Inquiry into the ideal function of the pharmacy in home care*

Background: The aim of this study is to analyze why home‐care services provided by pharmacists have not been effectively utilized. Method: Questionnaires were submitted to home‐care service users, physicians, visiting nurses and home‐helpers and pharmacy directors. We studied whether gaps existed between users’ needs, physicians’ expectations of pharmacy services and pharmacists’ awareness of the importance of pharmacy services. We also investigated whether a failure to recognize the importance of cooperation with pharmacists in home‐care provision existed among physicians and nurses/home‐helpers. Results: Users and physicians expect pharmacists to be more involved in counseling about home care and welfare services than home‐visiting services. Pharmacists recognize home visiting services as being of greater importance than counseling about home care and welfare services. The results indicated that gaps existed between users’ needs, the physicians’ expectations and pharmacists’ awareness of the importance of pharmacy services. In terms of cooperation with pharmacists, study results implied that: (i) nurses/home‐helpers’ awareness of pharmacists’ home‐visiting service is lower than that of physicians; (ii) physicians’ expectations regarding pharmacists’ participation in home care services is lower than that of nurses/home‐helpers; (iii) over 70% of both groups recognize the necessity of pharmacists’ home‐visiting service. Conclusions: Pharmacists need to get more involved in counseling users about home care and welfare. Also, there should be a special focus on heightening nurses/home‐helpers’ awareness of pharmacists’ home‐visiting service and on raising physicians’ expectations for pharmacists’ participation in home care services to develop home‐care related pharmacy services in Japan.     

Evaluation of intravascular hemolysis with erythrocyte creatine in patients with cardiac valve prostheses

STUDY OBJECTIVES: To detect intravascular hemolysis in patients with cardiac valve prostheses. Erythrocyte creatine, a marker of erythrocyte age that increases with shortening erythrocyte survival, was evaluated with other hemolytic markers and hemodynamic parameters. DESIGN: Prospective study. Patients and measurements: Erythrocyte creatine was enzymatically assayed in 33 patients with prosthetic valves, including 15 patients with aortic valve replacement, 13 patients with mitral valve replacement, and 5 patients with double-valve (aortic and mitral) replacement, and 33 control subjects. Blood flow velocity and valvular regurgitation were determined by Doppler echocardiography. Other hemolytic markers (lactate dehydrogenase [LDH], reticulocyte count, and haptoglobin) and cardiac muscle markers (myoglobin and myosin light chain 1) were also measured. RESULTS: Erythrocyte creatine and LDH levels were significantly higher (p < 0.0001) and the haptoglobin level was lower (p < 0.0001) in patients with a prosthetic valve as compared with control subjects. However, there were no significant differences in these markers between those with (n = 17) and without (n = 16) regurgitation. Patients with high erythrocyte creatine levels (> 1.8 micro mol/g hemoglobin) exhibited significantly higher total peak flow velocity (sum of peak flow velocities at mitral and aortic valves) than those with normal erythrocyte creatine levels (p = 0.006). Erythrocyte creatine had a significant correlation with total peak flow velocity (r = 0.64, p < 0.0001), but LDH and haptoglobin had no significant correlation with total peak flow velocity. Patients with high LDH levels (> 460 IU/L) showed significantly higher myoglobin (p = 0.008) and myosin light chain 1 (p = 0.02) than those with normal LDH levels, whereas erythrocyte creatine was not related to cardiac muscle markers. CONCLUSIONS: Erythrocyte creatine is a quantitative and reliable marker for intravascular hemolysis in patients with prosthetic valves. Mild hemolysis is ascribable to valvular flow velocity rather than regurgitation.     

Transmitochondrial mice as models for primary prevention of diseases caused by mutation in the tRNALys gene

We generated transmitochondrial mice (mito-mice) that carry a mutation in the tRNA^Lys gene encoded by mtDNA for use in studies of its pathogenesis and transmission profiles. Because patients with mitochondrial diseases frequently carry mutations in the mitochondrial tRNA^Lys and tRNA^Leu(UUR) genes, we focused our efforts on identifying somatic mutations of these genes in mouse lung carcinoma P29 cells. Of the 43 clones of PCR products including the tRNA^Lys or tRNA^Leu(UUR) genes in mtDNA of P29 cells, one had a potentially pathogenic mutation (G7731A) in the tRNA^Lys gene. P29 subclones with predominant amounts of G7731A mtDNA expressed respiration defects, thus suggesting the pathogenicity of this mutation. We then transferred G7731A mtDNA into mouse ES cells and obtained F₀ chimeric mice. Mating these F₀ mice with C57BL/6J (B6) male mice resulted in the generation of F₁ mice with G7731A mtDNA, named “mito-mice-tRNA^Lys7731.” Maternal inheritance and random segregation of G7731A mtDNA occurred in subsequent generations. Mito-mice-tRNA^Lys7731 with high proportions of G7731A mtDNA exclusively expressed respiration defects and disease-related phenotypes and therefore are potential models for mitochondrial diseases due to mutations in the mitochondrial tRNA^Lys gene. Moreover, the proportion of mutated mtDNA varied markedly among the pups born to each dam, suggesting that selecting oocytes with high proportions of normal mtDNA from affected mothers with tRNA^Lys-based mitochondrial diseases may be effective as a primary prevention for obtaining unaffected children.Mitochondrial DNA (mtDNA) carrying a large-scale deletion (ΔmtDNA) and single-point mutations in the tRNA^Lys gene and in the tRNA^Leu(UUR) gene causes chronic progressive external ophthalmoplegia (CPEO); myoclonic epilepsy with ragged-red fibers (MERRF); and mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS), respectively—the three most prevalent mitochondrial diseases (1–3). However, there are slight differences among the three disease phenotypes, even though these pathogenic mtDNA mutations all induce mitochondrial respiration defects. Considering that mitochondrial respiratory function is controlled by both mitochondrial and nuclear genomes (1–3), this controversial issue can be clarified by generating transmitochondrial mice (mito-mice) that share the same nuclear genetic background but carry different pathogenic mtDNA mutations corresponding to the mutations found in the three prevalent mitochondrial diseases. However, no well-established, effective protocols are available for introducing mutagenized mtDNA into the mitochondria of mammalian cells.In our previous studies (4–8) we found mtDNAs carrying pathogenic mutations in mouse cell lines, transferred them into mouse female germ lines, and generated several types of mito-mice, including mito-mice-Δ (4, 5) which harbor ΔmtDNA and therefore are disease models for CPEO. However, mito-mice harboring mtDNA with pathogenic mutations in the tRNA^Lys and tRNA^Leu(UUR) genes—and therefore prospective disease models of MERRF and MELAS, respectively—have not previously been established owing to the unavailability of mouse cell lines with corresponding tRNA mutations in mtDNA.To complement the paucity of effective technologies required for introducing mutagenized mtDNA into mitochondria of living mouse cells, we developed an alternative strategy involving cloning and sequence analysis to detect small amounts of mtDNA with somatic mutations in the mitochondrial tRNA genes. Because pathogenic mutations responsible for mitochondrial diseases occur preferentially in the tRNA^Leu(UUR) and tRNA^Lys genes of humans (1–3), we sequenced 43 clones generated from PCR products carrying the mitochondrial tRNA^Leu(UUR) and tRNA^Lys genes of P29 mouse lung carcinoma cells (9). One of the 43 clones had a somatic G7731A mutation in tRNA^Lys, which enabled the generation of transmitochondrial mito-mice expressing respiration defects for their use as models for diseases caused by mutations in the mitochondrial tRNA^Lys gene.     

Long-Term Effects of Angiotensin II Receptor Blockade with Valsartan on Carotid Arterial Stiffness and Hemodynamic Alterations in Patients with Essential Hypertension

Increased arterial stiffness and intima media thickness (IMT) in the common carotid artery (CCA) are related to cardiovascular risk in essential hypertension. Angiotensin II plays an important role in structural and functional changes in the vasculature. In this study, we evaluated the long-term effect of the angiotensin II receptor blocker, valsartan, on IMT, arterial stiffness, and hemodynamics in the CCA in patients with essential hypertension. A prospective 24 month study of treatment with valsartan (80–160mg/day) was performed in 24 hypertensive patients. An ultrasound of the CCA was carried out to determine IMT, the cross-sectional distensibility coefficient (CSDC), the carotid arterial stiffness index β, and diastolic flow velocity to systolic flow velocity ratio (Vd/Vs). Treatment with valsartan for 24 months reduced systolic and diastolic blood pressure significantly. Compared to baseline, the decrease in pulse pressure was greater after 24 months treatment than after 12 months treatment. Valsartan did not influence IMT; however, after 24 months, it caused a significant increase in CSDC and a decrease in stiffness index β compared to baseline. These changes were not observed after 12 months of treatment. In addition, Vd/Vs, a sensitive marker of relative diastolic blood flow, increased after 24 months' treatment with valsartan. These results suggest that long-term treatment with valsartan improves vascular wall function and hemodynamics in patients with essential hypertension.     

Effect of butyric acid on lung-colonizing ability of cloned low-metastatic Lewis lung carcinoma cells

The lung-colonizing ability of low-metastatic Lewis lung carcinoma cells (P-29) was enhanced by their in vitro treatment with butyric acid and its sodium salt, sodium butyrate. Of the short chain fatty acids tested, butyric acid was the most effective in enhancing the lung-colonizing ability of P-29 cells; propionic acid and valeric acid were slightly effective, but acetic acid and caproic acid were ineffective. The enhancing effect of butyric acid on the lung-colonizing ability of P-29 cells was reversible, indicating that the result was the consequence of epigenetic alterations. Treatment of P-29 cells with butyric acid resulted in enhancement of secretion of plasminogen activator, cellular cathepsin B activity, and cellular adhesiveness. The phenotypes of cells treated with butyric acid were compared with those of cells treated with dimethyl sulfoxide, which was reported to enhance the lung-colonizing ability of P-29 cells. Significant differences were found in the phenotypes, especially that of cellular adhesiveness; that is, butyric acid enhanced mainly homotypic aggregation of the cells, while dimethyl sulfoxide enhanced mainly heterotypic adhesion, such as adhesion to monolayers of endothelial cells. In addition, butyric acid reversibly caused hyperacetylation of core histones in P-29 cells, while dimethyl sulfoxide did not.