The pathologic hallmarks of Alzheimer's disease (AD) include senile plaque, neurofibrillary tangles (NFTs), synaptic loss, and neurodegeneration. Senile plaque and NFTs are formed by accumulation of amyloid-β (Aβ) and hyperphosphorylated tau, respectively. Progressive synaptic dysfunction and loss closely correlate with cognitive deficits in AD. Based on studies of the genes responsible for familial AD and temporal patterns of pathologic changes in AD brains, the Aβ accumulation is thought to be a primary event that influences other AD pathologies in the developmental cascade of AD. However, the details of Aβ effects on the other AD pathologies remain poorly understood. In this review, we provide an overview of the effects of Aβ in AD brains, especially focusing on synaptic dysfunction and microglia. We have recently found abnormal accumulation of a key molecule for actin assembly in NFTs of AD brains, and it was revealed that the accumulation requires not only tau pathology but also an Aβ burden in a study using transgenic mouse models of AD. Synaptic integrity is morphologically maintained by the precise regulation of actin assembly. Therefore, the results suggest the possibility that Aβ may promote NFT maturation and induce synaptic dysfunction through the disturbance of actin assembly. Thus Aβ seems to be a promoting factor in brain aging. On the other hand, we have studied microglial phagocytic ability for a compensatory pathologic reaction to Aβ accumulation. Further studies on the Aβ-dependent AD pathologies may contribute to determining novel mechanisms of AD development and new therapeutic targets in AD.
Excessive Ca2+ elevation resulting from activation of NMDA and other Ca2+ channels is thought to play a pivotal role in pathologic events following brain ischemia. The Ca2+ elevation directly triggers necrotic or apoptotic cell death through activation of Ca2+/calmodulin (CaM)-dependent enzymes, including calcineurin (CaN). CaN, a Ca2+/CaM-dependent serine/threonine protein phosphatase, partly mediates apoptosis associated with neuronal death. In a mouse middle cerebral artery occlusion (MCAO) model, calpain, a Ca2+-dependent cysteine protease, converted CaN to the constitutively active form of 48 kDa in vivo. The calpain-induced CaN activation mediated delayed neuronal death through translocation of nuclear factor of activated T-cells (NFAT) and FKHR, a forkhead box class O family member (FOXO) into neuronal nuclei after brain ischemia. The FKHR activation occurred through decreased Akt activity with concomitant dephosphorylation by constitutively active CaN. Thereafter, FKHR formed a complex with CaN and in turn translocated into nuclei after brain ischemia. After nuclear translocation of NFAT and FKHR, the transcription factors stimulated expression of Fas-ligand by binding to its promoter regions. Taken together, constitutively active CaN mediates delayed neuronal death through Fas-ligand expression via up regulation of both NFAT and FKHR transcriptional activity in brain ischemia.
Parkinson disease is one of the most common neurodegenerative disorders and is characterized by the selective loss of dopaminergic neurons in the substantia nigra. Although endogenous dopamine itself could serve as a vulnerability factor for dopaminergic neurons, the mechanism by which dopamine contributes to dopaminergic neuronal death remains unknown. In addition, although a decrease in proteasome activity was found in patients with sporadic Parkinson disease, the relationship between the ubiquitin-proteasome system and dopaminergic neuronal death remains to be elucidated. Here we provide an overview of the roles of endogenous dopamine and proteasome activity in dopaminergic cell death. Treatment of catecholaminergic PC12 cells with the herbicide paraquat, a potential risk factor for the development of Parkinson disease, induced an increase in dopamine content, and depletion of intracellular dopamine suppressed paraquat-induced cytotoxicity. Although glutathione, which scavenged dopamine oxidation intermediate, provided almost complete protection against dopamine-mediated toxicity, catalase provided only partial protection against cell death caused by dopamine. These data suggest that the generation of dopamine oxidation intermediate, rather than that of reactive oxygen species, plays a pivotal role in dopamine-induced toxicity. Moreover, treatment with paraquat induced a decrease in proteasome activity, and proteasome inhibition suppressed dopamine-mediated cytotoxicity. Suppression of proteasome activity stimulated the NF-E2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway, and elevated γ-glutamylcysteine synthetase mRNA and glutathione content. Furthermore, suppression of the paraquat-induced increase in gluthathione content exacerbated paraquat toxicity. These results suggest that the reduction of proteasome activity may be involved in cellular defense mechanisms against dopamine-mediated paraquat toxicity.
Knowledge and techniques involved in medical affairs have been steadily advancing. The lifelong learning programs supported by Gifu Pharmaceutical University are introduced. The 3 unique programs consist of (1) a lifelong learning program concerning recent medical topics provided by our university, (2) a reeducation program containing some lectures and practices concerning the most advanced knowledge and techniques on pharmacy and medicine, provided by co-organization of 3 public universities (Nagoya City University, University of Shizuoka, and Gifu Pharmaceutical University), and (3) an annual lifelong learning program promoted by Gifu Pharmaceutical University Pharmacy. Gifu Pharmaceutical University Pharmacy accommodates 100 prescriptions daily from hospitals. The annual lifelong learning programs held by our pharmacy have comprehensively provided practical knowledge and techniques on newly developed medicines, pharmaceutical care practice, pharmacotherapy, community pharmaceutics, and so on, for the last 10 years. Pharmacists should have full responsibility for pharmacotherapy as health care workers. The pharmacists should make a concerted effort to understand pharmacotherapy through pharmaceutical care practice by cooperation with community pharmacists, hospital pharmacists, and pharmaceutical associations. Our lifelong learning program has contributed to the improvement of pharmaceutical skills and communication among pharmacists, medical doctors, and other health care workers.
New-generation pharmacists who graduate from the 6-year pharmacy education program will come into being in Japan in 2 years' time. The new program regards technical skills and caring attitudes suitable for healthcare professionals as important, as well as expert knowledge. Pharmacists are expected to become more involved in pharmacotherapy and patient care to overcome rural physician shortage and achieve better outcomes in pharmacotherapy. Pharmacists themselves also want to contribute to improve pharmacotherapy and patient care. Pharmacists educated with the former 4-year education program, however, hardly had a chance to learn clinical pharmacy or pharmaceutical care when they were pharmacy students. They have so far studied clinical knowledge, skills, and attitudes by themselves mostly after graduation. Therefore most pharmacists have not received systematic education or training about clinical pharmacy. Pharmacy schools employ pharmacists and physicians as professors, and built practical rooms for pre-clinical training to study pharmacy practice in recent years. We should use those human resources and laboratory equipment in pharmacy schools to facilitate recurrent education for pharmacists. Internet-based real time remote lecture is also useful for pharmacists working far from pharmacy schools to attend a recurrent class. I propose an education system in which pharmacists who completed the recurrent education program teach students pharmacy practice in their worksites, and both pharmacists and students are developing their practical skills to a high degree together.
It is important for clinics and hospitals to cooperate in treating cancer patients in the community health. We are treating cancer patients in cooperation with five general hospitals in Shizuoka and about 100 clinics in the same community. In this system, it is required that pharmacists in the community should have knowledge about beneficial effects and adverse events of anticancer drugs as do hospital pharmacists, and furthermore they should have good communication with cancer patients. The expectation for pharmacists is great in community medicine especially in the treatment of cancer patients.
The Tokai Regional Alliance Center for Recurrent Education supported by three public universities in Tokai area executes a continuing education program for pharmacists. One of the major features of this program is that it can deliver lectures easily to pharmacists using a web conferencing system. This system enables pharmacists to attend lectures at several remote places. The program consists of two learning courses. One is a lecture series given by teachers belonging to the university or university facilities. Another course is a training series using the university equipment. This training course incorporates the contents of a new 6-year pharmacy education program. Moreover, this training course is intended to facilitate participation of pharmacists to the health care team. This continuing professional development for pharmacists will not only improve their pharmaceutical expertise, but also will provide the education necessary for pharmacists to perform well in actual medical practice. Through these efforts the continuing education of future pharmacists will be strengthened by new educational contents provided by pharmaceutical universities.
Mast cells originate from hematopoietic stem cells and undergo terminal differentiation in the tissues, in which they are ultimately resident. Heterogeneity of tissue mast cells is, therefore, one of the key concepts for a better understanding of immune modulation by mast cells. Since no appropriate culture model has been developed for tissue mature mast cells, it was difficult to investigate the tissue-specific functions of mast cells. We established a novel cutaneous mast cell model by modifying the previously reported co-culture system with fibroblastic cell line. This model shares many characteristics with cutaneous mast cells, such as staining properties, sensitivity to cationic secretagogues, and higher levels of granule histamine and proteases. We extracted the candidate genes that should regulate differentiation and functions of mast cells by analyses of the gene expression profiles during the co-culture period. We further investigated the functions of cluster of differentiation 44 (CD44), which is the primary receptor of hyaluronan in mast cells, since CD44 was up-regulated during the co-culture period. Fluorescence study revealed that mast cells expressing CD44 were bound to the extracellular matrix containing hyaluronan and lack of CD44 impaired proliferation of the co-cultured mast cells. In the CD44−/− mice, the number of cutaneous mast cells was significantly decreased. Reconstitution analyses with the mast cell deficient strain revealed that CD44 expressed in mast cells should be required in the proliferation in the cutaneous tissues. In the next phase of mast cell research, it might become increasingly important to focus on the heterogeneity of tissue mast cells.
Tissue-resident mast cells are derived from circulating committed progenitors, which are originated from pluripotential hematopoietic stem cells in bone marrow. These progenitors migrate into extravascular tissues, where they undergo differentiation and maturation into tissue-specific mature phenotypes. When activated by IgE/antigen, stem cell factor, neuropeptides, or other stimuli, mature mast cells release three classes of biologically active products, including pre-formed mediators stored in secretory granules, newly transcribed cytokines and chemokines, and de novo synthesized lipid mediators. Therefore, these cells have been implicated as major effector cells in acute and chronic inflammatory diseases. In recent years, it has become clear that lipid mediators including arachidonic acid metabolites (prostaglandins and leukotrienes) and lysophospholipid-derived products play crucial roles in mast cell-associated pathology. In this article, we will provide an overview of the roles of various lipid mediators in allergic diseases fueled by studies of their biosynthetic enzymes or receptors. In the latter part, we will make a particular focus on phospholipase A2 enzymes, which are placed at the bottleneck (rate-limiting) step of the lipid mediator-biosynthetic pathways.
Zinc (Zn) is an essential nutrient and its deficiency causes growth retardation, immunodeficiency, and neuronal degeneration. Zn homeostasis is tightly controlled by transporting through Zn transporters and by buffering via metallothioneins, all of which are involved in the intricate regulation of Zn concentration and distribution in individual cells. Research in understanding of these molecules has progressed with application of genetic techniques, which allow us to clarify the diverse role of Zn in vivo and in vitro. However, the precise roles and molecular mechanism(s) of Zn's function in allergic response have not been clarified. Mast cells are granulated cells that play a pivotal role in allergic reactions. The granules of mast cells contain various chemical mediators and inflammatory cytokines that are released upon FcεRI crosslinking. In this article, I will describe a role of Zn/Zn transporter in FcεRI-mediated mast cell degranulation and cytokine production. Furthermore, Zn acts as an intracellular signaling molecule, that is, a molecule whose intracellular status is altered in response to an extracellular stimulus in mast cell, and that is capable of transducing the extracellular stimulus into an intracellular signaling event, like Ca2+. I have proposed that there are two classes of Zn signaling: “Early” and “Late” Zn signaling. In this review, I discussed how Zn and its homeostasis affect biological events especially for mast cell-mediated allergy response.
Stilbenoids such as resveratrol (3,5,4'-trihydroxystilbene) have drawn much attention due to the diversity of structures and biological activities. These compounds are typically found as oligomers in a few plant families, such as Dipterocarpaceae, Vitaceae, Leguminosae, Cyperaceae, and Gnetaceae. The rich structural variation and multifunctional bioactivity make stilbenoid oligomers interesting targets for detailed phytochemical investigations. The oligomeric stilbenoids in Dipterocarpaceaeous plants have been my main focus of extensive structural investigation for the past decade. The tetramers of a resveratrol such as (−)-hopeaphenol, vaticanol B, and vaticanol C are widespread and present in large quantities in Dipterocarpaceaeous plants. These are of special interest due to the large number of stereoisomers resulting from many asymmetric carbons and the various frameworks when a resveratrol is homogeneously oligomerized. The structural variations in Vatica, Vateria, Upuna, Cotylelobium, Dipterocarpus, Shorea, and Hopea genera have been examined and about 120 new resveratrol oligomers isolated to date. A detailed structural determination based on comprehensive spectral study has solved the difficulties in elucidation caused by the complicated stereochemistry that comprises diastereomers, epimers, enantiomers, and rotamers. The isolates bear a structural variation of fused cyclic frameworks including heterocyclic and bicyclo ring systems, and have been developed as a chemical library for drug discovery and chemical biology probes for the first time.
The control of molecular properties using external stimuli is an attractive research area that offers the potential for regulation of various biological phenomena. This review summarizes the concept, design, syntheses and properties of nucleic acid switches in response to external stimuli, namely, “external-stimuli-responsive bridged nucleic acids monomer”. From 1H-NMR experiments, every external-stimuli-responsive bridged nucleic acid monomer was found to have an N-conformation, while an S-conformation was predominantly observed after exposure to a proper stimulus. Each bridged nucleic acid monomer was effectively introduced into oligodeoxynucleotides using an automated DNA synthesizer. Moreover, oligonucleotides modified with these bridged nucleic acid monomer were changed in their hybridization property and tolerance to enzymatic digestion in response to each stimulus. These results clearly showed that external-stimuli-responsive bridged nucleic acids monomers should work as a nucleic acid switch, and have the potential for regulation of various biological phenomena.
The pain experienced by cancer patients can be managed in 70-90% of cases by the World Health Organisation protocol for cancer pain. However, cancer pain treatment in Japan is not sufficiently effective. To use medicine safely and effectively, various problems must be solved. Therefore, in this study, appropriate usage of cancer pain treatment was examined. We were able to use acetaminophen suppositories (800 mg each) in cancer pain patients. It was suggested that high serum concentrations of oxycodone and hydrocotarnine might be observed in geriatric patients or in the state of decreased hepatic blood flow, making dose adjustment is necessary for such patients. We also clarified that the conversion ratio from oral oxycodone to intravenous ocycodone/hydrocotarnine was 0.71±0.12. In addition, we clarified the pharmacokinetics of controlled-release oxycodone in patients with cancer pain. Moreover, the findings of our study indicate that in the steady state, the serum concentrations of fentanyl are not maintained at a constant level for 3 days following the use of transdermal fentanyl. We established a method of appropriately passing a nasal duct for sustained release of fine granules of morphine sulfate. Resolution of the clinical problems associated with cancer pain treatments is anticipated to allow the proper use of cancer pain treatments in Japan.
This study conducted a thorough examination on the associations among several factors of asthma therapy, i.e., physicians' compliance with the guidelines, patients' adherence to medication, asthma symptom severity, asthma episodes, and patient background. Fifty outpatients treated continuously for asthma from October 2002 to October 2004 at Showa University Hospital were selected and their medical charts were surveyed. Physicians were recognized as noncompliant when their treatments included divergence from the Asthma Prevention and Management Guidelines. Patient adherence was evaluated as the ratio of the “measured (dispensed)” doses divided by the “expected (prescribed)” doses. The inhaled corticosteroids adherence and the ratio of the asthma-related emergency department visits of the patients with a family asthma medical history were significantly higher (p=0.034) and lower (p=0.043), respectively, than those without this medical history. This may indicate the necessity of education for patients with no family history of asthma. A significant correlation between the mean patients' adherence and the asthma symptom severity at the end of surveillance was found (p=0.010), suggesting the importance of patient adherence in asthma control. The asthma symptom severity in the noncompliant group at the beginning of surveillance was significantly worse (p=0.016), suggesting that physician compliance was low when the asthma symptom severity was poor. Based on the above findings, we proposed a flow-chart, which includes the confirmation processes of patients' adherence to medication and physicians' compliance to guidelines, in order to better control asthma, while also taking their family medical history into account.
The present study investigated risk factors and subjective symptoms associated with drug-induced leucopenia. We selected 248 patients with drug-induced leucopenia from the Case Reports of Adverse Drug Reactions and Poisoning Information System (CARPIS) database of over 47000 case reports of adverse drug reactions and assigned them to a case group. We also randomly selected 743 cases of adverse drug reactions not associated with leucopenia as a control group. A comparison of patient characteristic data between the two groups using logistic-regression analysis revealed that female sex, autoimmune disease and renal damage were background risk factors for drug-induced leucopenia. In addition, thiamazole, ritodrine, propylthiouracil, ticlopidine, allopurinol, minocycline and captopril administration significantly increased the risk of drug-induced leucopenia. A significant association was also found for fever, chills and pharyngeal abnormalities. Based on these findings, we developed two estimated regression equations to help prevent drug-induced leucopenia in the community pharmacy setting.
The aim of this study was to develop an effective learning system for patient interview training as a part of a pre-education clinical pharmacy. We devised a learning system for performing pharmaceutical care training and then investigated its usefulness. The learning content was a first interview with a simulated patient (SP). All students were divided into 8 groups of 5. Each student practiced interviewing an SP while 2 other students checked the performance of the interviewer, with roles rotated within each group. Additionally, the teachers also rotated among the groups to check the students. We evaluated the results contained in 55 check sheets used by teachers to evaluate the learning system, 223 check sheets used by students, and 110 check sheets used by the SPs. We found that there was a significantly greater number of students rated as unable to perform by the teachers as compared to those rated by the other students. In addition, the ratings for the items, “other symptom is confirmed” and “the severity and properties of the symptom are confirmed” were similar to the above result. Furthermore, there was a significantly greater number of students rated as unable to perform by the teachers as compared to those rated by the SPs in regard to the item “interviewed using open-ended questions.” After the students had performed their first attempt at a first interview, 28.6% were rated as unable to perform by the teachers, which was significantly reduced to 15.8% after the fourth attempt and 10.4% after the fifth attempt. Our results indicate that students must practice the first interview at least 4 times before reaching a level of competency. In addition, our findings suggest that both teachers and SPs should undertake pre-education training in clinical pharmacy practice, as the evaluations were significantly different among the teachers, students, and SPs in the present study.
Drugs are sometimes covered with oblate or agar jelly. It is said that the medicinal effect of drugs covered with oblate is slow, but no studies have reported results confirming this. Therefore, we examined the dissolution behavior when the drug was covered with oblate or agar jelly. Three types of commercially available formulations of benzodiazepine were used: medazepam sugarcoated tablets, prazepam uncoated tablets, and clorazepate dipotassium capsules. Dissolution tests were performed using solutions of pH 1.2 and 5.6 to simulate normal gastric juice and gastric anacidity, respectively. Drugs covered with oblate were tested by the paddle method, and those covered with agar jelly were tested using the rotating basket method. Dissolution of clorazepate capsules not covered with oblate increased by approximately 10% when the pH was adjusted from 1.2 to 5.6, while those of medazepam and prazepam tablets decreased by approximately 40-60%. In contrast, the dissolution decreased significantly at both pH values for each drug covered with oblate. Dissolution further decreased when the amount of oblate was doubled. No detectable dissolution of medazepam tablets or of clorazepate capsules occurred when the drug was covered with agar jelly. Dissolution of prazepam tablets covered with agar jelly was only about 10% at the end of the test. These results indicate that dissolution is slowed and prolonged when a drug is covered with oblate or agar jelly, permitting sustained release of the drug. But, it is necessary to improve a suitable method for the dissolution.