<Emphasis Type="Italic">α</Emphasis>-Amylase inhibitor-1 gene from <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> expressed in <Emphasis Type="Italic">Coffea arabica</Emphasis>plants inhibits α-amylases from the coffee berry borer pest

Background  

Coffee is an important crop and is crucial to the economy of many developing countries, generating around US70 billion per year. There are 115 species in the < i > Coffea < /i > genus, but only two, < i > C. arabica < /i > and < i > C. canephora < /i > , are commercially cultivated. Coffee plants are attacked by many pathogens and insect-pests, which affect not only the production of coffee but also its grain quality, reducing the commercial value of the product. The main insect-pest, the coffee berry borer ( < i > Hypotheneumus hampei < /i > ), is responsible for worldwide annual losses of around US70 billion per year. There are 115 species in the Coffea genus, but only two, C. arabica and C. canephora, are commercially cultivated. Coffee plants are attacked by many pathogens and insect-pests, which affect not only the production of coffee but also its grain quality, reducing the commercial value of the product. The main insect-pest, the coffee berry borer (Hypotheneumus hampei), is responsible for worldwide annual losses of around US500 million. The coffee berry borer exclusively damages the coffee berries, and it is mainly controlled by organochlorine insecticides that are both toxic and carcinogenic. Unfortunately, natural resistance in the genus Coffea to H. hampei has not been documented. To overcome these problems, biotechnological strategies can be used to introduce an α-amylase inhibitor gene (α-AI1), which confers resistance against the coffee berry borer insect-pest, into C. arabica plants.     

Synonymous codon substitutions affect ribosome traffic and protein folding during in vitro translation

To investigate the possible influence of the local rates of translation on protein folding, 16 consecutive rare (in Escherichia coli) codons in the chloramphenicol acetyltransferase (CAT) gene have been replaced by frequent ones. Site-directed silent mutagenesis reduced the pauses in translation of CAT in E. coli S30 extract cell-free system and led to the acceleration of the overall rate of CAT protein synthesis. At the same time, the silently mutated protein (with unaltered protein sequence) synthesized in the E. coli S30 extract system was shown to possess 20% lower specific activity. The data suggest that kinetics of protein translation can affect the in vivo protein-folding pathway, leading to increased levels of protein misfolding.     

Structural characterization of Saccharomyces cerevisiae prion-like protein Ure2

Sacchromyces cerevisiae prion-like protein Ure2 was expressed in Escherichia coli and was purified to homogeneity. We show here that Ure2p is a soluble protein that can assemble into fibers that are similar to the fibers observed in the case of PrP in its scrapie prion filaments form or that form on Sup35 self-assembly. Ure2p self-assembly is a cooperative process where one can distinguish a lag phase followed by an elongation phase preceding a plateau. A combination of size exclusion chromatography, sedimentation velocity, and electron microscopy demonstrates that the soluble form of Ure2p consists at least of three forms of the protein as follows: a monomeric, dimeric, and tetrameric form whose abundance is concentration-dependent. By the use of limited proteolysis, intrinsic fluorescence, and circular dichroism measurements, we bring strong evidence for the existence of at least two structural domains in Ure2p molecules. Indeed, Ure2p NH2-terminal region is found poorly structured, whereas its COOH-terminal domain appears to be compactly folded. Finally, we show that only slight conformational changes accompany Ure2p assembly into insoluble high molecular weight oligomers. These changes essentially affect the COOH-terminal part of the molecule. The properties of Ure2p are compared in the discussion to that of other prion-like proteins such as Sup35 and mammalian prion protein PrP.     

Serologic evidence of West Nile virus infection in black bears (Ursus americanus) from New Jersey

Serum samples obtained from 51 free-ranging black bears (Ursus americanus) in northwestern New Jersey in February and March 2002 were analyzed for neutralizing antibodies to West Nile virus (WNV) and St. Louis encephalitis virus. Three (6%) of the black bears tested positive for WNV-neutralizing antibodies. One additional sample was positive for flavivirus-neutralizing antibodies but could not be differentiated for a specific virus type. This is the first report of WNV infection in black bears.     

The zipper model of translational control: a small upstream ORF is the switch that controls structural remodeling of an mRNA leader

Transport of the essential amino acids arginine and lysine is critical for the survival of mammalian cells. The adaptive response to nutritional stress involves increased translation of the arginine/lysine transporter (cat-1) mRNA via an internal ribosome entry site (IRES) within the mRNA leader. Induction of cat-1 IRES activity requires both translation of a small upstream open reading frame (uORF) within the IRES and phosphorylation of the translation initiation factor eIF2alpha. We show here that translation of the upstream ORF unfolds an inhibitory structure in the mRNA leader, eliciting a conformational change that yields an active IRES. The IRES, whose activity is induced by amino acid starvation, is created by RNA-RNA interactions between the 5' end of the leader and downstream sequences. This study suggests that the structure of the IRES is dynamic and regulation of this RNA structure is a mechanism of translational control.     

Screening of fixed prosthodontic dentures after five years of use in relation to material and construction

The aim of this study was to screen patients with fixed prosthodontic appliances that were in oral cavity for a period of 5 years or more and to assess clinically and radiologically root caries, gingival recession, pocket formation, alveolar ridge resorption, as well as gingival (GI) and plaque index (PI). The aim also was to find out the differences between materials and constructions, between abutment and non-abutment teeth, and to find out the need for replacement. A total of 260 patients and their orthopantomograms were examined, with a total of 2,265 teeth, 610 being bridge abutments and 246 being crowns. The most frequent were metal+ acrylic veneer crowns or bridges. Root caries was found under the abutments in 10-20%; however abutments with ceramic crowns had the lowest percentage of caries (p < 0.01). Alveolar ridge resorption, pocket formation deeper than 3 mm and gingival recession of various degree was found in 50% of the cases, again with the lowest percentage of ceramic-fused-to-metal appliances (p < 0.01). Pocket depth was registered in significantly higher percentage in metal-acrylic veneer appliances compared to natural teeth (p < 0.01), while there was no significant difference between metal-ceramic appliances and natural teeth (p > 0.05). Although the worst findings were recorded for metal- + acrylic veneer crowns for PI, no significant difference existed between crowns of different material or non-abutment teeth (p > 0.05). There was statistically significant difference between abutments with metal + acrylic veneer crowns, full metal crowns, metal ceramic crowns and non-abutments for GI scores. Higher percentage of scores 0 and 1 was recorded for metal ceramic crowns and non-abutments and significantly higher percentage of scores 2 and 3 was recorded for metal + acrylic veneer crowns and full metallic crowns. Almost 50% of metal-ceramic abutments had no pathologic findings. Almost 30% of the patients needed replacement, or even some abutments to be extracted and therefore a new prosthodontic appliance.     

Influence of age and gender on radiomorphometric indices of the mandible in removable denture wearers

The objective of this study was to measure and assess a number of radiomorphometric indices of the mandibular removable denture wearers taking into account age and gender. In a total of 136 dental panoramic radiographs (DPR) the following parameters were evaluated: MI = thickness of the mandibular cortex below the mental foramen, AI = thickness at the antegonion and GI = thickness at gonion; PMI = MI/distance between the low border of the mandible and the low border of the mental foramen, and MCI = mandibular cortical index, based on the assessment of the 3 categories of cortical appearance due to the resorptive changes. There were 40 male patients (mean age 72.7; range 56 to 84 years) and 96 female patients (mean age 69.7; range 48 to 86 years). Patients were grouped according to age into three age groups (1 = less than 65 years; 2 = 65-75 years; 3 = more than 75 years). The results revealed that MI, PMI, AI and GI showed a general downward trend with age for the both sexes until 75 years of age when the mean values of GI, AI and MI begun to fall down sharply for females compared to males. The difference was significant between age groups and gender for GI and AI. Only two categories (C2 and C3) of MCI have been recognized in our study sample, due to the relatively old age groups of patients with removable dentures. However, significantly higher incidence of C3 existed in the oldest group of females.     

Characterization of mammalian eIF2A and identification of the yeast homolog

To begin the physical characterization of eukaryotic initiation factor (eIF) 2A, a translation initiation factor that binds Met-tRNA(i), tryptic peptides from rabbit reticulocyte eIF2A were analyzed to obtain amino acid sequence information. Sequences for 8 peptides were matched to three different expressed sequence tag clones. The sequence predicted for eIF2A is 585 amino acids. Matching of the cDNA sequence to the human genome revealed that the eIF2A mRNA is made up of 15 or 16 exons, and the gene is contained on chromosome 3. A homolog in Saccharomyces cerevisiae was identified, YGR054W, which is a non-essential gene. Hemagglutinin-tagged yeast eIF2A localizes on both 40 S and 80 S ribosomes. A knockout of both eIF2A and eIF5B yielded a "synthetically sick" yeast strain with a severe slow growth phenotype. The phenotype of this double mutant and the biochemical localization suggest that eIF2A participates in translation initiation. eIF2A does not appear to participate in re-initiation as the DeltaeIF2A strain shows the same level of GCN4 induction with amino acid starvation as seen in wild type yeast. The lack of any apparent phenotype in the DeltaeIF2A strain suggests that eIF2A functions in a minor pathway, perhaps internal initiation or in the translation of a small number of specific mRNAs.     

Metastasis of bronchogenic carcinoma to the thumb

Bone metastasis in the hand is rare. The etiology is quite different from that of metastasis to other bones; bronchogenic carcinoma is by far the most frequent case. Distal phalanges are mainly involved with irregular osteolysis and cortical destruction. Differential diagnosis of phalangeal metastasis includes osteomyelitis, rheumatoid arthritis and gout. The prognosis is always that of metastatic bronchial cancer with an average survival of three months. Treatment may involve distal digital amputation or antalgic radiotherapy. A case of bronchogenic carcinoma with metastasis to the thumb is presented. The metastasis was located in the distal phalanx of the left thumb. The primary tumor was located in the lung. Treatment consisted of amputation. The overall survival was five months.