Evolution ofE.coli tRNATrp

Earlier studies (1) have shown there are direct correlations between the hydrophobicity ranking of most amino acids and their anticodonic nucleotides. However, four anticodonic assignments, i.e. those for Trp, Tyr, Ile and the XGA anticodons for Ser, did not correlate. It was our proposal that this failure to correlate was due to the fact that these assignments were made late, relative to the bulk of the assignments, in evolution through the mutation of existing tRNAs. We have shown (2) thatE. coli tRNA^{Ile 1} and tRNA^{Ile 2} were likely derived from tRNA^{Val 1} and tRNA^Lys respectively andE. coli tRNA^Tyr was possibly derived fromE. coli 5s rRNA or a common precursor with 5s rRNA (3). The fact that quite high homologies were observed in these comparisons is consistent with the late evolution of the tRNAs in question. We now examine the evolution ofE. coli tRNA^Trp by comparing its homology with otherE. coli tRNAs. The data suggest a possible evolutionary relationship withE. coli tRNA^Gly or tRNA^Arg. The data support the idea of the late assignment of anticodons to Trp.Deceased.     

The structural periodicity ofE. coli ribosomal proteins

It is established that the sequences of all different proteins fromE. coli ribosome as well as two protein biosynthesis initiation factors, two ribosome-associated DNA-binding proteins, and the elongation factor EF-Tu from the same source possess a periodicity expressed more weakly and different from that found earlier for a number of proteins representatives of 18 superfamilies. The statistical significance of the periodicity observed was checked by comparing the area below the periodicity curve of every protein examined with that of computer generated sequences having the same amino acid composition and length. The results concerning the proteins from small and large ribosomal subunit are compared. The conclusions support and supplement the concept about the presence of a trend in protein molecular evolution from universal (Gly, Ala) to specialized (Phe, Tyr, Trp, Cys) amino acids.     

On the critical behaviour of simple epidemics.

We show how ideas and models which were originally introduced to gain an understanding of critical phenomena can be used to interpret the dynamics of epidemics of communicable disease in real populations. Specifically, we present an analysis of the dynamics of disease outbreaks for three common communicable infections from a small isolated island population. The strongly fluctuating nature of the temporal incidence of disease is captured by the model, and comparisons between exponents calculated from the data and from simulations are made. A forest-fire model with sparks is used to classify the observed scaling dynamics of the epidemics and provides a unified picture of the epidemiology which conventional epidemiological analysis is unable to reproduce. This study suggests that power-law scaling can emerge in natural systems when they are driven on widely separated time-scales, in accordance with recent analytic renormalization group calculations.     

On the rate of water exchange across the surface of animal cells

The two methods for determination of water permeability, the osmometric method, and the isotope exchange method are discussed, particularly with respect to the sources of error. In the latter method the exchange across the surface barrier is generally so fast that correction for the diffusion in the cytoplasm is required. The presence of cortical layers of considerable mechanical strength may often invalidate results obtained with the former method. In agreement with views advanced previously in the literature, it is concluded that the permeability coefficients obtained by the two different methods only under very special conditions are identical. However, the ratio cannot be expected to deviate much from unity, and it is shown that the large ratios reported in some cases do not represent the ratio between two permeability coefficients.     

Inactivation of polioviruses and coxsackieviruses in surface water.

Inactivation rates of polioviruses 1 and 3 and coxsackieviruses A-13 and B-1 were determined in situ in the Rio Grande in southern New Mexico, using membrane dialysis chambers. Inactivation of the viruses was exponential, and the rates of inactivation were apparently affected principally by the water temperature. Stability of the viruses in river water differed, with poliovirus 1 and coxsackie B-1 being most stable. Typically 1-log reductions of infectivity at water temperatures ranging between 23 and 27 degrees C required 25 h for poliovirus 1, 19 h for poliovirus 3, and 7 h for coxsackie virus A-13. At water temperatures of 4 to 8 degrees C, the log reduction times for poliovirus 1 and coxsackievirus B-1 were 46 and 58 h, respectively. Results obtained with labeled poliovirus 1 and coxsackievirus B-1 and with infectious ribonucleic acid indicate that inactivation was due to damage to viral ribonucleic acid. Virus-inactivation rates were also affected by heat sterilization of river water, indicating the presence of a heat-labile or volatile inactivating factor. The inactivating factor in Rio Grande water was apparently present at a constant concentration over a 1-year period.     

Inactivation of polioviruses and coxsackieviruses in surface water.

Inactivation rates of polioviruses 1 and 3 and coxsackieviruses A-13 and B-1 were determined in situ in the Rio Grande in southern New Mexico, using membrane dialysis chambers. Inactivation of the viruses was exponential, and the rates of inactivation were apparently affected principally by the water temperature. Stability of the viruses in river water differed, with poliovirus 1 and coxsackie B-1 being most stable. Typically 1-log reductions of infectivity at water temperatures ranging between 23 and 27 degrees C required 25 h for poliovirus 1, 19 h for poliovirus 3, and 7 h for coxsackie virus A-13. At water temperatures of 4 to 8 degrees C, the log reduction times for poliovirus 1 and coxsackievirus B-1 were 46 and 58 h, respectively. Results obtained with labeled poliovirus 1 and coxsackievirus B-1 and with infectious ribonucleic acid indicate that inactivation was due to damage to viral ribonucleic acid. Virus-inactivation rates were also affected by heat sterilization of river water, indicating the presence of a heat-labile or volatile inactivating factor. The inactivating factor in Rio Grande water was apparently present at a constant concentration over a 1-year period.     

Identification of Immunogenic Salmonella enterica Serotype Typhi Antigens Expressed in Chronic Biliary Carriers of S. Typhi in Kathmandu,Nepal

Background

Salmonella enterica serotype Typhi can colonize and persist in the biliary tract of infected individuals, resulting in a state of asymptomatic chronic carriage. Chronic carriers may act as persistent reservoirs of infection within a community and may introduce infection to susceptible individuals and new communities. Little is known about the interaction between the host and pathogen in the biliary tract of chronic carriers, and there is currently no reliable diagnostic assay to identify asymptomatic S. Typhi carriage.

Methodology/Principal Findings

To study host-pathogen interactions in the biliary tract during S. Typhi carriage, we applied an immunoscreening technique called in vivo-induced antigen technology (IVIAT), to identify potential biomarkers unique to carriers. IVIAT identifies humorally immunogenic bacterial antigens expressed uniquely in the in vivo environment, and we hypothesized that S. Typhi surviving in the biliary tract of humans may express a distinct antigenic profile. Thirteen S. Typhi antigens that were immunoreactive in carriers, but not in healthy individuals from a typhoid endemic area, were identified. The identified antigens included a number of putative membrane proteins, lipoproteins, and hemolysin-related proteins. YncE (STY1479), an uncharacterized protein with an ATP-binding motif, gave prominent responses in our screen. The response to YncE in patients whose biliary tract contained S. Typhi was compared to responses in patients whose biliary tract did not contain S. Typhi, patients with acute typhoid fever, and healthy controls residing in a typhoid endemic area. Seven of 10 (70%) chronic carriers, 0 of 8 bile culture-negative controls (0%), 0 of 8 healthy Bangladeshis (0%), and 1 of 8 (12.5%) Bangladeshis with acute typhoid fever had detectable anti-YncE IgG in blood. IgA responses were also present.

Conclusions/Significance

Further evaluation of YncE and other antigens identified by IVIAT could lead to the development of improved diagnostic assays to identify asymptomatic S. Typhi carriers.     

Influence of oxygen supply in the production of biomass ofE. coli cells immobilized in K-carrageenan gel

Summary Cells of a recombinantEscherichia coli, immobilized in K-carrageenan, show improved distribution of growth in the gel when cultured in conditions of high oxygen tension. The production of both biomass and protein codified by recombinant DNA is increased.     

Optimization of Salmonella Typhi biofilm assay on polypropylene microtiter plates using response surface methodology

The objective of this study was to develop an optimized assay for Salmonella Typhi biofilm that mimics the environment of the gallbladder as an experimental model for chronic typhoid fever. Multi-factorial assays are difficult to optimize using traditional one-factor-at-a-time optimization methods. Response surface methodology (RSM) was used to optimize six key variables involved in S. Typhi biofilm formation on cholesterol-coated polypropylene 96-well microtiter plates. The results showed that bile (1.22%), glucose (2%), cholesterol (0.05%) and potassium chloride (0.25%) were critical factors affecting the amount of biofilm produced, but agitation (275 rpm) and sodium chloride (0.5%) had antagonistic effects on each other. Under these optimum conditions the maximum OD reading for biofilm formation was 3.4 (λ_600 nm), and the coefficients of variation for intra-plate and inter-plate assays were 3% (n?=?20) and 5% (n?=?8), respectively. These results showed that RSM is an effective approach for biofilm assay optimization.     

Occurrence of Giardia and Cryptosporidium spp. in surface water supplies.

Giardia and Cryptosporidium levels were determined by using a combined immunofluorescence test for source waters of 66 surface water treatment plants in 14 states and 1 Canadian province. The results showed that cysts and oocysts were widely dispersed in the aquatic environment. Giardia spp. were detected in 81% of the raw water samples. Cryptosporidium spp. were found in 87% of the raw water locations. Overall, Giardia or Cryptosporidium spp. were detected in 97% of the raw water samples. Higher cyst and oocyst densities were associated with source waters receiving industrial or sewage effluents. Significant correlations were found between Giardia and Cryptosporidium densities and raw water quality parameters such as turbidity and total and fecal coliform levels. Statistical modeling suggests that cyst and oocyst densities could be predicted on the basis of watershed and water quality characteristics. The occurrence of high levels of Giardia cysts in raw water samples may require water utilities to apply treatment beyond that outlined in the Surface Water Treatment Rule of the U.S. Environmental Protection Agency.     

Occurrence of Giardia and Cryptosporidium spp. in surface water supplies.

Giardia and Cryptosporidium levels were determined by using a combined immunofluorescence test for source waters of 66 surface water treatment plants in 14 states and 1 Canadian province. The results showed that cysts and oocysts were widely dispersed in the aquatic environment. Giardia spp. were detected in 81% of the raw water samples. Cryptosporidium spp. were found in 87% of the raw water locations. Overall, Giardia or Cryptosporidium spp. were detected in 97% of the raw water samples. Higher cyst and oocyst densities were associated with source waters receiving industrial or sewage effluents. Significant correlations were found between Giardia and Cryptosporidium densities and raw water quality parameters such as turbidity and total and fecal coliform levels. Statistical modeling suggests that cyst and oocyst densities could be predicted on the basis of watershed and water quality characteristics. The occurrence of high levels of Giardia cysts in raw water samples may require water utilities to apply treatment beyond that outlined in the Surface Water Treatment Rule of the U.S. Environmental Protection Agency.     

Yersinia spp. in surface water in San Luis,Argentina

Yersinia spp. was examined in three rivers and two lakes located in the Province of San Luis, Argentina, over a 1-year period. Water samples were concentrated either by Moore's gauze technique or by filtering through diatomaceous earth. Five enrichment media: yeast extract-Bengal rose broth (YER) with bile-oxalate-sorbose broth (BOS); 67 mmol/L phosphatebuffered saline (pH 7.6; PBS); PBS enriched with 1% mannitol and 1% peptone (PBSMP); PBS with lyzed 0.5% sheep blood (PBSB); Wauters broth (W); and five plating media: Mac Conkey agar (MC);Salmonella—Shigella agar (SS); 5% sheep blood agar (BA); lactose-sucrose-urea agar (LSU) and irgasan-novobiocin agar (IN) were used. The following strains were isolated:Y. intermedia B1 O:4,32–4,33 Lis Xz (four strains),Y. intermedia B1 O:57 Lis Xo (one strain),Y. intermedia B2 0:57 Lis Xo (one strain),Y. enterocolitica B1 O:10–34 Lis Xz (one strain), andY. frederiksenii undetermined biovar, O:16–16,29 Lis Xz (two strains). The incidence of isolation ofYersinia spp. was 7.14%. YER-BOS proved to be the best enrichment method since it allowed the highest recovering ofYersinia spp. strains. Among plating media, the best results were obtained with MC. Apparently, the isolation ofYersinia spp. can be related to environmental variables such as temperature differences between cold and warm seasons. Negative results obtained during virulence assays suggest that isolated strains lack the pathogenic potential against man.     

Upregulation ofE. coli 38kDa proteins induced by glutaraldehyde and formaldehyde

Exposure of the W3110 strain ofEscherichia coli K12 to low concentrations of glutaraldehyde or formaldehyde results in an unusual pattern of protein expression, as determined by high-resolution, two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). A decline in total protein synthesis is accompanied by the upregulation of three proteins of approximate molecular weight 38 kDa. In the presence of 0.1 mM glutaraldehyde this response occurs within the first 5 min of incubation, and with 0.1 mM formaldehyde, within the first 30 min of incubation. The 38 kDa proteins continue to be expressed at high levels until cell death. Comparison of our 2-D PAGE patterns withE. coli gene-protein and plasmid indexes indicates that one of the proteins may be the major gene product of thepyrC locus. This pattern of protein synthesis may indicate a novelE. coli stress response.     

Reproduction of bacteriophage T6 and T7 in auxotrophic mutants ofE. coli B

Summary Phage T₆ and T₇ did not reproduce in mutants ofE. coli auxotrophic for the amino acids methionine, leucine, serine, proline, histidine, tyrosine, phenylalanine and tryptophane unless these amino acids were present in the medium. Very small amounts of these auxotrophic factors (0.01 γ/ml) are able to support phage multiplication. Much care has to be taken to avoid impurities in the medium and to deplete the endogenous reserves of the host cells.