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Biomedical subjects

B Ray

Publications and source records attributed to B Ray.

At least 37 records · Page 2Linked to original sources
PubMed2000

Interactions of high hydrostatic pressure, pressurization temperature and pH on death and injury of pressure-resistant and pressure-sensitive strains of foodborne pathogens.

The objective of this study is to determine the interactions between high hydrostatic pressure, pressurization temperature, time and pH during pressurization on death and injury of pressure-resistant and pressure-sensitive strains of four foodborne pathogens: Staphylococcus aureus 485 and 765, Listeria ,monocytogenes CA and OH2, Escherichia coli O157:H7 933 and 931, Salmonella enteritidis FDA and Salmonella typhimurium E21274. Among these strains S. aureus 485, L. monocytogenes CA, E. coli O157:H7 933 and S. enteritidis FDA were reported to be more pressure-resistant than the respective strain of the same species (Alpas et al., 1999). In general, viability loss of all pathogens was enhanced significantly as the level of pressure and temperature were increased (P < 0.05). All the strains except S. aureus 485 demonstrated more than 8 log cycle reduction when pressurized at 345 MPa at 50 degrees C for 5 min. This strain seemed to be the most pressure-resistant strain within the conditions of the study. Pressurization in the presence of either citric or lactic acid increased the viability loss by an additional 1.2-3.9 log cycles at pH 4.5 for both acids at 345 MPa. This study has indicated that high hydrostatic pressure applied in conjunction with mild heat and acidity can be an effective method for inactivating pressure-resistant and pressure-sensitive strains of four foodborne pathogens in organic acid solutions. This combination treatment indicates possible pressure pasteurization applications to liquid foods that have low pH. reserved.

Analysis of Variance↗
PubMed2000

Vibrio parahaemolyticus infections in the United States, 1973-1998.

Vibrio parahaemolyticus infections are associated with consumption of raw or undercooked shellfish, contaminated food, and exposure of wounds to warm seawater. Foodborne outbreaks and sporadic infections from Vibrio species in 4 Gulf Coast states are reported routinely to the Centers for Disease Control and Prevention (CDC). Between 1988 and 1997, 345 sporadic V. parahaemolyticus infections were reported: 59% were gastroenteritis, 34% were wound infections, 5% were septicemia, and 2% were from other exposures. Forty-five percent of patients suffering from these conditions were hospitalized for their infections, and 88% of persons with acute gastroenteritis reported having eaten raw oysters during the week before their illness occurred. Between 1973 and 1998, 40 outbreaks of V. parahaemolyticus infections were reported to the CDC, and these outbreaks included >1000 illnesses. Most of these outbreaks occurred during the warmer months and were attributed to seafood, particularly shellfish. The median attack rate among persons who consumed the implicated seafood was 56%. To prevent V. parahaemolyticus infections, persons should avoid consumption of raw or undercooked shellfish and exposure of wounds to seawater.

Centers for Disease Control and Prevention, U.S.↗
PubMed2000

Consequences of placing an intramolecular crosslink in myosin S1.

This paper describes the placement of a crosslinking agent (dibromobimane) between two thiols (Cys-522 and Cys-707) of a fragment, "S1," of the motor protein, myosin. It turns out that fastening the first anchor of the crosslinker is easy and rapid, but fastening the second anchor (Cys-522) is very temperature dependent, taking 30 min at room temperature but about a week on ice. Moreover, crystallography taken at 4 degrees C would seem to predict that the linkage is impossible, because the span of the crosslinking agent is much less than the interthiol distance. The simplest resolution of this seeming paradox is that structural fluctuations of the protein render the linkage increasingly likely as the temperature increases. Also, measurements of the affinity of MgADP for the protein, as well as the magnetic resonance of the P-atoms of the ADP once emplaced, suggest that binding the first reagent anchor to Cys-707 initiates an influence that travels to the rather distant ADP-binding site, and it is speculated what this "path of influence" might be.

Adenosine Diphosphate↗
PubMed2000

A survey of stool culturing practices for vibrio species at clinical laboratories in Gulf Coast states.

Non-cholera Vibrio infections are an important public health problem. Non-cholera Vibrio species usually cause sporadic infections, often in coastal states, and have also caused several recent nationwide outbreaks of gastroenteritis in the United States. We report a survey of laboratory stool culturing practices for Vibrio among randomly selected clinical laboratories in Gulf Coast states (Alabama, Florida, Louisiana, Mississippi, and Texas). Interviews conducted with the microbiology supervisors of 201 clinical laboratories found that 164 (82%) received stool specimens for culture. Of these, 102 (62%) of 164 processed stool specimens on site, and 20 (20%) of these 102 laboratories cultured all stool specimens for Vibrio, indicating that at least 34,463 (22%) of 152, 797 stool specimens were cultured for Vibrio. This survey suggests that despite an increased incidence of non-cholera Vibrio infections in Gulf Coast states, a low percentage of clinical laboratories routinely screen all stool specimens, and fewer than 25% of stool specimens collected are routinely screened for non-cholera Vibrio.

Alabama↗
PubMed1999

The pediocin AcH precursor is biologically active.

The properties of the pediocin AcH precursor, prepediocin AcH, have been studied to gain insight into how producer cells may protect themselves from the activity of intracellular prebacteriocins. The native 62-amino-acid precursor and the 44-amino-acid mature species were expressed in Escherichia coli host strains that lack the leader peptide processing enzyme, PapD. Both forms inhibited the growth of the test bacterium Listeria innocua Lin11, indicating that the native precursor is biologically active. The two species also were synthesized in the context of maltose-binding protein chimeric proteins to facilitate the measurement of their relative specific activities. The chimeric form of the precursor was approximately 80% as active as the chimeric mature species. Of relevance to cell protection and pediocin AcH production, it was determined that the precursor is strongly susceptible to inactivation by reducing agents and to degradation by chymotrypsin and endogenous E. coli proteases. Taken together, the results indicate that the activity of prepediocin AcH may have to be controlled prior to secretion to prevent toxicity to the host. Perhaps producer cells avoid membrane damage by maintaining the precursor in a reduced inactive state or by degrading molecules whose secretion is delayed.

ATP-Binding Cassette Transporters↗
PubMed1999

Variation in resistance to hydrostatic pressure among strains of food-borne pathogens.

Among food-borne pathogens, some strains could be resistant to hydrostatic pressure treatment. This information is necessary to establish processing parameters to ensure safety of pressure-pasteurized foods (N. Kalchayanand, A. Sikes, C. P. Dunne, and B. Ray, J. Food Prot. 61:425-431, 1998). We studied variation in pressure resistance among strains of Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella species at two temperatures of pressurization. Early-stationary-phase cells in 1% peptone solution were pressurized at 345 MPa either for 5 min at 25 degrees C or for 5, 10, or 15 min at 50 degrees C. The viability loss (in log cycles) following pressurization at 25 degrees C ranged from 0.9 to 3.5 among nine L. monocytogenes strains, 0.7 to 7.8 among seven S. aureus strains, 2.8 to 5.6 among six E. coli O157:H7 strains, and 5.5 to 8.3 among six Salmonella strains. The results show that at 25 degrees C some strains of each species are more resistant to pressure than the others. However, when one resistant and one sensitive strain from each species were pressurized at 345 MPa and 50 degrees C, the population of all except the resistant S. aureus strain was reduced by more than 8 log cycles within 5 min. Viability loss of the resistant S. aureus strain was 6.3 log cycles even after 15 min of pressurization. This shows that strains of food-borne pathogens differ in resistance to hydrostatic pressure (345 MPa) at 25 degrees C, but this difference is greatly reduced at 50 degrees C. Pressurization at 50 degrees C, in place of 25 degrees C, will ensure greater safety of foods.

Bacteria↗
PubMed1998

Human male germ cell tumor resistance to cisplatin is linked to TP53 gene mutation.

Male germ cell tumors (GCTs) are uniquely sensitive to cisplatin-based chemotherapy, with more than 90% of newly diagnosed cases cured. The underlying cause for resistance to treatment in 20-30% of metastatic lesions remains to be identified. Unlike other solid tumors, no mutations in the TP53 gene have been identified to date in random panels of GCT specimens, which could account for the exquisite sensitivity of these tumors to genotoxic insult. However, in a panel of resistant GCTs that did either not respond to cisplatin-based chemotherapy or subsequently relapsed and resulted in the death of the patient, we have now identified a subset of tumors to contain TP53 mutations within exons 6-9. A cell line derived from one of these tumors (228A) displayed the same TP53 mutation as the tumor specimen, expressed only mutant TP53 mRNA, and exhibited a relative resistance to cisplatin in vitro in comparison to a cell line (218A) derived from a responsive tumor with wild-type TP53. The resistant cell line displayed a much reduced apoptotic cell death and did not exhibit an induction of transcription of the p53-responsive genes WAF1 and MDM2 following cisplatin treatment, compared to that observed in the sensitive cell line. The levels of bax, an agonist of apoptosis, were found to be reduced in the resistant cell line. The simplest explanation for the resistance of this subset of GCTs that are resistant to cisplatin-based chemotherapy, is the inability of the cells to mount an apoptotic response following exposure due to a functionally inactivating mutation in the TP53 gene.

Antineoplastic Agents↗
PubMed1998

Crayfish: a newly recognized vehicle for vibrio infections.

We conducted a 1-year case-control study of sporadic vibrio infections to identify risk factors related to consumption of seafood products in two coastal areas of Louisiana and Texas. Twenty-six persons with sporadic vibrio infections and 77 matched controls were enrolled. Multivariate analysis revealed that crayfish (P < 0.025) and raw oysters (P < 0.009) were independently associated with illness. Species-specific analysis revealed an association between consumption of cooked crayfish and Vibrio parahemolyticus infection (OR 9.24, P < 0.05). No crayfish consumption was reported by persons with V. vulnificus infection. Although crayfish had been suspected as a vehicle for foodborne disease, this is the first time to our knowledge that consumption of cooked crayfish has been demonstrated to be associated with vibrio infection.

Animals↗
PubMed1998

The role of Gulf Coast oysters harvested in warmer months in Vibrio vulnificus infections in the United States, 1988-1996. Vibrio Working Group.

Vibrio vulnificus infections are highly lethal and associated with consumption of raw shellfish and exposure of wounds to seawater. V. vulnificus infections were reported to the Centers for Disease Control and Prevention from 23 states. For primary septicemia infections, oyster trace-backs were performed and water temperature data obtained at harvesting sites. Between 1988 and 1996, 422 infections were reported; 45% were wound infections, 43% primary septicemia, 5% gastroenteritis, and 7% from undetermined exposure. Eighty-six percent of patients were male, and 96% with primary septicemia consumed raw oysters. Sixty-one percent with primary septicemia died; underlying liver disease was associated with fatal outcome. All trace-backs with complete information implicated oysters harvested in the Gulf of Mexico; 89% were harvested in water >22 degrees C, the mean annual temperature at the harvesting sites (P < .0001). Control measures should focus on the increased risk from oysters harvested from the Gulf of Mexico during warm months as well as education about host susceptibility factors.

Adolescent↗
PubMed1998

The contribution of subtle oocyte or sperm dysfunction affecting fertilization in endometriosis-associated or unexplained infertility: a controlled comparison with tubal infertility and use of donor spermatozoa.

This study aims to determine the relative contribution of oocyte and/or sperm dysfunction to the reduction of fertilization rates in vitro in cases of minor endometriosis and prolonged unexplained infertility. The results of in-vitro fertilization (IVF) treatment with ovarian stimulation have been compared between couples with the above conditions and women with tubal infertility (as control for oocyte function) and the use of donor spermatozoa (as control for sperm function). Fertilization and cleavage rates using husband's spermatozoa were significantly reduced in endometriosis couples (56%, n = 194, P < 0.001) and further significantly reduced in couples with unexplained infertility (52%, n = 327, P < 0.001) compared with tubal infertility (60%, n = 509). Using donor spermatozoa the rates were the same as using husband's spermatozoa in tubal infertility (61%, n = 27) or endometriosis (55%, n = 21) but significantly though only partly improved with unexplained infertility (57%, n = 60, P < 0.02). In unexplained infertility, a significantly increased proportion of couples experienced complete failure of fertilization and cleavage in a cycle (5-6% versus 2-3%). However, complete failure was not usually repetitive, and the affected couples did not account for the overall reduction in fertilization and cleavage rates, which remained significantly lower in the rest of the unexplained and endometriosis groups. Implantation and pregnancy rates appeared similar in all groups. The benefit of IVF treatment in cases of minor endometriosis and prolonged unexplained infertility is due to superabundance of oocytes obtained by stimulation. The reduction in natural fertility associated with endometriosis appears to be at least partly due to a reduced fertilizing ability of the oocyte. In unexplained infertility, there is distinct impairment due to otherwise unsuspected sperm dysfunction but probably also oocyte dysfunction.

Adult↗
PubMed1998

Production of active chimeric pediocin AcH in Escherichia coli in the absence of processing and secretion genes from the Pediococcus pap operon.

Minimum requirements have been determined for synthesis and secretion of the Pediococcus antimicrobial peptide, pediocin AcH, in Escherichia coli. The functional mature domain of pediocin AcH (Lys+1 to Cys+44) is targeted into the E. coli sec machinery and secreted to the periplasm in active form when fused in frame to the COOH terminus of the secretory protein maltose-binding protein (MBP). The PapC-PapD specialized secretion machinery is not required for secretion of the MBP-pediocin AcH chimeric protein, indicating that in Pediococcus, PapC and PapD probably are required for recognition and processing of the leader peptide rather than for translocation of the mature pediocin AcH domain across the cytoplasmic membrane. The chimeric protein displays bactericidal activity, suggesting that the NH2 terminus of pediocin AcH does not span the phospholipid bilayer in the membrane-interactive form of the molecule. However, the conserved Lys(+1)-Tyr-Tyr-Gly-Asn-Gly-Val(+7)-sequence at the NH2 terminus is important because deletion of this sequence abolishes activity. The secreted chimeric protein is released into the culture medium when expressed in a periplasmic leaky E. coli host. The MBP fusion-periplasmic leaky expression system should be generally advantageous for production and screening of the activity of bioactive peptides.

ATP-Binding Cassette Transporters↗
PubMed1998

Isolation and characterization of pediocin AcH chimeric protein mutants with altered bactericidal activity.

A collection of pediocin AcH amino acid substitution mutants was generated by PCR random mutagenesis of DNA encoding the bacteriocin. Mutants were isolated by cloning mutagenized DNA into an Escherichia coli malE plasmid that directs the secretion of maltose binding protein-pediocin AcH chimeric proteins and by screening transformant colonies for bactericidal activity against Lactobacillus plantarum NCDO955 (K. W. Miller, R. Schamber, Y. Chen, and B. Ray, 1998. Appl. Environ. Microbiol. 64:14-20, 1998). In all, 17 substitution mutants were isolated at 14 of the 44 amino acids of pediocin AcH. Seven mutants (N5K, C9R, C14S, C14Y, G37E, G37R, and C44W) were completely inactive against the pediocin AcH-sensitive strains L. plantarum NCDO955, Listeria innocua Lin11, Enterococcus faecalis M1, Pediococcus acidilactici LB42, and Leuconostoc mesenteroides Ly. A C24S substitution mutant constructed by other means also was inactive against these bacteria. Nine other mutants (K1N, W18R, I26T, M31T, A34D, N41K, H42L, K43N, and K43E) retained from <1% to approximately 60% of wild-type activity when assayed against L. innocua Lin11. One mutant, K11E, displayed approximately 2. 8-fold-higher activity against this indicator. About one half of the mutations mapped to amino acids that are conserved in the pediocin-like family of bacteriocins. All four cysteines were found to be required for activity, although only C9 and C14 are conserved among pediocin-like bacteriocins. Several basic amino acids as well as nonpolar amino acids located within the hydrophobic C-terminal region also were found to be important. The mutations are discussed in the context of structural models that have been proposed for the bacteriocin.

ATP-Binding Cassette Transporters↗
PubMed1998

HIV, hepatitis B and sexual practices in the street-recruited injecting drug users of Calcutta: risk perception versus observed risks.

Injecting drug users (IDUs) were recruited from the streets of Calcutta to obtain a baseline biological and behavioural data on risk practices. One-fifth of them (mostly using buprenorphine) tested positive for hepatitis B surface antigen (HBsAg); 4% were reactive to serologic test for syphilis (VDRL: Venereal Disease Research Laboratory). Condom use was insignificant while 74% reported sex with female sex workers and 15% of male IDUs also reported having sex with men. Although, sharing of injecting equipment ('works') was perceived as dangerous by the IDUs, majority of them (90/103) reportedly shared it; cleaning of works before sharing was a concern for intravenous but not for intramuscular drug injecting. Half of the IDUs reported suffering ever from abscess; a proportion (12%) of which had had superadded attack of maggots in it. They were also found to be infected with HIV (1%, 95% CI 0.028-5.97%) at a low prevalence that prompted subsequent launching of needle syringe exchange programme, establishment of cleaning norms before sharing of works, cleaning of injecting site on the body and condom promotion.

Abscess↗
PubMed1998

Interaction of hydrostatic pressure, time and temperature of pressurization and pediocin AcH on inactivation of foodborne bacteria.

High hydrostatic pressure, because it can kill microorganisms, is being investigated for potential use as a nonthermal food preservation method. The objective of this study was to determine the hydrostatic pressurization parameters, pressure, time, and temperature, and a bacteriocin that in combination would destroy 7 to 8 log cycles of pathogenic and spoilage bacterial populations. We suspended cells of Staphylococcus aureus, Listeria monocytogenes, Salmonella typhimurium, Escherichia coli O157:H7, Lactobacillus sake, Leuconostoc mesenteroides, Serratia liquefaciens, and Pseudomonas fluorescens in peptone solution and exposed them to the combination of treatments. The combined parameters used were hydrostatic pressure (138 to 345 MPa), time (5 to 15 min), temperature (25 to 50 degrees C), and pediocin AcH (3,000 AU/ml, final concentration). In general, cell death increased as the pressure, time, or temperature increased; however, the cells developed proportionately greater sensitivity as the pressure increased to 276 MPa and higher and the temperature increased above 35 degrees C. Pressurization for longer than 5 min, especially at lower pressure and temperature ranges, had very little added benefit. Among the four gram-negative species, E. coli O157:H7 was the most resistant to pressurization while among the four gram-positive species, L. sake and L. mesenteroides had greater resistance. The death rate at high pressure (345 MPa) and high temperature (50 degrees C) in combination followed first-order kinetics; at lower pressure and temperature combination it showed a late tailing effect. Estimated D value data indicated that even at 345 MPa and 50 degrees C an 8-log-cycle viability loss could not be achieved within 5 min for all eight species. However, when pediocin AcH was included during pressurization this loss was achieved.

Bacteria↗
PubMed1996

Cell-wall polysaccharides from the marine green alga Ulva "rigida" (Ulvales, Chlorophyta)--NMR analysis of ulvan oligosaccharides.

In order to obtain information on sugar sequences in the water soluble polysaccharides from Ulva "rigida", oligosaccharides were purified by anion exchange and gel permeation chromatography from the partial acid hydrolysate of the native or desulfated ulvan. The chemical structure and sugar sequence of these oligomers were determined by 1H and 13C NMR spectroscopy to be alpha-L-rhamnosyl (1-->4) D-xylose, beta-D-glucuronosyluronic acid (1-->2)-alpha-L-rhamnosyl (1-->4) D-xylose, beta-D-glucuronosyluronic acid (1-->4) L-rhamnose 3 sulfate, beta-D-glucuronosyluronic acid (1-->4) [beta-D-glucuronosyluronic acid (1-->2)] L-rhamnose and beta-D-glucuronosyluronic acid (1-->4) [beta-D-glucuronosyluronic acid (1-->2)] alpha-L-rhamnosyl (1-->4) D-xylose. The sugar linkages and particularly the branching and the sulfate position were in total agreement with previous chemical results.

Carbohydrate Conformation↗