Cardiac redox homeostasis is precisely regulated by reactive oxygen species (ROS) or electrophilic molecules that are formed by ROS reacting with intracellular substrates, and their eliminating systems. We have focused on the role of nitric oxide (NO) generated from inducible NO synthase (iNOS) that is continuously upregulated from early stage of heart failure, and revealed that iNOS-derived NO acts as a protective factor in the early stage of heart failure, whereas it contributes to induction of cardiac early senescence in later stages. The switching mechanism of NO-mediated signaling includes formation of endogenous NO-derived electrophilic byproducts such as 8-nitroguanosine 3′,5′-cyclic monophosphate (8-nitro-cGMP), which selectively targets an oncogenic small GTPase H-Ras at Cys-184, leading to cardiac cell senescence via covalent modification (S-guanylation) and activation of H-Ras. We also found that hydrogen sulfide-related reactive sulfur species (RSS) function as potent nucleophiles to eliminate electrophilic modification of H-Ras and suppress the onset of chronic heart failure after myocardial infarction. Our results strongly suggest a new concept of redox biology in which suppression of electrophilic irreversible modification of protein cysteine thiols by RSS may be a new therapeutic strategy of cardiovascular diseases.
Hydrogen sulfide (H2S), the third known gaseous transmitter following nitric oxide and carbon monoxide, is generated by multiple enzymes including cystathionine-γ-lyase (CSE) in vivo. We previously demonstrated that H2S activates Cav3.2 T-type Ca2+ channels expressed on sensory neurons, leading to hyperalgesia and facilitation of inflammation. Here, we describe the role of H2S in processing of colonic pain and inflammation. Intracolonic (i.col.) administration of NaHS, an H2S donor, to mice evoked colonic pain-like nociceptive behavior and referred hyperalgesia accompanied by phosphorylation of ERK in the superficial layers of spinal dorsal horn, a marker for excitation of nociceptive neurons. The pronociceptive effect of NaHS was abolished by inhibitors or knockdown of Cav3.2 and by an inhibitor of TRPA1, another target molecule of H2S. In rats with colitis induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS), on the other hand, repeated i.col. administration of NaHS prevented colonic ulcer and inflammatory symptoms, which were inhibited by ablation of capsaicin-sensitive sensory neurons or T-type Ca2+ channel inhibitor. NaHS, given i.col., caused phosphorylation of ERK in the spinal dorsal horn of rats with TNBS-induced colitis, but not of naïve rats. In TNBS-treated rats, Cav3.2 was upregulated in the dorsal root ganglia, while CSE was downregulated in the colon. Taken together, these findings suggest that inhibitors of the CSE/H2S/Cav3.2 or TRPA1 pathways might be useful for the treatment of colonic pain diseases such as irritable bowel syndrome, while H2S donors or Cav3.2 activators might be useful for the treatment of inflammatory bowel disease including Crohn's disease.
Parkinsonism is a neurological syndrome characterized by tremor, hypokinesia, rigidity, and postural instability. The neurodegenerative condition of Parkinson's disease (PD) is the most common cause of parkinsonism. PD is classified as sporadic PD and familial PD. Whereas idiopathic PD is caused by a number of complex factors, familial PD is a result of mutations in PD-associated genes. Unraveling the mechanisms surrounding familial PD will offer pivotal clues in understanding etiology of not only familial PD but also sporadic PD. We have demonstrated neuroprotective effects with particular focus on DJ-1. On the other hand, idiopathic basal ganglia calcification, also known as Fahr disease (FD) is another condition characterized by parkinsonism. In 2012, solute carrier family 20A2 (SLC20A2) was identified as the causative gene for familial FD. Our analysis of patient samples revealed a novel mutation in SLC20A2. Type-III sodium-dependent phosphate transporter 2 (PiT-2), the protein encoded by SLC20A2, plays an important role in phosphate homeostasis. However, PiT-2's role in the pathology of FD remains largely unclear. We have established induced pluripotent stem (iPS) cells from FD patients and are investigating their usefulness in drug development. Here, we present some of our latest research findings.
Osteocytes are the most abundant cells in bone. However, little attention has been paid to their role in bone remodeling. In this study, osteoclast differentiation was significantly enhanced by conditioned media derived from cultures of osteocytic MLO-Y4 cells that were cultured under hypoxic conditions. Using microarray analysis, we identified growth differentiation factor-15 (GDF15) as a pivotal factor secreted from osteocytes under hypoxia. Indeed, treatment with recombinant GDF15 markedly increased osteoclast differentiation in vitro. Further to investigate the importance of GDF15 in vivo, we used a hypoxic murine model that involved ligation of the right femoral artery. The volume of cancellous bone in the proximal tibia of the ligated limb was significantly reduced, together with a significant increase in osteoclast-related parameters. Addition of anti-GDF15 antibody prevented bone loss and osteoclastic activation in the tibiae of mice that had undergone femoral artery ligation. These results suggest that GDF15, which is secreted from osteocytes under hypoxia during bone remodeling, may be a positive regulator of osteoclastic differentiation. The in vivo usefulness of the anti-GDF15 antibody might provide insights for the development of novel therapeutics for bone disorders related to hypoxia or ischemic insults.
This review concerns the taxonomic status of the genus Murraya in tribe Clausenae, subfamily Aurantioideae, family Rutaceae, and presents a new system integrating both morphology-based taxonomy and chemotaxonomy. This genus has been morphologically divided into the sects Murraya and Bergera. This dichotomy is justified by the noticeable difference of secondary metabolites with 3-prenylindoles in Murraya and carbazoles in Bergera. As for other metabolites of genus Murraya, coumarins are found in both sects, but differ clearly in types; 8-prenylcoumarins occur throughout the sect Murraya whereas geranylated furocoumarins are known from some species of the sect Bergera. As far as chemical properties are concerned, sect Bergera is much closer to genus Clausena than sect Murraya, suggesting the dichotomy of genus Murraya to be generic rather than sectional. 8-Prenylcoumarins characterizing sect Murraya play a decisive role in the distinction of M. exotica from M. paniculata that occurs most widely in subtropical and tropical Asia and is well known for morphologic as well as chemical diversity. Though the morphological difference between the two species is slight only in leaves and leaflets, the distinction is well substantiated by the following chemical feature: 7-OMe-8-prenylcoumarins occur in M. exotica whereas 5,7-di-OMe-8-prenylcoumarins in M. paniculata. Sect Murraya has a very close relation to genus Merrillia that is chemically characterized by similar types of 8-prenylcoumarins, and is also related to a certain extent to genus Micromelum. M. exotica is philologically surveyed in view of the delicate relationships between Okinawa, the only habitat of this plant in Japan, and China in order to clarify its historical background.
Hypervalent organo-λ3-iodanes have attracted great interest in organic synthesis; however, the chemistry of hypervalent organo-λ3-bromanes remains largely unexplored, mainly because of synthetic difficulties. Iodobenzene is readily oxidized by common oxidants such as peracids under mild conditions, whereas bromobenzene is inert. Since the ionization potential of bromobenzene (8.98 eV) is higher than that of iodobenzene (8.69 eV), hypervalent-λ3-bromane would show much higher reactivity. We overcame this problem by using difluoro(aryl)-λ3-bromane (Frohn's reagent), which serves as a pivotal progenitor of various λ3-bromanes including imino-λ3-bromane, cyclopent-1-enyl-λ3-bromane, (E)-β-alkylvinyl-λ3-bromane, and diacetoxy-λ3-bromane. These reagents possess highly unusual reactivity based on the enormously enhanced nucleofugality of the aryl-λ3-bromanyl group.
Morphine has been widely used for the treatment of acute, chronic, and cancer pain and is considered the strongest analgesic in clinical care. Conversely, morphine-induced analgesia may be accompanied by several side effects. Animal studies have demonstrated that low doses of morphine administered intrathecally can produce reliable analgesia for thermal, mechanical, and chemical nociceptive stimulation. On the other hand, high doses of morphine administered intrathecally may induce spontaneous nociceptive responses such as scratching, biting, and licking in mice as well as agitation and vocalization in rats. In addition, similar nociceptive responses including hyperalgesia, allodynia, and myoclonus have been observed in humans following intrathecal or systemic administration of high-dose morphine. It has been suggested that the spontaneous nociceptive behaviors evoked by high-dose morphine may be mediated by a non-opioid mechanism that is not yet fully understood. This review describes the mechanisms of spontaneous nociceptive behaviors evoked by high-dose morphine focusing on the neurotransmitters/neuromodulators released from primary afferent fibers.
Phenols are abundant in nature and diverse phenols are readily available commercially at low cost. Thus, phenols can be used as the raw materials for the synthesis of valuable multisubstituted aromatic compounds by the direct activation of phenolic hydroxyl groups (C-O bond activation), followed by substitutions with other substituents. Although the derivatization of phenolic hydroxyl groups to sulfonates, such as triflates, nonaflates, tosylates and mesylates, followed by the transition-metal-catalyzed coupling reactions has been extensively investigated for this purpose, the direct C-O bond activation of phenols for subsequent functional group transformation has been a long-standing challenge in modern organic synthesis. In this review, I have summarized my recent studies on the formal direct C-O bond activation of phenols using nonafluorobutanesulfonyl fluoride (NfF) for the synthesis of multisubstituted aromatics. I have focused on the dual use of NfF, a less expensive commercially available reagent, including the tentative formation of highly reactive nonaflates from phenols and the use of the liberated fluoride ion as a nucleophile to promote the reactions of nonaflates. The following four topics are discussed: 1) palladium-catalyzed coupling reactions of phenols, 2) novel preparation of benzynes from 2-silylphenols, 3) synthesis of fluorinated aromatic compounds via the formation of benzynes, and 4) Hiyama coupling of (tert-butyldimethylsilyl)arenes activated by internal phenolic hydroxyl groups.
Artificial nucleic acids have recently been widely used with their properties optimized for various technologies such as the inhibition of gene expression (antisense/antigene strategies, RNA interference) and genetic diagnosis (single nucleotide polymorphism (SNP), damaged nucleobase). For practical application of nucleic acid therapeutics, establishment of an effective delivery system for oligonucleotides is also required because of their poor permeability into cells. Various useful delivery technologies including lipoplexes formed using cationic lipids and polyplexes made with cationic polymers have been developed; however, there is no crucial tool for oligonucleotide therapeutics at present. If technologies of functional nucleic acids and adequate delivery systems are cooperatively developed, the realization of nucleic acid therapeutics might be effectively accelerated. Based on this concept, we have been cooperatively developing these technologies based on organic synthetic chemistry during the past decade. This paper summarizes our recent results: 1) development of a specific fluorescent probe for 8-oxoguanine; 2) synthesis and evaluation of a prodrug-type small interfering RNA (siRNA) molecule; and 3) targeted intracellular delivery of oligonucleotides via conjugation with receptor-targeted ligands.
We evaluated the effectiveness of small group discussion (SGD) in association with a drug abuse prevention program for junior high school students. The students first received a lecture about drug abuse prevention, then participated in SGD. The discussion focused on how to take action when tempted to abuse drugs. We gave a questionnaire 3 times; before and after the lecture (before SGD), and after SGD. Seventy-seven students replied to these questionnaires. After the lecture, knowledge about drug abuse was improved and all students answered that they had never abused drugs. However, in answer to a different question, a few students noted that they might use drugs in some situations. We consider it necessary to give more consideration to this problem. After the lecture, 35.5% of the students felt that they had definitely acquired skills for drug abuse prevention, whereas after the SGD this was increased to 73.7%. In addition, more than 75% of the students answered that the SGD program was useful since the opinions of other students could be heard. These results suggest that more students acquired skills to prevent drug abuse by participation in SGD. Our findings showed that SGD was useful and that the students were able to more effectively understand important concepts related to drug abuse prevention.
Encouraging self-medication is expected to reduce healthcare costs. To assess the current situation of self-medication practices in the general population, we conducted a questionnaire survey regarding the use of over-the-counter (OTC) medications or dietary supplements in 1008 participants (37% men; mean age, 64±13 years) from Ohasama, a rural Japanese community. A total of 519 (52%) participants used OTC medications or dietary supplements, with common cold medication (36%) and supplements (28%) such as shark cartilage products representing the most common choices. Stepwise logistic regression showed female gender, a higher frequency of visits from a household medicine kit distributor, dyslipidemia, and lower home systolic blood pressure levels as predictors for the use of such materials (chi-square values: 25.3, 12.6, 7.0, and 4.6, respectively; all p<0.03). Stratifying the participants according to the use of antihypertensive treatment showed a negative association between systolic blood pressure and the use of OTC medications or supplements only in participants being treated for hypertension. These results suggest that although the adoption rate of self-medication in Japan can be increased in rural areas, it may remain lower in urban areas. The present study clarifies the factors associated with the use of OTC medications or dietary supplements and indicates that appropriate self-medication practices might improve the control of hypertension, particularly in patients undergoing antihypertensive treatment.
Pharmaceutical education is suffering from the decentralization of students due to the establishment of new pharmaceutical universities, the shift to a six-year program from a four-year program in the colleges of pharmacy in 2006, and a decrease in the number of students. Combined, these have lead to academic failings, even at high-ranking universities. However, abundant knowledge and ability are necessary to pass the national examination for pharmacists. At Kobe Pharmaceutical University, a Basic Education Center for Pharmacy was instituted in 2006 for the purpose of supporting the scholastic abilities of students with various challenges, as well as for students in general. One approach at the Basic Education Center for Pharmacy, has been to offer supplementary lessons, and to provide additional opportunities for learning for the repeater. We offer supplementary lessons as an “Office Hours Class” and also use DVD-learning for remedial teaching. “Office Hours Class” is conducted in a question/answer format with a small number of students. In order to develop the DVD-learning system (pharmaceutical educational digital learning; PEDL), which promotes self-learning, our supplementary lectures and “Office Hours Class” lectures are recorded and edited on DVD media. The learning effect of using these systems, as determined by regular examination, shows that these have been helpful. As a result, we concluded that these supplementary learning programs are useful as a learning method to help students acquire necessary knowledge as potential pharmaceutical professionals, and to increase student motivation.
When teicoplanin (TEIC) is injected at the maintenance dose, a long period is required for achievement of the target plasma trough concentration because of its long elimination half-life. An initial loading dose is necessary for rapid achievement of an effective plasma trough concentration. Thus, we proposed that it is necessary for a pharmacist determine the initial loading dose of TEIC to reach an effective plasma trough concentration rapidly after its administration to a patient. In the present study, we evaluated the effectiveness of initial loading dose determination by pharmacists and physicians by comparing the achievement rate of target plasma trough concentrations (>15 μg/mL) and expression of adverse effects. Among 61 patients, 34 were treated according to an initial loading dose determined by a pharmacist (pharmacist intervention) and 27 were treated according to the treating physician's discretion (non-pharmacist intervention). The achievement rate of target concentrations was 91.2% (plasma trough concentration 23.3±5.3 μg/mL) in the pharmacist intervention group and 25.9% (plasma trough concentration 14.0±5.9 μg/mL) in the non-pharmacist intervention group. There was no difference in the incidence of adverse effects between the two groups. Also, we found that systemic inflammatory response syndrome (SIRS) may have a correlation with plasma trough concentrations of TEIC. We suggest that the SIRS score could become a means way of determining initial loading dose. These findings suggest that it is potentially effective for a pharmacist to determine this initial dose in order to rapidly achieve the target plasma trough concentration of TEIC.