Genetic analysis of delta helD and delta uvrD mutations in combination with other genes in the RecF recombination pathway in Escherichia coli: suppression of a ruvB mutation by a uvrD deletion

Helicase II (uvrD gene product) and helicase IV (helD gene product) have been shown previously to be involved in the RecF pathway of recombination. To better understand the role of these two proteins in homologous recombination in the RecF pathway [recBCsbcB(C) background, we investigated the interactions between helD, uvrD and the following RecF pathway genes: recF, recO, recN and ruvAB. We observed synergistic interactions between uvrD ant the recF, recN, recO and recG genes in both conjugational recombination and the repair of methylmethane sulfonate (MMS)-induced DNA damage. No synergistic interactions were detected between helD and the recF, recO and regN genes when conjugational recombination was analyzed. We did, however, detect synergistic interactions between helD and recF/recO in recombinational repair. Surprisingly, the uvrD deletion completely suppressed the phenotype of a ruvB mutation in a recBCsbcB(C) background. Both conjugational recombination efficiency and MMS-damaged DNA repair proficiency returned to wild-type levels in the deltauvrDruvB9 double mutant. Suppression of the effects of the ruvB mutation by a uvrD deletion was dependent on the recG and recN genes and not dependent on the recF/O/R genes. These data are discussed in the context of two "RecF" homologous recombination pathways operating in a recBCsbcB(C) strain background.     

Column Test Studies of Ochre Biofilm Formation in Geotextile Filters

This paper presents a study of ochre formation in geotextile filters. Ochre is a substance found sticking to solid surfaces of drainage systems that may cause clogging. The accumulation of such materials in a drainage system may have undesirable implications, such as a decrease in drainage flow capacity and an increase in soil pore pressure; instability of soil masses and retaining systems; and alteration of the direction of flow and development of pipes. Ochre formation is the result of microbial activities on iron compounds naturally found in seepage water. Even though geotextiles have been widely used in drainage systems, very few studies have considered ochre formation in the tests. Laboratory column filter tests were conducted to simulate the ochre formation process under similar conditions to those in geotechnical works and to assess the long-term performance of a geotextile. Ochre is most likely to be formed at an aerated/nonaerated interface in the filter. In order to obtain this condition, the underneath face of the filters was opened to the atmosphere. Three different types of geotextile—nonwoven polyester, nonwoven polypropylene, and woven polypropylene—were used in the tests. Control tests were also conducted with a sand filter to provide a basis for comparison. Hydraulic conductivity changes and iron retention within the permeameters were monitored throughout the tests. Ochre was formed under the presence of iron ions and iron bacteria. During the test period, the filters were not clogged to such an extent as to induce a global reduction in the permeability of the drainage system. Nevertheless, at the end of the tests a considerable amount of ochre was found in all filters and there was a significant variation in the permeability of the geotextile filters when tested in isolation. These studies may contribute to discovering whether ochre can be considered as a biofilm, evaluating the clogging potential and defining mitigating measures.     

Monitoring furosemide in racehorses participating in an EIPH program

We report a rare case of temporary and severe hypercalcemia: the patient, a 69-year-old woman, was admitted to Osaka City University Hospital on July 25, 1992, for severe hypercalcemia. The laboratory data on admission revealed severe hypercalcemia (14.9 mg/dl) and renal dysfunction with increased serum creatinine level (2.9 mg/dl). The urinary excretion of pyridinoline and deoxypyridinoline was increased, and serum levels of parathyroid hormone (PTH) and 1,25-dihydroxyvitamin D were decreased. The data suggested that increased bone resorption was a probable main factor in the development of the hypercalcemia. The development of hypercalcemia seemed to be of acute onset because of (1) her severe symptoms caused by hypercalcemia and (2) impaired renal function which was improved after normalization of serum calcium level. Combination therapy with saline infusion and furosemide was administered, and there was a gradual decrease and subsequent normalization of serum calcium level along with serum creatinine. Even 8 months after discontinuation of the therapy for hypercalcemia, the serum calcium level remained within the normal range. The measured values of serum factors which are suspected to have a hypercalcemic effect, such as PTH, parathyroid hormone-related peptide and the cytokines (interleukin-1 alpha, interleukin-1 beta, interleukin-2, interleukin-6 and tumor necrosis factor-alpha) were all within the normal range. In summary, the hypercalcemia in this patient was regarded to be a type of disequilibrium hypercalcemia due to a combination of increased bone resorption and decreased renal capacity to excrete calcium. Furthermore, since it was temporary and did not recur even in the absence of treatment, the hypercalcemia was concluded to have developed due to an imbalance in calcium regulation rather than as a result of organic disease.     

Radiation resistance and virulence of Listeria monocytogenes Scott A following starvation in physiological saline

The influence of starvation on the resistance of Listeria monocytogenes Scott A to electron beam irradiation in 0.85% (wt/vol) NaCl (saline) and in ground pork was investigated. Exponential- or stationary-phase cells (control) were grown at 35 degrees C in tryptic soy broth supplemented with 0.6% yeast extract. Washed cells were starved for 12 days in saline, and virulence of the pathogen was evaluated at 0, 8, and 12 days during starvation. Samples of saline and irradiation-sterilized ground pork, inoculated with control or starved cells, were irradiated at doses ranging from 0.0 to 2.5 kGy. L. monocytogenes survivors were determined by plating diluted samples of saline or pork on tryptic soy agar supplemented with 0.6% yeast extract and counting bacterial colonies following incubation (35 degrees C, 48 h). Virulence of starved cells and control was not significantly different (P > 0.05). Cells exhibited the highest radiation resistance at 8 days of starvation. Irradiation (0.5 kGy) in saline resulted in approximately 7.14, 5.55, and 2.38 log reduction in exponential, stationary, and starved cells, respectively. Irradiation of ground pork at 2.5 kGy reduced controls by approximately 6.0 log, whereas starved cells were reduced by only 3.8 log. Starved cells consistently exhibited higher irradiation D10-values than controls (P < 0.05). D10-values for exponential, stationary, and starved cells were 0.07, 0.09, and 0.21 kGy and 0.35, 0.42, and 0.66 kGy in saline and ground pork, respectively. These results indicate that starvation cross-protects L. monocytogenes Scott A against radiation inactivation and should be considered when determining this pathogen's irradiation D-value.     

The PROP1 2-base pair deletion is a common cause of combined pituitary hormone deficiency

Combined pituitary hormone deficiency (CPHD) has an incidence of approximately 1 in 8000 births. Although the proportion of familial CPHD cases is unknown, about 10% have an affected first degree relative. We have recently reported three mutations in the PROP1 gene that cause CPHD in human subjects. We report here the frequency of one of these mutations, a 301-302delAG deletion in exon 2 of PROP1, in 10 independently ascertained CPHD kindreds and 21 sporadic cases of CPHD from 8 different countries. Our results show that 55% (11 of 20) of PROP1 alleles have the 301-302delAG deletion in familial CPHD cases. Interestingly, although only 12% (5 of 42) of the PROP1 alleles of our 21 sporadic cases were 301-302delAG, the frequency of this allele (in 20 of 21 of the sporadic subjects given TRH stimulation tests) was 50% (3 of 6) and 0% (0 of 34) in the CPHD cases with pituitary and hypothalamic defects, respectively. Using whole genome radiation hybrid analysis, we localized the PROP1 gene to the distal end of chromosome 5q and identified a tightly linked polymorphic marker, D5S408, which can be used in segregation studies. Analysis of this marker in affected subjects with the 301-302delAG deletion suggests that rather than being inherited from a common founder, the 301-302delAG may be a recurring mutation.     

Local hyperthermia and SR 4233 enhance the antitumor effects of radioimmunotherapy in nude mice with human colonic adenocarcinoma xenografts

Local hyperthermia and the hypoxic cytotoxin SR 4233 were administered to nude mice with 693 +/- 47 mm3 (mean +/- SE) s.c. HCT-8 human colonic adenocarcinoma xenografts in an attempt to enhance the antitumor effects of radioimmunotherapy. Biodistribution studies revealed preferential binding of NR-Lu-10, a murine monoclonal antibody, to the tumors compared with an isotype-matched control antibody, CCOO16-3.A single injection of 25 microCi 90Y-NR-Lu-10 significantly inhibited tumor growth (control versus 90Y-NR-Lu-10: P = 0.048). The administration of hyperthermia at 41.5 degrees C for 1 h immediately following the injection of 111In-labeled NR-Lu-10 up-regulated tumor-associated antigen expression and increased antibody uptake in the tumors by 73% (P = 0.001) without significantly affecting antibody uptake in normal tissues. However, the heat treatment did not produce a more homogeneous distribution of the antibodies in the tumors and did not significantly enhance the tumor growth delay produced by 90Y-NR-Lu-10 (P = 0.07). The administration of local hyperthermia at 43.0 degrees C for 1 h, on the other hand, had direct cytotoxic effects (P = 0.03) and enhanced the tumor growth delay produced by 90Y-NR-Lu-10 (P = 0.01). SR 4233 also enhanced the tumor growth delay produced by 90Y-NR-Lu-10 (P = 0.03). The greatest antitumor effects were observed when both hyperthermia at 43.0 degrees C and SR 4233 were administered in combination with 90Y-NR-Lu-10 (P = 0.002). No toxicity was produced by the local hyperthermia, and the only toxicities produced by 90Y-NR-Lu-10 and SR 4233 were neutropenia and weight loss.     

Survival of Listeria monocytogenes and Escherichia coli O157:H7 during sauerkraut fermentation

Sauerkraut was produced from shredded cabbage, as is typical in the United States, and from whole head cabbages, which is a traditional process in parts of Eastern Europe. The sauerkraut was inoculated with five strain mixtures of Escherichia coli O157:H7 and Listeria monocytogenes, and the populations of these bacteria, as well as lactic acid bacteria, pH, and titratable acidity, were monitored over the course of fermentation. Fermentation variables were temperature (18 and 22 degrees C) and salt concentration (1.8, 2.25, and 3%). For most of the analyses, the type of cabbage processing was a significant factor, although within cabbage type, neither salt nor fermentation temperature had significant effects. The final pH of the whole-head sauerkraut was lower than the shredded sauerkraut, but the titratable acidity was significantly higher in the shredded sauerkraut. E. coli O157:H7 and L. monocytogenes persisted in the brines for most of the fermentation, although at the end of the fermentations (15 days for shredded, 28 days for whole head), neither pathogen had detectable populations. E. coli populations decreased more rapidly in the shredded sauerkraut even though the pH was higher because of the higher total acidity in the shredded sauerkraut. Acid-tolerant strains of E. coli and L. monocytogenes were isolated from both shredded and whole-head sauerkraut at different salt concentrations and temperatures after 15 days of fermentation and could be detected at 35 days in the wholehead sauerkraut.     

Water pressure effectively reduces Salmonella enterica serovar enteritidis on the surface of raw almonds

The effects of continuous high hydrostatic water pressure treatments (414 and 483 MPa with a holding time of 6 min) on the viability of two strains of Salmonella enterica serovar Enteritidis (PT 9c and PT 30) inoculated onto raw almonds were evaluated at 25 degrees C. The concentrations of both Salmonella Enteritidis isolates were reduced to undetectable levels when the almonds were directly suspended in water, pressurized at 414 MPa and 25 degrees C for 6 min, and then dried at 115 degrees C for 25 min. When the almonds were pressurized at 414 MPa and dried at 55 degrees C for 5 min, a more than 3.58-log reduction was achieved for both isolates. Increasing the drying temperature to 65 degrees C resulted in a more than 3.96-log reduction for both isolates. Increasing the water pressure to 483 MPa further decreased the Salmonella Enteritidis concentration on the surface of the raw almonds. When the almonds were treated at 414 MPa, the decimal reduction times were less than 35 s for both isolates. High hydrostatic water pressure treatment of certain dry foods appears to be feasible when the food is suspended directly in the pressurizing medium (water).     

Influence of four retail food service cooling methods on the behavior of Clostridium perfringens ATCC 10388 in turkey roasts following heating to an internal temperature of 74 degrees C

The influence of four food service cooling methods (CM) on growth of Clostridium perfringens ATCC 10388 in cooked turkey roasts was evaluated. Raw whole turkey roasts were inoculated with C. perfringens spores (approximately 4.23 log CFU per roast), vacuum packaged, and heated to an internal temperature of 74 degrees C. The cooked roasts were cooled as follows: whole roast cut into four quarters and held at 4 degrees C (CM1); whole roast held in a blast chiller (CM2); whole roast loosely wrapped and held at 4 degrees C (CM3); and whole roasts (three per bag) held at 4 degrees C (CM4). The roasts were analyzed for C. perfringens using Shahidi-Ferguson perfringens agar and anaerobic incubation (37 degrees C, 24 h). None of the cooling methods met the amended 2001 U.S. Food and Drug Administration Food Code guidelines for safe cooling of potentially hazardous foods. Times taken for roasts to cool from 57 to 21 degrees C using CM1, CM2, CM3, and CM4 were 2.27, 3.11, 6.22, and 8.71 h, respectively. Times taken for roasts (21 degrees C) to reach 5 degrees C ranged from 6.33 (CM1) to 19.45 h (CM4). Based on initial numbers of C. perfringens, no growth occurred in roasts cooled by CM1 or CM2, whereas numbers increased by 1.5 and 4.0 log in whole roasts cooled via CM3 and CM4, respectively. These findings indicate that certain food service cooling methods for whole cooked turkey roasts may result in proliferation of C. perfringens and increase the risk of foodborne illness by this pathogen.     

Combining pediocin with postpackaging irradiation for control of Listeria monocytogenes on frankfurters

Frankfurters, in 1-link, 5-link, or 10-link packages, were surface inoculated with a five-strain mixture of Listeria monocytogenes (3.40 or 5.20 log CFU/g) after treatment with 3,000 arbitrary units (AU) or 6,000 AU of pediocin (in ALTA 2341) per link. The frankfurters were vacuum packaged, after which the 1-link and 5-link packages were irradiated at 1.2 or 2.3 kGy and the 10-link packages were irradiated at 1.4 or 3.5 kGy. L. monocytogenes was enumerated following the treatments. Selected treatments were subsequently evaluated during storage at 4, 10, and 25 degrees C for up to 12 weeks. Combination of pediocin with postpackaging irradiation at 1.2 kGy or more was necessary to achieve a 50% reduction of L. monocytogenes on frankfurters in 1-link or 5-link packages. The combination of 6,000 AU of pediocin and irradiation at 2.3 kGy or more was effective in all package sizes for inhibition of the pathogen for 12 weeks at 4 or 10 degrees C. There was a synergistic effect between pediocin and irradiation for inhibition of L. monocytogenes. Storage at 4 degrees C enhanced the antilisterial effects of the treatment combinations, with little or no growth of the pathogen in 1-link or 5-link packages during 12 weeks of storage. In general, these treatments did not affect the sensory quality of frankfurters.