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1.  Probing the Subcellular Localization of Hopanoid Lipids in Bacteria Using NanoSIMS 
PLoS ONE  2014;9(1):e84455.
The organization of lipids within biological membranes is poorly understood. Some studies have suggested lipids group into microdomains within cells, but the evidence remains controversial due to non-native imaging techniques. A recently developed NanoSIMS technique indicated that sphingolipids group into microdomains within membranes of human fibroblast cells. We extended this NanoSIMS approach to study the localization of hopanoid lipids in bacterial cells by developing a stable isotope labeling method to directly detect subcellular localization of specific lipids in bacteria with ca. 60 nm resolution. Because of the relatively small size of bacterial cells and the relative abundance of hopanoid lipids in membranes, we employed a primary 2H-label to maximize our limit of detection. This approach permitted the analysis of multiple stable isotope labels within the same sample, enabling visualization of subcellular lipid microdomains within different cell types using a secondary label to mark the growing end of the cell. Using this technique, we demonstrate subcellular localization of hopanoid lipids within alpha-proteobacterial and cyanobacterial cells. Further, we provide evidence of hopanoid lipid domains in between cells of the filamentous cyanobacterium Nostoc punctiforme. More broadly, our method provides a means to image lipid microdomains in a wide range of cell types and test hypotheses for their functions in membranes.
doi:10.1371/journal.pone.0084455
PMCID: PMC3883690  PMID: 24409299
2.  International expert panel on inflammatory breast cancer: consensus statement for standardized diagnosis and treatment 
Annals of Oncology  2010;22(3):515-523.
Background: Inflammatory breast cancer (IBC) represents the most aggressive presentation of breast cancer. Women diagnosed with IBC typically have a poorer prognosis compared with those diagnosed with non-IBC tumors. Recommendations and guidelines published to date on the diagnosis, management, and follow-up of women with breast cancer have focused primarily on non-IBC tumors. Establishing a minimum standard for clinical diagnosis and treatment of IBC is needed.
Methods: Recognizing IBC to be a distinct entity, a group of international experts met in December 2008 at the First International Conference on Inflammatory Breast Cancer to develop guidelines for the management of IBC.
Results: The panel of leading IBC experts formed a consensus on the minimum requirements to accurately diagnose IBC, supported by pathological confirmation. In addition, the panel emphasized a multimodality approach of systemic chemotherapy, surgery, and radiation therapy.
Conclusions: The goal of these guidelines, based on an expert consensus after careful review of published data, is to help the clinical diagnosis of this rare disease and to standardize management of IBC among treating physicians in both the academic and community settings.
doi:10.1093/annonc/mdq345
PMCID: PMC3105293  PMID: 20603440
guidelines; inflammatory breast cancer; management
3.  Prognostic impact of discordance between triple-receptor measurements in primary and recurrent breast cancer 
Annals of Oncology  2009;20(12):1953-1958.
Background: We evaluated discordance in expression measurements for estrogen receptor (ER), progesterone receptor (PR), and HER2 between primary and recurrent tumors in patients with recurrent breast cancer and its effect on prognosis.
Methods: A total of 789 patients with recurrent breast cancer were studied. ER, PR, and HER2 status were determined by immunohistochemistry (IHC) and/or FISH. Repeat markers for ER, PR, and HER2 were available in 28.9%, 27.6%, and 70.0%, respectively. Primary and recurrent tumors were classified as triple receptor-negative breast cancer (TNBC) or receptor-positive breast cancer (RPBC, i.e. expressing at least one receptor). Discordance was correlated with clinical/pathological parameters.
Results: Discordance for ER, PR, and HER2 was 18.4%, 40.3%, and 13.6%, respectively. Patients with concordant RPBC had significantly better post-recurrence survival (PRS) than discordant cases; patients with discordant receptor status had similarly unfavorable survival as patients with concordant TNBC. IHC scores for ER and PR showed weak concordance between primary and recurrent tumors. Concordance of HER2–FISH scores was higher.
Conclusions: Concordance of quantitative hormone receptor measurements between primary and recurrent tumors is modest consistent with suboptimal reproducibility of measurement methods, particularly for IHC. Discordant cases have poor survival probably due to inappropriate use of targeted therapies. However, biological change in clinical phenotype cannot be completely excluded.
doi:10.1093/annonc/mdp263
PMCID: PMC2791352  PMID: 19596702
concordance; HER2; hormone receptors; survival; testing
4.  Influence of fluctuations of plasma large neutral amino acids with normal diets on the clinical response to levodopa. 
Plasma large neutral amino acids (LNAAs) compete with levodopa for entry into the brain. Fluctuations in plasma LNAA concentrations could therefore contribute to variability in clinical response to levodopa. The hourly plasma levodopa, plasma LNAAs and clinical response were investigated in 11 fluctuating Parkinsonian patients on a regular hospital diet. The fluctuations in plasma levodopa were 2 to 3 times greater than the fluctuations of plasma LNAAs. The correlation between clinical response and plasma levodopa was substantially improved in only one patient by considering plasma LNAAs and calculating relative levodopa flux into brain. Although plasma LNAAs significantly increased during the day, the patients' clinical status did not uniformly deteriorate and mean afternoon clinical scores correlated better with mean plasma levodopa and levodopa flux than with mean plasma LNAAs. Minimum effective concentrations of levodopa for clinical response did not correlate with 9 am LNAA concentrations. It is concluded that in most patients, the relatively small variation in plasma LNAAs in comparison with the large variations in plasma levodopa indicates that fluctuations in LNAA are not an important contributor to the fluctuating response to levodopa.
PMCID: PMC1032296  PMID: 2738591
5.  Seroepidemiology of cholera in Gulf coastal Texas. 
Single serum samples from 559 volunteers from a Texas Gulf Coast area were examined for vibriocidal antibody to Vibrio cholerae O1 (biotype El Tor, serotype Inaba) by a microtiter method. Elevated levels of vibriocidal antibody were present in 14% of the subjects. Also, 6.8% of the subjects had elevated levels of antibody to the enterotoxin of V. cholerae O1 by the immunoglobulin G enzyme-linked immunosorbent assay. Recent infection, defined on the basis of elevations in both vibriocidal and antitoxin antibodies, had occurred in 1.3% of the subjects. When subjects who reported Brucella infection, travel to a cholera-endemic area, and/or cholera vaccination within a year of the study were removed from the analysis, a prevalence of recent infection of 0.89% was obtained. Significantly higher titers of vibriocidal antibody were found in those with exposure to seawater (fishermen, shrimpers, merchant marines, and dock workers) than in those without such exposure (P less than 0.005). Furthermore, titers of antitoxin antibody were significantly higher in those who consumed shellfish than in nonconsumers. Finally, titers of vibriocidal antibody were significantly higher in Vietnamese subjects than in non-Vietnamese subjects. The results of this study indicate that an endemic focus of infection with V. cholerae occurs in this area.
PMCID: PMC202726  PMID: 3415232
6.  In vitro growth inhibition of mastitis pathogens by bovine teat skin normal flora. 
One factor contributing to differences in the susceptibility of cows to mastitis may be differences in the teat skin normal flora, which could inhibit or enhance the growth of pathogenic bacteria. Using in vitro cross-streaking methods, we found that 25% of the isolates of teat normal flora of non-lactating heifers inhibited the growth of selected mastitis pathogens, but enhancers were not detected. Gram-positive pathogens were inhibited to a greater extent than Gram-negative pathogens. Inhibition was not a characteristic of specific genera or species of normal flora, but rather a property of certain variants within a species. This phenomenon of inhibition of mastitis pathogens in vitro by normal flora may be useful as an in vivo biological control method to reduce the incidence of mastitis.
PMCID: PMC1255269  PMID: 3552170
7.  Bacterial interactions in bovine respiratory and reproductive infections. 
Journal of Clinical Microbiology  1985;21(5):803-807.
The ability of the aerobic bacterial flora from the normal bovine respiratory and reproductive tracts to enhance or inhibit the growth of Pasteurella haemolytica, P. multocida, and Haemophilus somnus was tested in vitro. Six strains of each of these pathogens were cross streaked with each isolate of bovine normal flora. Flora which enhanced the growth of these pathogenic bacteria outnumbered inhibitors four to one. An intermediate number of isolates produced no effect on pathogen growth. Most enhancers were gram positive (Micrococcus, Staphylococcus, Corynebacterium, or Rhodococcus isolates), although several isolates of Moraxella and Actinobacter were also good enhancers. For H. somnus, there were proportionally more organisms which produced marked enhancement among the preputial flora than among the nasal flora, which may account for the greater number of genital carriers than nasal carriers. Bacillus isolates were the most significant inhibitors among the nasal flora, whereas no genus or species from the reproductive tract was noted to produce appreciable inhibition. It is proposed that changes in ratios of inhibitors to enhancers may determine, in part, whether a carrier state or disease occurs. Also, suggestions are made for in vitro use of this phenomenon for diagnostic tests.
Images
PMCID: PMC271786  PMID: 3998115
8.  Survival and detection of rotaviruses on environmental surfaces in day care centers. 
Previously, we demonstrated that children in day care centers commonly experience diarrhea due to rotavirus, giardia, and bacterial pathogens. Multiple agents frequently coexist, and the environment is heavily contaminated with enteric bacteria during outbreaks. A study of environmental surface contamination with rotavirus was performed during three non-outbreak periods. Of 25 samples collected from environmental surfaces and teachers hands at a day care center, 4 (16%) were positive for rotavirus antigen when a fluorescence assay was used. We also examined the survival of two animal viruses, rotavirus SA-11 and poliovirus type 1, and bacteriophage 12 on similar environmental surfaces in a laboratory. Poliovirus type 1 and bacteriophage f2 were more resistant to drying than rotavirus SA-11 and could be recovered after a 90-min exposure on a dry surface. Rotavirus SA-11 could be detected for 30 min. All three viruses survived longer when they were suspended in fecal material than when they were suspended in distilled water. These data suggest that several agents, including rotavirus, can remain viable on contaminated surfaces long enough to be transmitted to susceptible children. This finding helps explain why rotavirus shows a mode of spread like that of parasitic and bacterial agents within day care center settings.
PMCID: PMC239472  PMID: 6314896
9.  Antibodies to heat-labile Escherichia coli enterotoxin in Apaches in Whiteriver, Arizona. 
Infection and Immunity  1975;12(6):1475-1477.
Antitoxin titers to heat-labile Escherichia coli enterotoxin were measured in Apache children hospitalized with acute diarrhea and in Apaches of different age groups without diarrhea in Whiteriver, Ariz. The study suggests that in this locale, exposure to enterotocigenic E. coli is probably widespread and occurs early in life. Antitoxin titer rises after idarrheal disease associated with enterotocigenic E. coli infection, however, were not regulary found.
PMCID: PMC415459  PMID: 1107221
14.  The Conscientious Objector 
British Medical Journal  1898;2(1977):1590.
PMCID: PMC2434665

Results 1-17 (17)