Plasmid pGA1 from Corynebacterium glutamicum codes for a gene product that positively influences plasmid copy number

The complete nucleotide sequence (4,826 bp) of the cryptic plasmid pGA1 from Corynebacterium glutamicum was determined. DNA sequence analysis revealed four putative coding regions (open reading frame A [ORFA], ORFA2, ORFB, and ORFC). ORFC was identified as a rep gene coding for an initiator of plasmid replication (Rep) according to the high level of homology of its deduced amino acid sequence with the Rep proteins of plasmids pSR1 (from C. glutamicum) and pNG2 (from Corynebacterium diphtheriae). This function was confirmed by deletion mapping of the minimal replicon of pGA1 (1.7 kb) which contains only ORFC. Deletion derivatives of pGA1 devoid of ORFA exhibited significant decreases in the copy number in C. glutamicum cells and displayed segregational instability. Introduction of ORFA in trans into the cells harboring these deletion plasmids dramatically increased their copy number and segregational stability. The ORFA gene product thus positively influences plasmid copy number. This is the first report on such activity associated with a nonintegrating bacterial plasmid. The related plasmids pGA1, pSR1, and pNG2 lacking significant homology with any other plasmid seem to be representatives of a new group of plasmids replicating in the rolling-circle mode.     

Influence of total surface area of core material on yield of deposited coacervate

The neuronal cytoskeleton is one of the most profoundly altered organelles in late life neuro-degenerative disorders that are characterized by progressive impairments in cognitive abilities. The elucidation of the protein building blocks of these organelles as well as advances in understanding how these proteins become altered in Alzheimer's disease (AD) and other less common dementing illnesses, i.e., diffuse Lewy body disease (DLBD) or the Lewy body variant of AD (LBVAD), will provide insights into the molecular basis of these disorders. Within, we review evidence that normal adult human tau is abnormally phosphorylated and converted into the subunits of AD paired helical filaments (PHFs), and that Lewy bodies (LBs) represent accumulation of altered neurofilament (NF) triplet subunits. Although the precise biological consequences of PHF and LB formation in neurons is unknown, growing evidence suggests that the formation of PHFs and LBs from normal neuronal cytoskeletal proteins could have deleterious effects on neuronal function and survival. Finally, insights into the composition of PHFs and LBs could lead to the development of novel strategies for the timely and accurate diagnosis of AD, DLBD and the LBVAD.     

Predictive equation for acquisition of hepatitis B in hospital workers in a general hospital

A cross-sectional study was performed to obtain risk factors for hepatitis B disease, HBsAg carriers and immunised personnel, among 2470 workers in a general hospital in Madrid, Spain. The data obtained were analyzed with multiple logistic regression to obtain beta coefficients for variables. The results of the analysis show that being a nurse or being regularly exposed to blood are the most important risk factors for hepatitis B acquisition. The length of time working at the same job activity was also a risk factor. The resulting beta coefficients allow the construction for a hepatitis non-immunised, HBsAg carrier and immunised HBV status, which can select subjects for a hepatitis B vaccination program.     

Processing of Yttria-Alumina Coated Silicon Nitride Slips by Slip Casting

Si₃N₄powders coated with 6 wt% Y₂O₃and 4 wt% Al₂O₃were prepared by coprecipitation. The resulting powders were dispersed in water at different pH values and with addition of various amounts of ammonium polyacrylate (NH₄PA) to produce 32 vol% slips. The influence of the amount of NH₄PA solution added and pH on the rheological properties of 32 vol% coated Si₃N₄slips were studied. In addition, the sintered density of cast samples was determined and related to the degree of slip dispersion. The adsorption of the NH₄PA on the coated particle surface was rather high and the surface became saturated near 0.86 mg/m²at pH 9.2. High NH₄PA concentrations (1.7–3 wt%) were necessary to obtain well dispersed 32 vol% coated Si₃N₄slips at pH 9.2. The best stabilization was obtained with the addition of 2.3 wt% NH₄PA; in this condition, the viscosity reached a minimum value of 35 mPa.s at 100 s^–1. The slip viscosity increased with increasing pH from 9.2 to 10.2. Slips with low viscosities gave a more dense packing of cast samples and consequently higher sintered density values.     

Generation of Technology-Independent Retargetable Analog Blocks

This paper introduces a complete methodology for retargeting of analog cells to different sets of specifications. This methodology is technology-independent, thus allowing the migration, from one technology to another, of the circuit under retargeting. Careful integration of the device sizing and layout generation tasks via the incorporation of layout constraints during the sizing process allows to generate fully functional designs in a few minutes. The methodology is illustrated via the retargeting of a fully-differential Miller-compensated two-stage operational amplifier for a new set of specifications and two different technological processes.An erratum to this article can be found at     

PCR-based procedures in detection and DNA-fingerprinting of Salmonella from samples of animal origin

The usefulness of selected PCR-protocols for the detection of Salmonella in 117 samples of animal origin (17 raw minced meat, 27 raw chicken meat, 8 raw sausages, and 25 egg samples, as well as 18 poultry faecal, and caecal swabs samples) and DNA-fingerprinting typing is shown. To establish an accurate PCR-procedure for Salmonella detection the following parameters were evaluated: two pre-PCR concentration procedures, centrifugation and immunomagnetic separation (IMS) using Dynabeads anti- Salmonella; the specificity and sensitivity of 10 sets of primers; and different conditions of the amplification reaction. In light of the results obtained from the use of PCR-based procedures alone or in combination with conventional methods, the following findings can be underlined: First, IMS is more efficient than centrifugation in the recovery of Salmonella. Second, the selected IMS/PCR-detection protocol is less time-consuming (45 h) than the IMS/culture procedure (90 h), and a good concordance between them was found when the Kappa coefficient was calculated (0·87). Third, PCR-ribotyping technique showed a very low discrimination power, being able to differentiate only three profiles. Fourth, RAPD technique using specific primers supports previous works in which it was proposed as a simple and useful tool for discriminating isolates between and within serotypes. Fifth, The efficiency, rapidity, and flexibility of the PCR-protocols applied were high, and they can be performed using two PCR-programs and the same basic equipment.