. Here we report on the identification of nuclear protein factors that specifically bind to these two elements. By using the A element as a probe, as well as nuclear extracts from muscle cells, we found two protein-DNA complexes that displayed distinct bands in a gel mobility shift assay but had identical methylation interference patterns. One complex was present mainly in nuclear extracts from undifferentiated muscle and nonmuscle cells, whereas the other was observed mainly in nuclear extracts from differentiated muscle cells. Thus, the muscle-specific complex formation with the A element appears to be involved in determining tissue-specific expression. Furthermore, competition analysis demonstrated that the A-element-binding factors also bind to the muscle-CAT motif in the cardiac troponin T gene. By using the B element as a probe, we saw similar patterns of gel-shifted bands and methylation interference in nonmuscle and muscle nuclear extracts. In addition, both elements A and B were found to be necessary for tissue-specific expression, suggesting that the muscle-specific activation of the myosin heavy-chain beta gene may require interaction between a muscle-specific and a ubiquitous protein-DNA complex.

Developmental outcome of infants with severe intracranial-intraventricular hemorrhage and hydrocephalus with and without ventriculoperitoneal shunt.

Thirty-six infants who developed grade III and IV intraventricular hemorrhages during the neonatal period were followed up to determine their developmental quotient. All of these infants had ventriculomegaly and 15 of them required a ventriculoperitoneal (VP) shunt during the neonatal period prior to discharge from Intensive Care Nursery. The mean developmental quotient for the infants with the VP shunt was 67.93. The mean developmental quotient for the infants with ventriculomegaly but no VP shunt was 88.71 (P less than 0.02). Among the nonshunted group of infants, 13 (61.9%) had developmental quotients greater than 85, and among the shunted group 5 infants (33.3%) had developmental quotients greater than 85. Fifty percent of the total group of infants had normal developmental quotients at a mean chronological age of 16.25 +/- 7.5 months (and corrected age 14 months). Infants developing posthemorrhagic hydrocephalus and requiring VP shunts had a poorer developmental outcome compared to those who did not require shunts.