Paul P. Cook

MCF-7 breast cancer cells transfected with protein kinase C-alpha exhibit altered expression of other protein kinase C isoforms and display a more aggressive neoplastic phenotype.

Increased protein kinase C (PKC) activity in malignant breast tissue and positive correlations between PKC activity and expression of a more aggressive phenotype in breast cancer cell lines suggest a role for this signal transduction pathway in the pathogenesis and/or progression of breast cancer. To examine the role of PKC in the progression of breast cancer, human MCF-7 breast cancer cells were transfected with PKC-alpha, and a group of heterogenous cells stably overexpressing PKC-alpha were isolated (MCF-7-PKC-alpha). MCF-7-PKC-alpha cells expressed fivefold higher levels of PKC-alpha as compared to parental or vector-transfected MCF-7 cells. MCF-7-PKC-alpha cells also displayed a substantial increase in endogenous expression of PKC-beta and decreases in expression of the novel delta- and eta-PKC isoforms. MCF-7-PKC-alpha cells displayed an enhanced proliferative rate, anchorage-independent growth, dramatic morphologic alterations including loss of an epithelioid appearance, and increased tumorigenicity in nude mice. MCF-7-PKC-alpha cells exhibited a significant reduction in estrogen receptor expression and decreases in estrogen-dependent gene expression. These findings suggest that the PKC pathway may modulate progression of breast cancer to a more aggressive neoplastic process.

Phorbol esters induce death in MCF-7 breast cancer cells with altered expression of protein kinase C isoforms. Role for p53-independent induction of gadd-45 in initiating death.

Protein kinase C (PKC) modulates growth, differentiation and apoptosis in a cell-specific fashion. Overexpression of PKC-alpha in MCF-7 breast cancer cells (MCF-7-PKC-alpha cell) leads to expression of a more transformed phenotype. The response of MCF-7 and MCF-7-PKC-alpha cells to phorbol esters (TPA) was examined. TPA-treated MCF-7 cells demonstrated a modest cytostatic response associated with a G1 arrest that was accompanied by Cip1 expression and retinoblastoma hypophosphorylation. While p53 was detected in MCF-7 cells, evidence for TPA-induced stimulation of p53 transcriptional activity was not evident. In contrast, TPA treatment induced death of MCF-7-PKC-alpha cells. Bryostatin 1, another PKC activator, exerted modest cytostatic effects on MCF-7 cells while producing a cytotoxic response at low doses in MCF-7-PKC-alpha cells that waned at higher concentrations. TPA-treated MCF-7-PKC-alpha cells accumulated in G2/M, did not express p53, displayed decreased Cip1 expression, and demonstrated a reduction in retinoblastoma hypophosphorylation. TPA-treated MCF-7-PKC-alpha cells expressed gadd-45 which occurred before the onset of apoptosis. Thus, alterations in the PKC pathway can modulate the decision of a breast cancer cell to undergo death or differentiation. In addition, these data show that PKC activation can induce expression of gadd45 in a p53-independent fashion.

The impact of penicillin skin testing on clinical practice and antimicrobial stewardship

BACKGROUND Penicillin skin testing (PST) is a simple and reliable way of diagnosing penicillin allergy. After being off the market for 4 years, penicilloyl-polylysine was reintroduced in 2009 as PRE-PEN. We describe the negative predictive value (NPV) of PST and the impact on antibiotic selection in a sample of hospitalized patients with a reported history of penicillin allergy. METHODS We introduced a quality improvement process at our 861-bed tertiary care hospital that used PST to guide antibiotic usage in patients with a history consistent with an immunoglobulin E (IgE)-mediated reaction to penicillin. Subjects with a negative PST were then transitioned to a β-lactam agent for the remainder of their therapy. NPV of skin testing was established at 24-hour follow-up. We are reporting the result of 146 patients tested between March 2012 and July 2012. RESULTS A total of 146 patients with a history of penicillin allergy and negative PST were treated with β-lactam antibiotics. Of these, only 1 subject experienced an allergic reaction to the PST. The remaining 145 patients tolerated a full course of β-lactam therapy without an allergic response, giving the PST a 100% NPV. We estimated that PST-guided antibiotic alteration for these patients resulted in an estimated annual savings of

Impact of regular collaboration between infectious diseases and critical care practitioners on antimicrobial utilization and patient outcome.

82,000. CONCLUSION Patients with a history of penicillin allergy who have a negative PST result are at a low risk of developing an immediate-type hypersensitivity reaction to β-lactam antibiotics. The increased use of PST may help improve antibiotic stewardship in the hospital setting.

Sustained reduction in antimicrobial use and decrease in methicillin-resistant Staphylococcus aureus and Clostridium difficile infections following implementation of an electronic medical record at a tertiary-care teaching hospital

Objective:Antimicrobial stewardship programs have been shown to help reduce the use of unnecessary antimicrobial agents in the hospital setting. To date, there has been very little data focusing on high-use areas, such as the medical ICU. A prospective intervention was done to assess guideline compliance, antimicrobial expenditure, and healthcare cost when an infectious disease fellow interacts regularly with the medical ICU team. Design:A 3-month retrospective chart review was followed by a 3-month prospective intervention the following year. Two hundred forty-six total charts were reviewed to assess generally accepted guideline compliance, demographics, and microbiologic results. Setting:Twenty-four-bed medical ICU at an 861-bed tertiary care, university teaching hospital in North Carolina. Subjects:Patients receiving antibiotics in the medical ICU. Intervention:During the intervention period, the infectious disease fellow reviewed the charts, including physician notes and microbiology data, and discussed antimicrobial use with the medical ICU team. Measurements and Main Results:Antimicrobial use, treatment duration, Acute Physiology and Chronic Health Evaluation II scores, length of stay, mechanical ventilation days, and mortality rates were compared during the two periods. Results:No baseline statistically significant differences in the two groups were noted (i.e., age, gender, race, or Acute Physiology and Chronic Healthcare Evaluation II scores). Indications for antibiotics included healthcare-associated (53%) and community-acquired pneumonias (17%). Significant reductions were seen in extended-spectrum penicillins (p = 0.0080), carbapenems (p = 0.0013), vancomycin (p = 0.0040), and metronidazole (p = 0.0004) following the intervention. Antimicrobial modification led to an increase in narrow-spectrum penicillins (p = 0.0322). The intervention group had a significantly lower rate of treatments that did not correspond to guidelines (p < 0.0001). There was a reduction in mechanical ventilation days (p = 0.0053), length of stay (p = 0.0188), and hospital mortality (p = 0.0367). The annual calculated healthcare savings was

Safety and Completion Rate of Short-Course Therapy for Treatment of Latent Tuberculosis Infection

89,944 in early antibiotic cessation alone. Conclusion:Active communication with an infectious disease practitioner can significantly reduce medical ICU antibiotic overuse by earlier modification or cessation of antibiotics without increasing mortality. This in turn can reduce healthcare costs, foster prodigious education, and strengthen relations between the subspecialties.

Long-term effects of an antimicrobial stewardship programme at a tertiary-care teaching hospital

OBJECTIVES we evaluated the effect of implementation of an electronic medical record (EMR) on the use of antimicrobial agents and on the rates of infections with Clostridium difficile and methicillin-resistant Staphylococcus aureus (MRSA). METHODS this was a retrospective, observational study conducted between 1 January 2005 and 31 December 2009. Antimicrobial drug use, rates of nosocomial C. difficile infection (CDI) and MRSA infection, the number of medical charts reviewed and number of antimicrobial recommendations made and accepted were compared before and after implementing the EMR utilizing interrupted time-series analysis. RESULTS compared with the 10 quarters prior to implementing the EMR, there was a 36.6% increase in the number of charts reviewed (P < 0.0001), a 98.1% increase in the number of antimicrobial recommendations made (P < 0.0001) and a 124% increase in the number of recommendations accepted (P < 0.0001). There was a 28.8% decrease in the use of 41 commonly used antibacterial agents (P < 0.0001). Nosocomial CDI decreased by 18.7% (P = 0.07) and nosocomial MRSA infections decreased by 45.2% (P < 0.0001) following implementation of the EMR. CONCLUSIONS adoption of an EMR facilitated a significant increase in chart reviews and antimicrobial recommendations, which resulted in a sustained decrease in antimicrobial use. There were decreased nosocomial infections with MRSA and a trend towards decreasing CDIs following implementation of the EMR.

Ceftaroline ‐ a cause for neutropenia

BACKGROUND Nine months of isoniazid therapy is the recommended regimen for treatment of latent tuberculosis infection, but low completion rates are a serious problem. The search for shorter regimens, compared with the standard isoniazid regimen, is of vital importance. We describe our experience using short-course regimens for the treatment of latent tuberculosis infection. METHODS We conducted a nonrandomized, observational study of 459 patients in a county health department from June 2000 to January 2006. Short-course therapy was defined as pyrazinamide and rifampin taken daily or twice weekly for 2 months or rifampin taken daily for 4-6 months. Conventional therapy consisted of a 9-month regimen of isoniazid. Liver function testing was performed for both groups in accordance with clinical guidelines. Treatment completion and hepatotoxicity (according to the World Health Organization classification) were determined for the short-course and conventional therapy groups. RESULTS Treatment was completed by 241 (77.7%) of 310 patients in the short-course group and by 98 (65.8%) of 149 patients in the isoniazid group (P = .009). Moderate to severe hepatotoxicity (grades 3 and 4) occurred in 6.1% of patients receiving short-course therapy and in 2.0% of patients receiving isoniazid (P=.09). The hepatotoxicity observed in the short-course group was confined to patients receiving pyrazinamide and rifampin daily and was self limited in all cases after the medications were discontinued. CONCLUSIONS The rate of treatment completion was significantly higher with short-course regimens, compared with the isoniazid regimen. Although the overall risk of hepatotoxicity in patients receiving pyrazinamide and rifampin daily for the treatment of latent tuberculosis infection was higher, liver functions returned to normal after the medications were discontinued.

Reduction in Fluoroquinolone Use following Introduction of Ertapenem into a Hospital Formulary Is Associated with Improvement in Susceptibility of Pseudomonas aeruginosa to Group 2 Carbapenems: a 10-Year Study

Antimicrobial stewardship has been shown to reduce unnecessary antibiotic use, but there are few data on the long-term benefits of such a programme. Antimicrobial use over a 13-year period since implementing an antimicrobial stewardship programme (ASP) at our institution was examined. Nosocomial rates of Clostridium difficile infection (CDI) and antimicrobial susceptibility patterns of common nosocomial micro-organisms over the same period were also reviewed. Total antimicrobial use decreased by 62.8% (P<0.0001). There were decreases in use of aminoglycosides (-91.3%; P<0.0001), cephalosporins (-68.3%; P<0.0001), extended-spectrum penicillins (-77.7%; P<0.0001), macrolides (-27.2%; P=0.002), clindamycin (-95.9%; P<0.0001) and quinolones (-78.7%; P<0.0001). Antifungal use decreased by 71.0% (P<0.0001). There were increases in the use of carbapenems (+736%, P<0.0001) and anti-MRSA drugs (+73.3%; P<0.0001). There was a 56.7% (P=0.007) reduction in nosocomial MRSA infections. Nosocomial CDI rates decreased by 42.6% (P=0.005) between 2003 and 2010 and then increased to near baseline levels following implementation of more sensitive testing for detection of CDI in 2011. There were decreases in the rate (-71.9%; P=0.001) and percentage (-51.4%; P<0.0001) of quinolone-resistant Pseudomonas aeruginosa. There were decreases in the rate (P<0.0001) and percentage (P=0.02) of carbapenem-resistant P. aeruginosa following implementation of a policy restricting ciprofloxacin use. We have demonstrated sustained reductions in both antimicrobial use and drug-resistant organisms following implementation of an ASP.

Resveratrol Inhibits KSHV Reactivation by Lowering the Levels of Cellular EGR-1

The US Food and Drug Administration approved ceftaroline in 2010 for the treatment of community‐acquired pneumonia and skin and soft‐tissue infections. The most common adverse reactions are diarrhoea, nausea and rash. To present the first case of neutropenia directly related to ceftaroline.