DETECTION AND ENUMERATION OF H2S+ BACTERIA: APPLICATION TO SHEWANELLA PUTREFACIENS (CIP)

H₂S⁺ bacteria responsible for the degradation of sulfur-containing amino acids of fish muscle are currently little used to evaluate the microbiological pal quality of fish. Shewanella putrefaciens greatly predominates in this flora, and was therefore used to define a suitable culture method and medium. Inoculations by the Spiral surface method at 25C, with an incubation of 72h, gave the best counts on a medium containing two sources of sulfur (organic and inorganic) for H₂S⁺ bacteria. The culture medium and the NaCl concentration were determinant in the evaluation of this flora. At present there is no standard medium which meets these requirements.     

Adenovirus serotype determines association and localization of the large E1B tumor antigen with cellular tumor antigen p53 in transformed cells.

The distribution and stability of the cellular tumor antigen p53 were studied in baby rat kidney cells transformed by region E1 sequences of nononcogenic adenovirus (Ad) type 5 (Ad5) or oncogenic type 12 (Ad12). In transformed cells expressing the large E1B T antigen of Ad5, p53 was associated with this T antigen. The complexed proteins were concentrated in a cytoplasmic body, which has been shown to consist of a cluster of 8-nm filaments (A. Zantema et al., Virology 142:44-58, 1985). In transformed cells expressing the E1B region of Ad12, however, no association between the viral large T antigen and p53 was detectable. In the latter case, both proteins were found almost exclusively in the nucleus. The stability of p53 in both Ad5- and Ad12-transformed cells was increased relative to that in primary cells or cells immortalized by the E1A region only. Thus, the increased stability of p53 in Ad-transformed cells is not caused by association with a viral T antigen, but it correlates with expression of E1B and with morphological transformation.     

STING Millennium Suite: integrated software for extensive analyses of 3d structures of proteins and their complexes

Background  

The integration of many aspects of protein/DNA structure analysis is an important requirement for software products in general area of structural bioinformatics. In fact, there are too few software packages on the internet which can be described as successful in this respect. We might say that what is still missing is publicly available, web based software for interactive analysis of the sequence/structure/function of proteins and their complexes with DNA and ligands. Some of existing software packages do have certain level of integration and do offer analysis of several structure related parameters, however not to the extent generally demanded by a user.     

Insect fragments in flour: relationship to lesser grain borer (Coleoptera: Bostrichidae) infestation level in wheat and rapid detection using near-infrared spectroscopy

We determined that the number of insect fragments, quantified using the standard flotation method, in flour milled from wheat infested with larvae, pupae, or preemergent adults of the lesser grain borer, Rhyzopertha dominica (F.), was proportional to infestation level. Wheat infested with a single preemergent adult contributed 28 and 10x as many fragments as wheat infested with a single larva or pupa, respectively. Using regression models that were developed from these data, we predicted that the maximum infestation level that would result in flour with fragment counts below the Food and Drug Administration defect action level (75 fragments/50 g of flour) was 0.95 and 1.5% (380-640 infested kernels/kg of wheat) for pupae and larvae, but it decreased to 0.05% (20 infested kernels/kg) when the grain was infested with preemergent adults. We also reexamined the accuracy and sensitivity of near-infrared spectroscopy (NIRS) for detecting insect fragments in flour by testing three different NIR spectrometers. NIRS-predicted numbers of insect fragments were correlated with the actual number of fragments. NIRS is less precise than the standard flotation method, but it is rapid, nondestructive, does not require extensive sample preparation, and could easily be automated for a more sophisticated sampling protocol for flour based on prescreening samples with NIRS followed up by use of the standard flotation method when necessary.     

Pathogenicity of Epicoccum sorghinum towards dragon fruits (Hylocereus species) and in vitro evaluation of chemicals with antifungal activity

Recent reports of plant diseases that result in yield reduction and increasing demand for dragon fruits raise concerns of fruit supply shortage. Emerging plant diseases may play an important role in increasing yield losses and reducing the availability of stem cuttings (source of planting materials). Understanding the aetiology of current and new diseases of dragon fruit is important to address production issues and to formulate effective disease control measures. This study reports Epicoccum sorghinum as a potential emerging pathogen of dragon fruit. Epicoccum sorghinum MBDF0024a was isolated from dragon fruit stems (Hylocereus monacanthus) showing brown spot symptoms. DNA sequence of the internal transcribed spacer (ITS-rDNA), beta tubulin and actin gene regions of fungal isolate MBDF0024a had high similarities to E. sorghinum stains. Epicoccum sorghinum MBDF0024a was pathogenic to three cultivated dragon fruit species (Hylocereus undatus, H. monacanthus and H. megalanthus) in repeated laboratory and glasshouse trials. Large brown lesions developed on 3-week-old inoculated rooted stem cuttings 3 days postinoculation (dpi). Yellowing of the lesion (advance part) started at five dpi, and at seven dpi, yellowing was observed in the stem. As there are no reported control measures for diseases caused by E. sorghinum, this study screened chemicals with antifungal properties. A biopesticide containing B. subtilis (2 ml/400 ml), and chemicals isoprothiolane (2.25 ml/400 ml), mancozeb (2 g/400 ml) and pyraclostrobin (1 ml/400 ml) (chemical control) completely inhibited the in vitro growth of E. sorghinum MBDF0024a. The results establish E. sorghinum as a new and emerging pathogen of dragon fruit that could be a major yield-limiting disease if left uncontrolled. The biopesticide can be considered a fairly safe option for disease management, but glasshouse and field studies are needed for validation.     

The Proteolytic Potential of Normal Human Melanocytes: Comparison With Other Skin Cells and Melanoma Cell Lines

To understand the contribution of epidermal melanocytes in the proteolytic potential of human skin, we have studied melanocytes grown in a low-serum medium deprived of phorbol esters, cholera toxin, and other non-physiological supplements. We focused on the plasminogen activation system and certain matrix metalloproteinases (gelatinases). Supposing that the proteolytic activity of cells can influence binding to collagen matrix and its reorganization, we have analyzed these parameters as well. We found that human melanocytes secreted tissue-type plasminogen activator and utilised it to generate cell-bound plasmin. No urokinase-type plasminogen activator was detected in the cultures but its receptor was found in cell extracts. Both the 72 kDa and 92 kDa gelatinases were secreted by the cells and in equal amounts. In addition, melanocytes secreted the wide-spectrum proteinase inhibitor alpha-2-macroglobulin. Melanocytes cast into collagen matrices retained a rounded morphology, did not extend processes, and were unable to contract collagen lattices. As a control, these parameters were investigated in parallel in cultures of human keratinocytes, dermal fibroblasts, and two melanoma cell lines. The obtained characteristics suggest that normal human melanocytes are proteolytically active cells. This function may pertain to skin physiology and pathophysiology.     

Genome-wide Introgression Lines and their Use in Genetic and Molecular Dissection of Complex Phenotypes in Rice (Oryza sativa L.)

Tremendous efforts have been taken worldwide to develop genome-wide genetic stocks for rice functional genomic (FG) research since the rice genome was completely sequenced. To facilitate FG research of complex polygenic phenotypes in rice, we report the development of over 20 000 introgression lines (ILs) in three elite rice genetic backgrounds for a wide range of complex traits, including resistances/tolerances to many biotic and abiotic stresses, morpho-agronomic traits, physiological traits, etc., by selective introgression. ILs within each genetic background are phenotypically similar to their recurrent parent but each carries one or a few traits introgressed from a known donor. Together, these ILs contain a significant portion of loci affecting the selected complex phenotypes at which allelic diversity exists in the primary gene pool of rice. A forward genetics strategy was proposed and demonstrated with examples on how to use these ILs for large-scale FG research. Complementary to the genome-wide insertional mutants, these ILs opens a new way for highly efficient discovery, candidate gene identification and cloning of important QTLs for specific phenotypes based on convergent evidence from QTL position, expression profiling, functional and molecular diversity analyses of candidate genes, highlights the importance of genetic networks underlying complex phenotypes in rice that may ultimately lead to more complete understanding of the genetic and molecular bases of quantitative trait variation in rice. Supplementary material to this paper is available in electronic form at http://dx.doi.org/10.1007/s11103-005-8519-3     

Differentiating tobacco budworm and corn earworm using near-infrared spectroscopy

Near-infrared spectroscopy (NIRS) was used to develop a simple and quick technique to differentiate two economically important species, the tobacco budworm, Heliothis cirescens (F.), and corn earworm, Helicoverpa zea (Boddie), which are major pests of cotton, Gossypium hirsutum L., in the southern United States. In practice, it is difficult to distinguish the two species during their immature stages using morphological characteristics unless expensive microscopy equipment or trained technicians are available. The current studies demonstrated that the two species could be quickly and readily differentiated during early developmental stages, including egg and young larval (younger than third instar) stages, by using NIRS technology with up to 95% accuracy. NIRS technology could significantly improve pest diagnosis in cotton pest management.