The relationship of TP53 R72P polymorphism to disease outcome and TP53 mutation in myelodysplastic syndromes.

Nonsynonymous TP53 exon 4 single-nucleotide polymorphism (SNP), R72P, is linked to cancer and mutagen susceptibility. R72P associations with specific cancer risk, particularly hematological malignancies, have been conflicting. Myelodysplastic syndrome (MDS) with chromosome 5q deletion is characterized by erythroid hypoplasia arising from lineage-specific p53 accumulation resulting from ribosomal insufficiency. We hypothesized that apoptotically diminished R72P C-allele may influence predisposition to del(5q) MDS. Bone marrow and blood DNA was sequenced from 705 MDS cases (333 del(5q), 372 non-del(5q)) and 157 controls. Genotype distribution did not significantly differ between del(5q) cases (12.6% CC, 38.1% CG, 49.2% GG), non-del(5q) cases (9.7% CC, 44.6% CG, 45.7% GG) and controls (7.6% CC, 37.6% CG, 54.8% GG) (P=0.13). Allele frequency did not differ between non-del(5q) and del(5q) cases (P=0.91) but trended towards increased C-allele frequency comparing non-del(5q) (P=0.08) and del(5q) (P=0.10) cases with controls. Median lenalidomide response duration increased proportionate to C-allele dosage in del(5q) patients (2.2 (CC), 1.3 (CG) and 0.89 years (GG)). Furthermore, C-allele homozygosity in del(5q) was associated with prolonged overall and progression-free survival and non-terminal interstitial deletions that excluded 5q34, whereas G-allele homozygozity was associated with inferior outcome and terminal deletions involving 5q34 (P=0.05). These findings comprise the largest MDS R72P SNP analysis.

Impact of a cleaner-burning cookstove intervention on blood pressure in Nicaraguan women.

Few studies have evaluated the cardiovascular-related effects of indoor biomass burning or the role of characteristics such as age and obesity status, in this relationship. We examined the impact of a cleaner-burning cookstove intervention on blood pressure among Nicaraguan women using an open fire at baseline; we also evaluated heterogeneity of the impact by subgroups of the population. We evaluated changes in systolic and diastolic blood pressure from baseline to post-intervention (range: 273-383 days) among 74 female cooks. We measured indoor fine particulate matter (PM(2.5); N = 25), indoor carbon monoxide (CO; N = 32), and personal CO (N = 30) concentrations. Large mean reductions in pollutant concentrations were observed for all pollutants; for example, indoor PM(2.5) was reduced 77% following the intervention. However, pollution distributions (baseline and post-intervention) were wide and overlapping. Although substantial reductions in blood pressure were not observed among the entire population, a 5.9 mmHg reduction [95% confidence interval (CI): -11.3, -0.4] in systolic blood pressure was observed among women aged 40 or more years and a 4.6 mmHg reduction (95% CI: -10.0, 0.8) was observed among obese women. Results from this study provide an indication that certain subgroups may be more likely to experience improvements in blood pressure following a cookstove intervention.

"Beam me up, scotty": designing the future of nursing professional development.

Experts in nursing education, such as Benner, Sutphen, Leonard, and Day (2010), as well as the Institute of Medicine report, The Future of Nursing (2011), agree that 21st-century nursing education must change. However, the focus is often on academic education rather than practice education. Today's nurses deserve exceptional practice education that is built on 21st-century learning principles, technologies, and methods as they pursue lifelong, behavior-forming learning. This requires four key shifts: (1) using synergistic, collaborative, learner-centered methods; (2) infusing technology into learning and teaching; (3) designing professional development rooted in instructional design best practices; and (4) performing comprehensive evaluations of professional development. This article explores the necessity for change and encourages nursing professional development specialists to boldly go where they have never gone before.

Effects of sodium lactate and acetic acid derivatives on the quality and sensory characteristics of hot-boned pork sausage patties.

Sodium lactate and acetic acid derivatives were evaluated for their effects on color retention, microbial growth, and sensory attributes of hot-boned pork sausage patties. Treatments included: (a) sodium lactate (L), (b) buffered vinegar (V), (c) sodium lactate and vinegar mixture (LV), (d) control with BHA/BHT (C), and (e) negative control (NC). Treatments L and LV decreased TPC at day 14 and day 16 when compared to control samples and reduced bacterial numbers up to 18 days. In addition, use of lactate and vinegar increased (P<0.05) acceptability and juiciness and reduced (P<0.05) off-flavor and rancidity when compared to control treatments at day 14. These results revealed that the L and LV sausage patties retained sensory acceptability and shelf-life quality from day 14 through day 17 as opposed to other treatments. Additionally, sausage patties with LV maintained redness and sensory quality throughout 17 days of shelf-life, as compared to other treatments that retained color and quality for 14 days.

Regulatory T cell expression of CLA or α(4)ß(7) and skin or gut acute GVHD outcomes.

Regulatory T cells (Tregs) are a suppressive subset of CD4(+) T lymphocytes implicated in the prevention of acute GVHD (aGVHD) after allo-SCT (ASCT). To determine whether increased frequency of Tregs with a skin-homing (cutaneous lymphocyte Ag, CLA(+)) or a gut-homing (α(4)ß(7)(+)) phenotype is associated with reduced risk of skin or gut aGVHD, respectively, we quantified circulating CLA(+) or α(4)ß(7)(+) on Tregs at the time of neutrophil engraftment in 43 patients undergoing ASCT. Increased CLA(+) Tregs at engraftment was associated with the prevention of skin aGVHD (2.6 vs 1.7%; P=0.038 (no skin aGVHD vs skin aGVHD)), and increased frequencies of CLA(+) and α(4)ß(7)(+) Tregs were negatively correlated with severity of skin aGVHD (odds ratio (OR), 0.67; 95% confidence interval (CI), 0.46-0.98; P=0.041) or gut aGVHD (OR, 0.93; 95% CI, 0.88-0.99; P=0.031), respectively. This initial report suggests that Treg tissue-homing subsets help to regulate organ-specific risk and severity of aGVHD after human ASCT. These results need to be validated in a larger, multicenter cohort.

T lymphocyte subset abnormalities in the blood and lung in pulmonary arterial hypertension.

RATIONALE: Mounting data suggest that immune cell abnormalities participate in the pathogenesis of pulmonary arterial hypertension (PAH). OBJECTIVE: To determine whether the T lymphocyte subset composition in the systemic circulation and peripheral lung is altered in PAH. METHODS: Flow cytometric analyses were performed to determine the phenotypic profile of peripheral blood lymphocytes in idiopathic PAH (IPAH) patients (n=18) and healthy controls (n=17). Immunocytochemical analyses of lymphocytes and T cell subsets were used to examine lung tissue from PAH patients (n=11) and controls (n=11). MEASUREMENTS AND MAIN RESULTS: IPAH patients have abnormal CD8+ T lymphocyte subsets, with a significant increase in CD45RA+ CCR7- peripheral cytotoxic effector-memory cells (p=0.02) and reduction of CD45RA+ CCR7+ naive CD8+ cells versus controls (p=0.001). Further, IPAH patients have a higher proportion of circulating regulatory T cells (T(reg)) and 4-fold increases in the number of CD3+ and CD8+ cells in the peripheral lung compared with controls (p<0.01). CONCLUSIONS: Alterations in circulating T cell subsets, particularly CD8+ T lymphocytes and CD4+ T(reg), in patients with PAH suggest that a dysfunctional immune system contributes to disease pathogenesis. A preponderance of CD3+ and CD8+ T lymphocytes in the peripheral lung of PAH patients supports this concept.

Sequence conservation and distribution of the fusobacterial immunosuppressive protein gene, fipA.

Fusobacterium nucleatum is a gram-negative anaerobe involved in various diseases, including periodontitis. Recently, other investigators isolated the F. nucleatum FDC 364 fusobacterial immunosuppressive protein (FIP). One subunit, FipA, impairs T-cell activation in vitro and shows homology with beta-ketothiolases. However, its distribution and variability among fusobacteria was not reported. Cloned fipA gene sequences from F. nucleatum ssp. polymorphum (ATCC 10953) and F. nucleatum ssp. nucleatum (ATCC 23726) shared 89 and 92% identity, respectively, with FDC 364 fipA, and 90 and 94% identity, respectively, with the FDC 364 FipA predicted amino acid sequence. Southern blot analyses of chromosomal DNA from fusobacterial strains, including F. nucleatum and other Fusobacterium species, were performed using partial fipA sequences as probes. The results indicate that fipA is highly conserved among the F. nucleatum strains examined and that fipA homologues are widely distributed among fusobacteria. A clear relationship between immune suppression, metabolism and the FipA protein remains to be determined.

Expression of Streptococcus mutans wall-associated protein A gene in Chinese hamster ovary cells: prospect for a dental caries DNA vaccine.

The Streptococcus mutans strain GS-5 wall-associated protein A (Wap-A) is a precursor to the extracellular antigen A (AgA), a recognized candidate dental caries vaccine. The full-length wapA gene (wapA-E) and a C-terminal truncated version (wapA-G) encoding the AgA were cloned into the mammalian expression vector pcDNA 3.1/V5/His-TOPO. The resulting constructs were propagated in the Escherichia coli Top10. To investigate the expression of the S. mutans genes in mammalian cells, the above constructs were used to transfect Chinese hamster ovary (CHO) cells in the presence of the cationic lipid pfx-8. Transient expression of the wapA-E and wapA-G genes was observed at 24 h post-transfection, as shown by Western immunoblot analysis using a rabbit antiserum to S. mutans cell wall. Immunochemical staining of the transfected CHO cells showed expression of WapA mainly in the cells and budding vesicles, whereas AgA was found mainly in the transfected cells and extracellular medium. The expression of S. mutans proteins in CHO cells, in either vesicles or soluble form, suggested an antibody response to the above DNA constructs. Work is under way to test the efficacy of these as DNA vaccines against S. mutans.

Factors predicting return to work following mild traumatic brain injury: a discriminant analysis.

Studies of mild traumatic brain injury (MTBI) suggest that most individuals recover rapidly and return to their everyday activities. However, a percentage of MTBI patients report persistent problems with cognitive, physical, and emotional symptoms. There is also evidence that some experience changes in occupational functioning following MTBI. The current study used a stepwise discriminant function analysis (DFA) to examine the role of injury severity variables, cognitive performance, and ratings of symptoms of TBI in predicting work status following MTBI. Subjects included 121 MTBI patients who were all active-duty military personnel. The stepwise DFA revealed that age and three cognitive variables (verbal memory, verbal fluency, and a speed test of planning and strategy) were predictive of work status 3-15 months following a documented MTBI, correctly classifying work status 68.8% of the time. A cross-validation DFA was conducted, with a 66.1% correct classification rate. These findings highlight the importance of cognitive impairments in identifying those at risk for occupational impairment following MTBI.

High-level expression of a truncated wall-associated protein A from the dental cariogenic Streptococcus mutans.

Streptococcus mutans plays a primary role in the formation of dental caries. Previously, in our laboratory, an S. mutans genomic library was prepared, and the wapA gene was cloned into the shuttle vector, pSA4/4B2. To generate overexpression of wapA and to facilitate efficient purification of the WapA protein for use as an immunogen, an expression vector with the strong tac promoter was used. In order to answer questions regarding the optimization of solubility and expression based on gene size or the hydrophobicity of the protein product, 12 truncated constructs of the wapA gene were prepared using PCR. The truncated products were subcloned into the pGEX-6P-1 glutathione S-transferase (GST) fusion vector and expressed in E. coli BL21. The fusion proteins were analyzed by SDS-PAGE and confirmed by analysis with anti-GST and anti-WapA antibodies. Our study suggests that abrogation of the wapA promoter is necessary for expression of this gene in this expression system. Deletion of the signal peptide and the hydrophobic C terminus of WapA increased expression compared with the full-length construct, and truncation at the protease cleavage site of the C-terminal region greatly increased the stability of the protein without a loss in reactivity with the anti-WapA antibody. Western immunoblot analysis with anti-WapA antiserum clearly showed that the majority of the epitopes of the GST-WapA fusions are located in the N-terminal region of WapA. The immunogenicity of the various WapA fusion products is being examined in mice and rats to further map the immunologically dominant regions of the protein. This method effectively increased the expression of WapA and should contribute to the further understanding of gene expression of E. coli, as well as aid in the characterization of this protein for future immunologic evaluation.

Isolation and identification of nontuberculous mycobacteria from foods as possible exposure sources.

A variety of foods collected from local supermarkets and produce stands were examined as possible sources of nontuberculous mycobacterial exposure. Food samples were combined with sterile ultrapure water and manually shaken. To remove large particles, the suspensions were filtered through a sterile strainer, centrifuged, and the supernatants were discarded. The food pellets were stored at -75 degrees C. The pellets were treated with either oxalic acid or sodium hydroxide-sodium citrate solutions to reduce contamination by nonmycobacterial organisms. Decontaminated pellets were cultured on both Middlebrook 7H10C agar and Middlebrook 7H10C agar with supplemental malachite green. Plates were observed for growth at 2 and 8 weeks. Isolates demonstrating acid-fastness were identified to species using polymerase chain reaction and restriction enzyme analysis. Nontuberculous mycobacteria (NTM) were recovered from 25 of 121 foods. Six different species of NTM were isolated, the most predominant being Mycobacterium avium.

Native plasmids of Fusobacterium nucleatum: characterization and use in development of genetic systems.

Three native plasmids of Fusobacterium nucleatum were characterized, including DNA sequence analysis of one plasmid, pFN1. A shuttle plasmid, pHS17, capable of transforming Escherichia coli and F. nucleatum ATCC 10953 was constructed with pFN1. pHS17 was stably maintained in the F. nucleatum transformants, and differences in the transformation efficiencies suggested the presence of a restriction-modification system in F. nucleatum.

PCR comparison of Mycobacterium avium isolates obtained from patients and foods.

Mycobacterium avium is a cause of disseminated disease in AIDS patients. A need for a better understanding of possible sources and routes of transmission of this organism has arisen. This study utilized a PCR typing method designed to amplify DNA segments located between the insertion sequences IS1245 and IS1311 to compare levels of relatedness of M. avium isolates found in patients and foods. Twenty-five of 121 food samples yielded 29 mycobacterial isolates, of which 12 were M. avium. Twelve food and 103 clinical M. avium isolates were tested. A clinical isolate was found to be identical to a food isolate, and close relationships were found between two patient isolates and two food isolates. Relatedness between food isolates and patient isolates suggests the possibility that food is a potential source of M. avium infection. This study demonstrates a rapid, inexpensive method for typing M. avium, possibly replacing pulsed-field gel electrophoresis.

The nature of ethical expertise.

Critics of clinical ethicists sometimes claim that if there were expertise in ethics, then there would have to be objective moral knowledge. They also assume that there would be only one kind of ethics expertise, and that it would be a kind of professional specialization. All three assumptions are mistaken.

Bacterial eradication by clinafloxacin, CI-990, and ciprofloxacin employing MBC test, in-vitro time-kill and in-vivo time-kill studies.

Bactericidal activities of clinafloxacin, CI-990 and ciprofloxacin were examined and compared for a small collection of gram-positive and gram-negative pathogens. In-vitro antimicrobial activities were measured by MIC, MBC and time-kill studies; in-vivo efficacy of each drug was assessed in an acute lethal mouse infection model that monitored median protective dose, eradicative dose and time-kill in blood. In general, for all test methods, activity of clinafloxacin exceeded that of CI-990, which in turn exceeded that of ciprofloxacin. Improvement in clinafloxacin and CI-990 activity over ciprofloxacin was more apparent against gram-positive cocci; differences were less noticeable against Escherichia coli and Pseudomonas aeruginosa. Clinafloxacin activity was notably high against Staphylococcus aureus and, against ciprofloxacin-resistant strains, this high activity was maintained in vitro and measurable activity was demonstrated in vivo.

In-vitro activity of clinafloxacin, trovafloxacin, and ciprofloxacin.

Clinafloxacin and trovafloxacin are two new fluoroquinolones for which few comparative data are available. When MICs of ciprofloxacin against Gram-positive and Gram-negative nosocomial species were compared, clinafloxacin was the most potent although trovafloxacin was also more active than ciprofloxacin against Staphylococcus aureus and enterococci. All three drugs were bactericidal. Clinafloxacin displayed the lowest frequency of resistance, approximating 10(-11). Development of resistance studies over 13-14 passages in the presence of drug revealed a 32-fold increase in MIC of clinafloxacin against S. aureus compared with 512- and 1024-fold for trovafloxacin and ciprofloxacin respectively, although the three drugs were comparable against Enterococcus faecalis and the Gram-negative bacilli.