Overconsumption of Ginkgo biloba seeds induces food poisoning characterized by tonic-clonic convulsions and vomiting. The primary toxic component, 4′-O-methylpyridoxine (MPN), was purified from the seeds in 1985. This review includes the following aspects of ginkgo seed poisoning: 1) toxicity related to the content of MPN and MPN glucoside in G. biloba seeds; 2) the effect of MPN on vitamin B6 analogs, including an increase in pyridoxal and pyridoxic acid and decrease in pyridoxal-5′-phosphate plasma concentrations; 3) case reports of ginkgo seed poisoning in Asia, North America, and Europe, and their effective treatment via vitamin B6 administration. Considering the increase in the use of G. biloba seeds, it is essential to raise global awareness of their potential toxicity.
Mitogen-activated protein kinase (MAPK) is a highly conserved serine/threonine kinase that regulates multiple cellular processes such as cell proliferation, differentiation, apoptosis, and inflammation. Rnc1 has been identified as a regulator of Pmk1 MAPK signaling, a homologue of extracellular signal-regulated kinase (ERK)-1 MAPK in mammals. Rnc1 encodes a K-homology (KH)-type RNA-binding protein (RBP). Previously, it was reported that Rnc1 acts as a negative regulator of Pmk1 MAPK signaling through the mRNA stabilization of Pmp1, the MAPK phosphatase for Pmk1 in our laboratory. We analyzed the spatial regulation of Rnc1 and discovered that Rnc1 is exported from the nucleus by the mRNA-export system. The nuclear export of Rnc1 is important for exerting its function to stabilize Pmp1 mRNA. Therefore, the spatial regulation of Rnc1 affects MAPK signaling activity. We also reported that Nrd1, an RRM-type RBP, plays a critical role in cytokinesis by binding to and stabilizing myosin mRNA. Notably, Rnc1 and Nrd1 localize to stress granules (SGs) in response to various environmental stresses. Moreover, SG formation is inhibited in the Nrd1 or Rnc1 deletion cells, whereas the overproduction of Nrd1 or Rnc1, as well as that of mammalian RBP TIA-1, induces granule formation. These data show that Nrd1 and Rnc1 regulate SG formation as a novel SG component. Alterations of SG formation are linked to neurodegenerative diseases and resistance to anti-cancer drugs, thus conferring remarkable clinical importance to SGs. This review discusses the spatial regulation of RBPs or SG formation as novel targets for drug discovery.
Although the transcriptional modulator interferon-related developmental regulator 1 (Ifrd1) has been identified as a transcriptional coactivator/repressor in various cells, including bone-resorbing osteoclasts, no attention has been paid to its role in bone-forming osteoblasts. Therefore, in this study we show that Ifrd1 is a critical mediator of both osteoblastogenesis and osteoclastogenesis through its expression in osteoblasts. Ifrd1 deficiency enhanced both osteoblast differentiation and maturation, and increased the expression of Runt-related transcription factor 2 and Osterix. A coculture experiment revealed that Ifrd1 deficient osteoblasts have higher osteoprotegerin (OPG) expression and less ability to support osteoclastogenesis. These findings suggest that Ifrd1 plays a pivotal role in bone homeostasis through its expression in osteoblasts, and represents a therapeutic target for bone disease.
Osteoporosis increases the risk of bone fractures (e.g., the femur), reduces a person's activities of daily living (ADL) and increases the likelihood of being bedridden. Therapeutic drugs for osteoporosis include oral bisphosphonates and intravenous receptor activator of nuclear factor-κB ligand (RANKL) antibodies, both of which suppress osteoclast activity, as well as the subcutaneously administered recombinant human parathyroid hormone (PTH), which activates osteoblasts. However, there is currently no oral osteogenesis-promoting drug. In the present study, we found a low-molecular-weight compound, KY-273, with osteogenesis promoting effects. KY-273 induced osteoblast differentiation in ST2 cells and in rat bone marrow-derived mesenchymal stem cells at a dose of 0.1 μM. On the other hand, KY-273 did not clearly exert differentiation effects in osteoclasts, chondrocytes, adipocytes, or myoblasts. In ovariectomized rats, KY-273 clearly increased serum bone alkaline phosphatase (ALP) by at a dose of 3 mg/kg for 8 weeks, and increased both the cortical bone volume and medullary volume of the diaphyseal and epiphyseal regions of femoral bone, but did not affect trabecular bone. Although alendronate (used to decrease bone loss) increased trabecular bone, it did not have any significant effects on cortical bone. PTH increased epiphysis cortical and trabecular bone volume, and reduced medullary volume. KY-273 also displayed good oral absorption in rats. In conclusion, KY-273 is a promising candidate for use as an oral anti-osteoporosis drug with osteogenesis promoting effects.
The bone turnover marker (BTM) measurement in osteoporosis treatment includes evaluation of bone metabolism status or evaluation of bone loss risk level, determination of fracture risk, and evaluation of drug treatment. Currently, by using the BTM, it has become possible to evaluate and select an effective treatment for osteoporosis. The BTM has become widely used as a clinical test item in actual clinical practice. Patients' low adherence to osteoporosis medication regimens increases the risk of vulnerable fractures and affects the cost effectiveness of therapeutics. A joint working group has been established, with International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) and International Osteoporosis Foundation (IOF) in a central role. The joint policy document of the joint working group is intended to increase the international application of BTM in clinical medicine, and to eliminate blood type I procollagen-N-propeptide (P1NP) and type I collagen cross-linked C-telopeptide (CTX) in observational research and intervention studies, in order to eliminate the inherent uncertainty of these measurements in clinical use. Current osteoporotic drug treatment has been proven to prevent bone fractures, but poor adherence to dosage regimens is an ongoing problem in clinical practice; various attempts have been made to improve adherence. Low adherence to an osteoporosis medication regimen increases the risk of fracture, and affects cost effectiveness. The BTM is an effective indicator in monitoring reactivity to osteoporosis drug therapy, and can be used to individually evaluate guidelines for treatment continuity and medication. In addition, providing BTM information to patients has reportedly improved their adherence to therapeutics, thereby potentially improving both the outcome and cost-effectiveness of osteoporosis drug therapy.
Fragility fractures associated with osteoporosis can have a major impact on a person's “activities of daily living” (ADL) and life span. Therefore, osteoporosis and its complications must be addressed by early screening and prompt intervention. A Fracture Risk Assessment Tool (FRAX) has been developed by the WHO. FRAX allows the calculation/estimation of absolute fracture risk over a 10-year period. The FRAX can be completed using a free application form, accessible online. The relationship between FRAX and the young adult mean (YAM) was investigated in order to calculate the cut-off value of FRAX for each age group. FRAX can be used by both physicians and other healthcare professionals to pre-screen patients for more detailed subsequent work-ups. Using FRAX in clinical practice has proven to be simple, inexpensive and useful.
The destruction of β cells of pancreatic islets results in a reduced level of insulin secretion, thus resulting in the onset of diabetes. Diabetes caused by such a decrease in insulin secretion has been reported to be associated with mitochondrial dysfunction. Because of this, mitochondrial therapy would be expected to be a useful and productive strategy for the treatment of this disease. We previously reported the development of a MITO-Porter, a liposome-based nanocarrier that permits macromolecular cargos to be delivered into mitochondria via membrane fusion. In this presentation, we present our current findings on the development of a mitochondrial nanocarrier system aimed at the development of a novel method for treating and preventing diabetes. The system includes “a nanocarrier system for nucleic acids targeted to pancreatic β cells”, and “an in vivo system for the delivery of nucleic acids targeting the pancreas”. In this presentation, we propose the use of a “mitochondrial nanocarrier system” as a novel method for the treatment and prevention of diabetes, and discuss the contribution of mitochondrial nanocarrier systems to innovative drug development.
Although many treatments for type 2 diabetes mellitus (T2DM) have been developed, the quality of life for people with T2DM still tends to be lower than in those without the disease. Thus, the development of new T2DM treatments and prevention methods is required. Genetic predisposition and environmental factors are understood to be involved in the onset and pathology of T2DM. Therefore, we have attempted to explore genes and foods with potential for use in the treatment and prevention of T2DM. LipoQuality, which describes the functional features of diverse lipid species, has recently been a focus of study in the pathology of metabolic diseases. Phospholipids, the major components of biological membranes, are known to change in composition during the development of obesity and diabetes. Therefore, for our research, we focused on genes that regulate the composition of phospholipids. We examined the effects of such genes on T2DM using an improved adenovirus vector that demonstrates safer, higher, and longer-term transgene expression than that of the conventional adenovirus vector. We also found that certain foods inhibit the progression of non-alcoholic fatty liver disease, which is related to T2DM. In this review, we introduce our research results, demonstrating how genes and food independently contribute to the mechanisms of T2DM pathology.
Adiponectin, the most abundant adipose tissue-derived adipocytokine, improves insulin sensitivity and has anti-inflammatory properties. Disulfide-bond A oxidoreductase-like protein (DsbA-L) is a key molecule in the multimerization of adiponectin (i.e., activation of adiponectin). In mice, liver-specific knockout of the Dsba-L gene impaired the mitochondrial function in hepatocytes and exacerbated the high-fat-diet-induced fatty liver. In addition, the DsbA-L mRNA level is negatively correlated with body mass index (BMI) in humans. We recently investigated the clinical impact of the DsbA-L gene on lifestyle-related diseases in Japanese subjects. We confirmed the influence of the common DsbA-L rs1917760 polymorphism on the multimerization of adiponectin, as well as the association of the polymorphism with the risk of obesity and non-alcoholic fatty liver disease, using prediction models based on a non-linear mixed effect model and/or structural equation models among elderly participants in a health screening program. We also observed a decreasing effect of DsbA-L polymorphism on the DsbA-L mRNA level in peripheral blood mononuclear cells, and an increasing effect of the polymorphism on the prevalence of excessive weight among schizophrenia patients at a high risk for obesity. These findings suggest that DsbA-L may be a key molecule associated with the development and progression of obesity and its related diseases. Therefore, genotyping the DsbA-L polymorphism and identifying patients at a high risk of developing obesity may help prevent obesity and its complications by facilitating targeted prevention and treatment programs for high-risk individuals.
Epstein-Barr virus (EBV), a human oncogenic virus, is a B cell-tropic herpesvirus and has the ability to immortalize normal B cells during latent infection. The Epstein-Barr nuclear antigen 1 (EBNA1) protein of EBV is expressed in the most EBV latently infected cells and binds to a specific viral genome region termed “oriP” (origin of plasmid replication) to maintain the stability of the approximately 170 kb double-stranded circular virus genomic DNA (episome) in cells. EBV elimination is thought to inhibit progression of EBV-associated malignancies, and the EBNA1-dependent mechanisms for EBV episome replication and maintenance are considered to be novel molecular targets for anti-EBV therapy. We have explored small-molecule compounds that can inhibit the binding between EBNA1 protein and oriP and found one pyrrole imidazole polyamide named DSE3 which can also inhibit EBV-mediated immortalization of normal B cells. These data suggested that an EBNA1-targeting strategy could be useful to combat EBV-associated malignancies.
Kaposi's sarcoma-associated herpesvirus (KSHV) is the causative agent of Kaposi's sarcoma (KS), primary effusion lymphoma (PEL), and Castleman's disease. While liposomal doxorubicin has been used as an effective treatment for KS patients, the cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) regimen used for PEL patients was reported to have 1-year survival rates of less than 40%. Moreover, the development of anti-KSHV drugs inhibiting viral replication has been delayed. KSHV establishes a lifelong infection in its host and alternates between a “latent infection” and “lytic infection” state. Latent infection is associated with maintenance of the viral genome and transformation of the infected cells. Lytic infection is the process of producing infectious virus. Elucidating the KSHV life cycle and viral replication machinery is essential for developing novel therapeutic approaches and identifying potential drug targets. To tackle these issues, we have been screening for anti-PEL compounds using PEL-derived cell lines and utilizing recombinant KSHV for functional analysis of KSHV coding genes. In particular, we have focused on the “viral pre-initiation complex” of KSHV and determined its molecular mechanism. The coding proteins conserved among β- and γ-herpesviruses form a complex, which has functional homology with the pre-initiation complex of host cells. The complex is indispensable for the expression of viral proteins composing virus particles. This review summarizes the pathogenesis and therapies of KSHV-associated malignancies. Furthermore, we introduce our recent data on KSHV ORF34, which contributes to viral late gene expression via the formation of the viral pre-initiation complex.
Persistent infection with oncogenic human papillomaviruses (HPVs) is necessary for the development of cervical cancer, although the accumulation of somatic mutations in the host genome is also required for the generation of invasive cervical cancer. Recent studies have demonstrated concomitant genetic changes in the HPV genome; however, their relevance to cervical carcinogenesis is poorly understood. Here we review our recent study investigating the within-host genetic diversity of HPV and its relationship with cervical cancer progression through deep sequencing analyses of viral whole-genome sequences in clinical specimens. Intriguingly, HPV genomes within individual clinical samples show an astonishingly high level of nucleotide variation across all histological grades of cervical lesions. Among the various substitution patterns, C-to-T and C-to-A substitutions are the predominant changes observed in the HPV genomes. Analysis of the trinucleotide context for substituted bases reveals that TpCpN (N is any nucleotide), which is a preferred target sequence for the cellular apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like (APOBEC) proteins, is the major target for C-to-T substitutions in the HPV genomes. These mutational signature analyses strongly imply that within-host HPV variations are mostly generated through APOBEC-mediated mutagenesis. Because the HPV oncogenes E6 and E7 harbor APOBEC-related mutations, we propose a potential role for APOBEC-mediated mutagenesis in cervical carcinogenesis.
The development of antiviral agents enables the control of chronic infectious diseases caused by infection with herpesviruses, human immunodeficiency virus, and hepatitis C virus. In contrast, antiviral treatment against hepatitis B virus (HBV) infection remains a significant area for improvement. One of the main barriers hampering the progress of HBV research has been a lack of cell culture systems efficiently reproducing the viral proliferation process. Recently, cell line-based HBV infection systems have been developed which are useful to analyze the mechanisms of HBV replication and to screen for new anti-HBV agents. In this article, we summarize the establishment of such cell models and the identification of small molecules that inhibit the HBV entry process and discuss their future potential as a novel class of anti-HBV agents.
Hepatitis C virus (HCV) infection is a major leading cause of chronic severe liver diseases such as cirrhosis and hepatocellular carcinoma. The recent direct-acting antivirals (DAAs) for the treatment of HCV infection offer very high cure rates, but DAAs are vulnerable to drug resistance because HCV is an RNA virus, which generally has very high mutation rates. DAA resistance-associated variants of HCV could reduce the effectiveness of DAAs in the future. Thus, the continuous development of new anti-HCV drugs against different target molecules is needed. We have been studying the host factors involved in HCV entry into cells. From those studies, we obtained novel candidates for host-targeting anti-HCV entry inhibitors, such as monoclonal antibodies against HCV receptors, which can be used together with DAAs. In this symposium review, we present and discuss our recent work on anti-HCV strategies targeting HCV entry steps.
To enable community pharmacists to provide empathic patient counseling, we developed and validated a training program based on the cognitive behavioral therapy approach (CBT-A) in our previous study. The major focus points of the re-structured training program were “inclusion of basic communication skills”, “exemplifying correspondence involving CBT-A using pre-recorded video(VTR)”, and “approach methods for cases where counter-evidence is unavailable”. The training program lasted for 4 h per day, for a total of 8 h. We also performed role-play scenarios on information gathering and medication guidance for simulated patients before and after training, and evaluated patient satisfaction with counseling, patient counseling alliance scores, and the degrees of the psychological distance between patients and pharmacists. Participants had high satisfaction with the discussion and role-play aspects of the training, as in our previous study. Participants also showed high satisfaction with “exemplifying correspondence involving CBT-A using VTR”. Counseling time was significantly longer when using CBT-A compared to ordinary information gathering and medication guidance, but patient satisfaction and patient counseling alliance scores were both higher, and the psychological distance between patient and pharmacist was lower. Accordingly, if patients cannot solve their own problems, even when pharmacists provide polite responses and expertise, patients can be guided in their problem solving using CBT-A. It suggested that using CBT-A could solve the problem of patients with anxiety due to problems that cannot be solved only via drug-centered approach.
To clarify the associated factors for negative response to sumatriptan nasal spray in patients with cluster headache, we investigated the involvement of clinical information, such as the characteristics of headaches, before commencing sumatriptan nasal spray treatment. There were 18 male patients and 4 female patients. A total of 17 responders and 5 non-responders to sumatriptan nasal spray participated in the present study. Three factors for negative response to sumatriptan nasal spray, “young age of onset”, “psychiatric disorder”, and “the headache is not in the orbit,” were found. Oxygen inhalation and/or subcutaneous injection were effective for nonresponsive cases. Therefore, these factors are considered to be useful for predicting therapy before applying sumatriptan nasal spray.
To clarify the associated factors for negative response to sumatriptan nasal spray (NS) in patients with cluster headache, we investigated the involvement of clinical information before commencing NS treatment. The efficacy of NS was 77.3% (n = 17) for these 22 patients. Three factors for negative response to NS, "young age of onset ", "psychiatric disorder", and "the headache is not in the orbit," were found. Therefore, these factors are considered to be useful for predicting therapy before applying NS.
Benzodiazepine receptor agonists (BZDRAs) have been associated with an increased risk of falls in the elderly. However, the association between the elimination half-life (t1/2) of BZDRAs and the difference between benzodiazepines (BZDs) and non-benzodiazepines (Z-drugs) has not been clarified. By conducting a meta-analysis of observational studies, we compared the risk of falls with respect to 1) short-acting BZDRAs (t1/2<12 h) vs. long-acting BZDRAs (t1/2≥12 h) and 2) BZDs vs. Z-drugs in elderly patients. Data were retrieved from MEDLINE, the Cochrane Library, and Igaku Chuo Zasshi. In total, 13 observational studies from 12 articles were included in our study (short-acting BZDRAs, n=12; long-acting BZDRAs, n=9; BZDs, n=13; Z-drugs, n=7). The risk of falls was significantly increased by the use of short-acting BZDRAs [Odds ratio (OR) (95% Confidence interval (CI)): 2.00 (1.46-2.73)], long-acting BZDRAs [OR (95%CI): 2.16 (1.61-2.89)], BZDs [OR (95%CI): 1.67 (1.31-2.13)], and Z-drugs [OR (95%CI): 2.42 (1.35-4.34)] compared to the risk in BZDRAs non-users. The increased risk of falls in elderly patients was similar in each group and unrelated to t1/2. This study suggested that all BZDRAs including Z-drugs should be avoided in elderly patients.
It is expected that drug systems using nanoparticles will improve the problem of poor water solubility and bioavailability of lipophilic drugs. However, it is difficult to prepare the formulations containing nanoparticles, and it is important to determine the concentration and kind of additives to prepare the formulations. We previously reported that a nano pulverizer NP-100 is possible to prepare drug nanoparticles for the 2-3 min, and the cellulose derivatives (metolose®, methylcellulose) is usefulness to prepare the nanoparticles by the mill method. In this study, we investigated the relationships of methylcellulose type and crushing efficiency in NP-100. First, we demonstrated the effect of viscosity in the various methylcellulose on the ibuprofen (IBU, lipophilic drug) particle size, and showed that the viscosity did not relate the crushing efficiency by the NP-100. Next, we measured the changes of cumulative size frequency curve in IBU particles by the combination of the NP-100 and 0.1-2.0% methylcellulose (SM-4, 400 and 4000). The appropriate addition reached IBU nanoparticles, although, the appropriate addition amount of methylcellulose was different in the SM-4 (0.5%), 400 (1.0%) and 4000 (1.2%). In addition, the IBU became meringue-like when subjected to the bead mill method in the less of methylcellulose, and excessive addition of methylcellulose increased the ratio of coarse particle. In conclusion, this results show that the appropriate addition amount of methylcellulose is different in the type of methylcellulose, and these changes of cumulative size frequency curve is useful as index to determine the concentration and type of additives in the nanoparticle production.
When undergoing 90Y-ibritumomab tiuxetan (90Y-IT) treatment, patients are discharged from hospital soon after initiation of treatment and followed up as outpatients. Thus it is important to apprise patients of the safety information regarding 90Y-IT treatment. However, studies investigating the safety of 90Y-IT in real-world clinical practice are lacking. We sought to investigate the adverse events arising from 90Y-IT administration to patients in our hospital. Patients who received 90Y-IT treatment at Hiroshima University Hospital from April 2010 to December 2014 were eligible for this study. The medical records of the patients were reviewed retrospectively. Eleven patients (median age, 65 years) were enrolled. Patients were classified into 3 groups according to the number of prior regimens: 1, 2-3, or >3, consisting of 5, 4, and 2 patients, respectively. The number of patients with induced grade 3 and 4 hematotoxicity, respectively, was 5 and 0 for leukocytopenia, 3 and 2 for neutropenia, and 3 and 2 for thrombocytopenia. The median nadir time was 37 d for leukocytopenia, 37 d for neutropenia, 36 d for thrombocytopenia, and 43 d for anemia. Patients with 2 or more prior regimens tended to experience grade 3 or 4 hematotoxicity more frequently than those with 1 prior regimen. In conclusion, we showed that hematotoxicity is a major adverse event of 90Y-IT treatment and that the nadir time is later than that with conventional anticancer agents. Medical staff, including pharmacists, should direct attention to the initial symptoms of hematotoxicity, especially in those patients who have received several prior regimens.
Nail tips are nail art materials that can be attached to the nail with adhesives. Recently, nail/finger injuries related to nail tips have been reported and one of the causes is considered to be the adhesives used for attaching nail tips. The components of nail adhesives are mostly cyanoacrylate, which is also used as an industrial instant adhesive. During curing, cyanoacrylate adhesives release formaldehyde through hydrolysis. When it is marketed as a nail adhesive, there is no regulation regarding its formaldehyde amount nor obligation to indicate its ingredients in Japan. Additionally, a biological safety test is not required for nail adhesives. Thus, because the safety of nail adhesives is inadequately confirmed, it is necessary to investigate their biological safety. Therefore, we purchased 5 commercially available nail adhesives and 1 medical adhesive and examined their formaldehyde content and cytotoxicity. We examined the cytotoxicity of the adhesives in V79 cells by a colony forming assay. In this test, 5 nail adhesives showed higher toxicity than 1 medical adhesive. Formaldehyde concentrations in the extract of adhesives were as follows: 17.5 to 24.2 μg/mL for nail adhesives and 7.4 μg/mL for medical adhesives. Cyanoacetate did not elicit cytotoxicity at the final concentration up to 1000 μM. However, formaldehyde showed cytotoxicity, with an IC50 of 79 μM (2.4 μg/mL). Taken together, the cytotoxicity of nail adhesives could be due to the formaldehyde generated by the hydrolysis of cyanoacrylate. It seems important that nail adhesives will be regulated by obligation and enhanced safety in the future.