Alpha‐phellandrene‐induced DNA damage and affect DNA repair protein expression in WEHI‐3 murine leukemia cells in vitro

Although there are few reports regarding α‐phellandrene (α‐PA), a natural compound from Schinus molle L. essential oil, there is no report to show that α‐PA induced DNA damage and affected DNA repair associated protein expression. Herein, we investigated the effects of α‐PA on DNA damage and repair associated protein expression in murine leukemia cells. Flow cytometric assay was used to measure the effects of α‐PA on total cell viability and the results indicated that α‐PA induced cell death. Comet assay and 4,6‐diamidino‐2‐phenylindole dihydrochloride staining were used for measuring DNA damage and condensation, respectively, and the results indicated that α‐PA induced DNA damage and condensation in a concentration‐dependent manner. DNA gel electrophoresis was used to examine the DNA damage and the results showed that α‐PA induced DNA damage in WEHI‐3 cells. Western blotting assay was used to measure the changes of DNA damage and repair associated protein expression and the results indicated that α‐PA increased p‐p53, p‐H2A.X, 14‐3‐3‐σ, and MDC1 protein expression but inhibited the protein of p53, MGMT, DNA‐PK, and BRCA‐1. © 2014 Wiley Periodicals, Inc. Environ Toxicol 30: 1322–1330, 2015.     

Trends in Medicare Part D Medication Therapy Management Eligibility Criteria

Background

To increase the enrollment rate of medication therapy management (MTM) programs in Medicare Part D plans, the US Centers for Medicare & Medicaid Services (CMS) lowered the allowable eligibility thresholds based on the number of chronic diseases and Part D drugs for Medicare Part D plans for 2010 and after. However, an increase in MTM enrollment rates has not been realized.

Objectives

To describe trends in MTM eligibility thresholds used by Medicare Part D plans and to identify patterns that may hinder enrollment in MTM programs.

Methods

This study analyzed data extracted from the Medicare Part D MTM Programs Fact Sheets (2008–2014). The annual percentages of utilizing each threshold value of the number of chronic diseases and Part D drugs, as well as other aspects of MTM enrollment practices, were analyzed among Medicare MTM programs that were established by Medicare Part D plans.

Results

For 2010 and after, increased proportions of Medicare Part D plans set their eligibility thresholds at the maximum numbers allowable. For example, in 2008, 48.7% of Medicare Part D plans (N = 347:712) opened MTM enrollment to Medicare beneficiaries with only 2 chronic disease states (specific diseases varied between plans), whereas the other half restricted enrollment to patients with a minimum of 3 to 5 chronic disease states. After 2010, only approximately 20% of plans opened their MTM enrollment to patients with 2 chronic disease states, with the remaining 80% restricting enrollment to patients with 3 or more chronic diseases.

Conclusion

The policy change by CMS for 2010 and after is associated with increased proportions of plans setting their MTM eligibility thresholds at the maximum numbers allowable. Changes to the eligibility thresholds by Medicare Part D plans might have acted as a barrier for increased MTM enrollment. Thus, CMS may need to identify alternative strategies to increase MTM enrollment in Medicare plans.     

Human seminal relaxin is a product of the same gene as human luteal relaxin.

Unlike that of other species, which have only one gene encoding relaxin, the human genome contains two nonallelic genes for relaxin, designated H1 and H2, which encode markedly different relaxin peptides. Whereas human relaxin gene H2 is selectively expressed in the ovary, no ovarian expression of gene H1 has been detected. Since relaxin is actively produced in the human male, it is possible to postulate divergent gene expression of relaxin in the male and female. We examined this question directly through the structural determination of human seminal relaxin and its comparison with the structure of human luteal relaxin. Partially purified relaxin, prepared from pooled human seminal plasma which had been delipidated by extraction with acid acetone and hexane, subjected to two cycles of HPLC and an additional purification step by ion-exchange chromatography, was further purified by immunoaffinity chromatography, using a monoclonal antibody to the H2 relaxin A chain which cross-reacts with synthetic H1 relaxin, followed by an additional HPLC step performed on a C4 reverse-phase column. The recovered, purified relaxin was then analyzed by N-terminal gas-phase sequencing and fast atom bombardment mass spectroscopy for determination of the amino acid sequence and molecular ions of the A and B chains, respectively. The results demonstrate that the structure of the predominant relaxin in human semen plasma is derived from the product of the H2 gene, consisting of a N-terminal pyroglutamic acid A-24 A chain and a mixture of B-26 and B-27 B chains. With the exception of degradation of the seminal relaxin B chain C-terminus, this structure is identical to the structure of human luteal relaxin. Therefore, both human seminal and luteal relaxin are products of the H2 gene.     

Prognostic significance of in vitro marrow culture growth pattern in untreated acute nonlymphocytic leukemia

The growth pattern of marrow cells in agar culture was studied in 90 adult patients with acute nonlymphocytic leukemia (ANLL) at diagnosis. We classified the abnormal growth patterns into 4 groups, A: no growth, B: decreased growth, C: excessive microcluster formation and D: excessive cluster growth with more than 20 colonies. There was a good correlation between growth pattern and FAB subtype. A predominance of group A growth was observed in M1, while group B growth was found in 50% of patients with M2 and M5. No relationships between the growth patterns and other clinical parameters were detected. Sixty-six patients were evaluable for treatment outcome. The growth pattern significantly correlated with complete remission rate. The remission rates were 52, 87, 80, and 25% for patients with group A, B, C and D growth, respectively. Analyses of remission duration and survival curves showed significant differences among the different growth patterns. Patients with D growth experienced a shorter remission duration and a lower survival rate than other groups. These results indicate that the in vitro culture growth pattern in untreated ANLL is of prognostic significance in predicting the response to therapy.     

Clinical and angiographic variables affecting the progression of coronary artery disease as determined by quantitative angiography

To assess by serial quantitative angiography, the significance of clinical and angiographic variables that affect the progression of coronary artery disease (CAD). Progression of disease by sequential angiography is unpredictable and the role of clinical risk factors controversial. Various intervention trials have demonstrated less progression and even regression in hyperlipidemic patients. Correlates of progression have included a younger age, unstable angina, and greater involvement of the coronary arteries, with few studies looking at angiographic features of individual lesions. Serial angiograms on 74 patients were analyzed by computer assisted quantitative angiography using absolute measurements. A total of 99 diseased segments were analyzed for progression defined as an absolute reduction of 20% in luminal cross-sectional area. A preliminary correlation coefficient was calculated for each of the clinical and angiographic variables to detect any association with progression, and the odds ratio determined.The presence of any of the clinical risk factors-diabetes, hypertension, serum cholesterol, smoking, and a family history of coronary disease could not predict progression. The use of beta blockers was three times less likely to be associated with progression (odds ratio 0.33). While the presence of distal disease was associated with progression of a more proximal lesion (odds ratio 2.4), eccentricity, branch point location, lesion length, calcification, thrombus, or the presence of collaterals did not influence progression of disease in an individual segment. In conclusion, the presence of any of the clinical risk factors could not predict progression of disease in an individual coronary segment as determined by serial quantitative angiography, and the use of beta blockers and the absence of coexistent distal disease was associated with less progression of disease in an individual coronary segment. This may be related to changes in wall stress, reduced platelet interactions, and the integrity and permeability of the vascular endothelium to lipids.