Aspiration pneumonia remains a major cause of death in the elderly. However, fundamental and effective treatment has not been established yet. Onset of aspiration pneumonia is based on the presence of dysphagia, such as delayed triggering of the swallowing reflex. The swallowing reflex in the elderly is temperature sensitive, even if it is impaired. Swallowing reflex was delayed when the temperature of the food was close to body temperature. The actual swallowing time shortened when the temperature difference increases. The improvement of swallowing reflex by temperature stimuli could be mediated by the temperature-sensitive TRP channel. Administration of the TRPV1 agonists improves the delay of the swallowing reflex. Red wine polyphenols have been suggested to improve the swallowing reflex by increasing TRPV1 response. Food with menthol, an agonist of TRPM8 which is a cold temperature receptor, also decreased the delay in swallowing reflex. Olfactory stimuli, such as black pepper, can be a useful tool to improve swallowing reflex in people with lower ADL and consciousness levels. By combining these various sensory stimuli, we developed a protocol to start oral intake in patients with aspiration pneumonia This protocol shall continue to contribute to the ingestion of many older people.
Free fatty acids (FFAs) are not only essential nutrient components, but they also function as signaling molecules in various physiological processes. In the progression of genomic analysis, many orphan G-protein coupled receptors (GPCRs) are found. Recently, GPCRs deorphanizing strategy successfully identified multiple receptors for FFAs. In these FFA receptors (FFARs), GPR40 (FFAR1) and GPR120 are activated by medium- to long- chain FFAs. GPR40 is expressed mainly in pancreatic β-cell and mediates insulin secretion, whereas GPR120 is expressed abundantly in the intestine and regulates the secretion of cholecystokinin (CCK) and glucagons-like peptide-1 (GLP-1), it promotes insulin secretion. Due to these biological activity, GPR40 and GPR120 are potential drug target for type 2 diabetes and selective ligands have been developed. In this review, we provide recent development in the field and discuss their physiological roles and their potential as drug targets.
Gut lumen is continually exposed to many agents, including noxious compounds. The intestinal epithelia form a barrier between the internal and luminal (external) environments. Chemical receptors that detect the luminal environment are thought to play an important role as sensors and as modulators of epithelial cell functions. The Molecular analysis of various epithelial cell membrane receptor proteins has elucidated the sensory role of these cells in the gut chemosensing system. Nutrient sensing systems by these receptors in the small intestinal epithelia are thought to influence nutrient metabolism and local physiological function. Much less is known, however, about the physiological roles of chemosensing in the large intestine. We have investigated the contractile and secretory effects of short-chain fatty acids (SCFAs), the primary products of commensal bacteria, and the expression of SCFA receptors in the large intestine. The findings indicate that the epithelia in the large intestine also detect and respond to luminal contents, particularly bacterial metabolites, for host defense. We recently reported that luminal bitter tastants and odorants affect transepithelial ion transport in human and rat colon, and that putative receptors are expressed in colonic mucosa. In this review, we describe the secretory effects of chemical stimuli on lumen associated with the expression pattern of sensory receptors, focusing on the large intestine.
Recent evidence indicates that free amino acids are nutrients as well as acting as chemical transmitters within the gastrointestinal tract. Gut glutamate research is the most advanced among 20 amino acids. Free glutamate carries the umami taste sensation on the tongue and a visceral sensation in the gut, especially the stomach. In the field of taste physiology, the physiological meaning of the glutamate-derived chemical sense, the umami taste, has been proposed to be a marker of protein intake. Experimental evidence in gut glutamate physiology strongly supports this hypothesis. Free glutamate is sensed by the abdominal vagus and regulates gastrointestinal functions such as secretion and emptying to accelerate protein digestion. Clinical application of glutamate has also just begun to treat gastrointestinal disorders such as dyspepsia, ulcer, dry mouth and functional dyspepsia. In this review, we introduce recent advances in gut glutamate research and consider the possible contribution of glutamate to health.
Glutamate is known as the umami substance in the diet and umami taste has been traditionally preferred in East Asian countries. Recent our and others' studies showed that glutamate has potential to protect the gastrointestinal mucosa against noxious agents. In contrast, Helicobacter pylori infection and non-steroidal anti-inflammatory drugs (NSAIDs) are recognized as the two major causes of gastrointestinal diseases characterized by gastritis or gastrointestinal ulcers. We examined whether dietary supplementation of glutamate prevents the Helicobacter pylori infection- and NSAIDs-induced gastrointestinal damages in animal models. In this paper, we first review how these noxious agents develop gastrointestinal damages, and secondly discuss the possible candidates of protective factors as well as the mechanisms how glutamate prevents these gastrointestinal damages. We propose that our daily intake of glutamate has important roles in protecting the gastrointestinal mucosa against Helicobacter pylori and NSAIDs and possibly contributes to the maintenance of our healthy lives.
Adjuvant is originated from the Latin word “adjuvare” which means “help” in English to enhance the immunological responses when given together with antigens. The beginning of adjuvant was mineral oil which enhanced the immune response when it was given with inactivated Salmonella typhimurium. Aluminium salt was used to precipitate diphtheria toxoid and increased level of antibody response was demonstrated when administered with alum-precipitated antigens. Since 1930, aluminium salt has been used as DTaP (diphtheria-tetanus-acellular pertussis vaccine) adjuvant. Many candidates were tested for adjuvant activity but only aluminum salt is allowed to use for human vaccines. New adjuvant MF59, oil-in-water emulsion type, was developed for influenza vaccine for elderly (Fluad) and series of AS adjuvant are used for hepatitis B, pandemic flue, and human papiloma virus vaccines. Oil-adjuvanted influenza pandemic vaccines induced higher antibody response than alum-adjuvanted vaccine with higher incidence of adverse events, especially for local reactions. Alum-adjuvanted whole virion inactivated H5N1 vaccine was developed in Japan, and it induced relatively well immune responses in adults. When it applied for children, febrile reaction was noted in approximately 60% of the subjects, with higher antibodies. Recent investigation on innate immunity demonstrates that adjuvant activity is initiated from the stimulation on innate immunity and/or inflammasome, resulting in cytokine induction and antigen uptake by monocytes and macrophages. The probable reason for high incidence of febrile reaction should be investigated to develop a safe and effective influenza vaccine.
MF59-adjuvanted seasonal trivalent inactivated (ATIV) vaccine licensed since 1997 and MF59-adjuvanted pandemic H1N1 vaccines have been distributed to approximately 80M persons. Addition of the emulsion adjuvant to inactivated vaccine formulations provides for higher levels of antibody to the viral hemagglutinin (HA) in less responsive older adults, infants and children which, in the case of the pandemic vaccine, allowed only 3.75 μg of the HA to be immunogenic. The adjuvant also stimulates production of more broadly-reactive antibodies against strains that are mismatched to those in the vaccine, a potential advantage in the face of perennial influenza virus antigenic drift. In a field trial, ATIV was 89% efficacious in preventing laboratory-confirmed influenza in 6-<72 month old children, 81% more efficacious than the unadjuvanted control split vaccine while, in older adults, ATIV reduced community-acquired pneumonia and influenza hospitalizations in adults >65 years old by 23% compared to unadjuvanted vaccine, in an observational study. The effectiveness of MF59 adjuvanted split pandemic H1N1 vaccine was 74% overall. Unadjuvanted pandemic vaccine was poorly immunogenic in HIV-infected persons, whereas their responses to MF59-adjuvanted vaccine were similar to those of healthy controls. Analyses of the clinical trials and pharmacovigilance databases and observational studies have shown that while MF59-adjuvanted influenza vaccines are more locally reactogenic, they have not been associated with an increased risk for various adverse effects (AE) of special interest, including unsolicited neurological or autoimmune events.
In the 1990s, drug companies focused their resources on chemistry-based proprietary blockbuster compounds (small molecules) for chronic diseases that could bring in several billion dollars in a short period of time. Since then, the focus has turned to biologics (proteins/high MW molecules) such as anticancer agents, antibodies, and so on. Vaccines, in contrast, are a rather slow-growing market, administered only a few times per patient, low priced, and often undifferentiated. Due to the influenza scares of recent years, the above view has changed remarkably. According to some analysts, the annual growth of the current $2.2bn vaccine market will become almost 10 percent over the next 5 years. In 2009, Pfizer (US), in an effort to boost their small vaccine-related business, purchased Wyeth (US). In October 2010, Johnson & Johnson announced they were buying Crucell (Germany), the only vaccine maker who had remained independent. GSK (UK) holds the top spot in the vaccine market with a 25% share. Pfizer (US), Merck (US), Novartis (Switzerland), and Sanofi-Aventis (France) are next, while Johnson & Johnson has moved into the 6th position by purchasing Crucell. There is of course an essential therapeutic need for vaccines, however, why are major pharmaceutical companies now investing a significant amount of resources in the vaccine business? Vaccine development may take more time than that of small molecules, but they are less risky from an intellectual property standpoint, and complicated manufacturing processes create a high barrier to follow-on biologics/biosimilars. Also in Japan, since the recent influenza scares, there has been acceleration in movement and cooperation among industry and government, including lawmakers.
Today the number of international cycling races has increased markedly compared with the past, and inevitably there are many riders traveling all over the world to participate in these events. It is inevitable that riders often become exposed to many more drugs at the cycling venues or through the internet where a wide selection of drugs is available. According to Union Cycliste Internationale (UCI) regulations, riders must report to a Doping Control Officer (DCO) any medications taken in the 72 h preceding a race. High level international teams are very familiar with these regulations and submit these reports with little problem. Lower level teams with less expertise, however, tend to have much more trouble submitting these reports. The close cooperation with pharmacists is believed to promoting the healthy development of the sport. To improve anti-doping controls, I strongly believe that good communication among staff during the events to exchange information smoothly and also good education through regular seminars and symposia for the staff involved are definitely needed. In other words, we have to develop human resources to build up a firm foundation to constantly generate future leaders.
I am a team doctor of three competition groups including professional cycling team for ten years. The most troublesome issue as a sports doctor is the problem about doping. I cope thanks to a mobile telephone and an e-mail regardless of place and time, but introduce some examples because I still experience many doping “near miss” cases. In addition, there are problems in road competition spots as follows; 1) There are few team doctors. I am pressed by the consultation from plural teams, 2) An unexperienced doctor of the doping knowledge often prescribes prohibited drugs, 3) There are problems with no understanding of the medicine made in foreign countries, Chinese medicine, a generic drug, and supplements which obtained on the internet. I hope that anti-doping education in faculty of pharmaceutical sciences is made mandatory, sports pharmacists taken to the sports spot along increase, and a system and a database to teach local doctors and players quickly will be achieved in future.
After the Ministry of Edcation, Culture, Sports, Science and Technology of Japan has ratified the UNESCO “International Convention Fight against Doping in Sport” in December 2006, the government increased its support to Anti-Doping activities. About 5 years ago, the total number of doping control samples a year in Japan was around 2000, and this number was not enough to demonstrate Japanese athletes' cleanness to the rest of the world. However, after the government's ratification of the UNESCO international convention, the government increased its support both financially and politically. By receiving the increased support from the government, testing number has increased and reached 5000 samples a year. 5 years ago, our target athletes range was only international level athletes who compete in the Olympics or international events. As we expanded our testing numbers, the target range of the athletes was also expanded and national level athletes also became our targets. As a result, athletes without having adequate knowledge about anti-doping regulations became our target. This situation caused inadvertent anti-doping rule violation cases. Most of those anti-doping rule violations were the result of taking over-the-counter medicines, etc. In order to cope with those inadvertent anti-doping rule violation problems, we, Japan Anti-Doping Agency launched “Sport Pharmacist Project” in cooperation with Japan Pharmaceutical Association. In this project, we provide anti-doping information/regulation to the pharmacists and make those pharmacists knowledgeable about the current anti-doping rules and regulations.
Recently, the sports pharmacist system was established and, a great deal of attention has been paid by students. However, the learning opportunity of anti doping education at the university is still rare. There is some experience of dope test staff's chaperon in an anti doping activity as a student. Through the experience of the chaperon, we can learn about a chain of flows of the doping inspection, characteristics of the sports games, and the desire of the player who cannot learn by the seat studies. I will describe the impression of the experience of the chaperon, the meaning, utility, and the view of the future.
Drug use and abuse by athletes has become a common problem. Pharmacists can assist by managing the legitimate medication needs of athletes to prevent them from accidentally using a banned substance. Pharmacists can also educate athletes and the public about the health consequences of using performance-enhancing substances. Pharmacists can play a variety of roles to assist with anti-doping. Such roles include educating, advising, dispensing and monitoring medications and supplements; and working with anti-doping agencies. There are few established educational opportunities for pharmacists and pharmacy students. Educational programs in sports pharmacy and doping control need to be developed for instruction in the classroom, for post-graduate training and for experiential programs. Classroom instruction should include information about performance-enhancing substances and general principles of doping control. Student activities for an established advanced pharmacy practice experience include education on performance-enhancing substances and assay technologies, preparing and providing presentations to athletes and others regarding these substances, performing literature research on drugs and dietary supplements used to improve athletic performance, writing a monograph on these substances, and participating in doping control programs.
The world constructed by self-organization of some amphiphils was discussed on the basis of micelle formation, vesicle formation, and oriented-nano-wire formation. First, the micelle formation of a both water- and oil- soluble surfactant, Aerosol OT, was discussed. Solution states of micelles and monomer were discussed on the basis of thermodinamic and NMR spectroscopic analyses of micelle formation. Next, micelle-vesicle transition was discussed. It was proposed that the phospholipid LUV formation by removing detergents and destruction by adding detergents occurred via 4 stages. The 4 stage model instead of the 3 stage model could not only elucidate the complicated phenomena observed during micelle-vesicle transition, but predicted the size and properties of the vesicles formed by detergent removal from mixed micelles. Next, the vesicle formation of a fatty acid with a single hydrophobic chain different from phospholipid, which has two hydorophobic chains, was discussed. The vesicle formation was strongly affected by the presence of preformed vesicles and the size was biased on the preformed vesicles. It was shown there exist two pass ways in the process of micelle-vesicle transition by pH jump. One is fission of the preformed vesicles after transfer of monomers from newly added oleate micelles and the other is transition from the mixed micelles after partial solubilization by the oreate micelles. Then, the vesicle formation of HCO-10, which has 3 hydrophobic chains, the mixed vesicle formation of phosphatidylethanolamine and lysophosphtidylcholine, which can not form vesicles, and the phospholipid vesicle formation and destruction by removing and adding PEG-lipid, were discussed. Lastly, oriented nano wire formation of mulamyldipeptid-conjugated lipids with ca 5 nm of diameter was discussed.
Aeromonas species are Gram-negative facultative anaerobic bacteria found ubiquitously in a variety of aquatic environments and most commonly implicated as causative agents of gastroenteritis. Sepsis is a fatal complication of Aeromonas infectious diseases, particularly in immune-compromised patients. Two species, Aeromonas hydrophila and Aeromonas sobria, are associated with human disease. Feasible virulence factors of Aeromonas include fimbrial and afimbrial adhesion molecules, hemolysins, enterotoxins, lipases and proteases. Recently, we purified a 65-kDa A. sobria serine protease (ASP) from the culture supernatant of A. sobria and found that the enzyme induces vascular leakage and reduces blood pressure through activation of the kallikrein/kinin system. This ASP activity potentially contributes to the onset of septic shock, suggesting ASP as an important virulence factor. In this review, I describe both the substrate specificity and the structural property of ASP. Furthermore, I also discuss the maturation mechanism of ASP. Further studies will facilitate development of novel drugs for bacterial infection that have inhibitory effects on various serine proteases.
Transcription is one of the most fundamental cellular functions and is an enzyme-complex mediated reaction that converts DNA sequences into mRNA. TATA-box is known to be an important motif for transcription. However, there are majority of promoters that have no TATA-box. They are called as TATA-less promoters and possess other elements that determine the transcription start site (TSS) of the genes. Multiple protein factors including ETS family proteins are known to recognize and bind to the GGAA containing sequences. In addition, it has been reported that the ETS binding motifs play important roles in regulation of various promoters. Here, we propose that the duplication and multiplication of the GGAA motifs are responsible for the initiation of transcription from TATA-less promoters.
To compare the difference of α-glucosidase inhibitory effect of total iridoid glycosides from crude products and processed products (Paozhi in Chinese) of Cornus officinalis Sieb. et Zucc. by α-glucosidase and glucose tolerance test in mice. Alcohol 40% was employed to extract the total iridoid glycosides from the crude products and the processed products then the total iridoid glycosides were enriched through macroporous resins to obtain SZY-1 and SZY-2 from the crude products and the processed products, respectively. The inhibitiory activity of these extracts was investigated on α-glucosidase inhibition in vitro and glucose tolerance test in vivo. The results demonstrated that both SZY-1 and SZY-2 were successfully enriched by macroporous resins. The contents of the total iridoid glycoside in SZY-1 and SZY-2 were 75.3% and 42.8%, respectively, and exerted inhibitory effects at various concentrations on α-glucosidase. The maximal inhibition ratio (86%) of SZY-1 was markedly higher than that of SZY-2. Meanwhile, SZY-1 was more potent than SZY-2 in decreasing blood sugar in diabetic mice induced by streptozotocin. Their effects were dependent on the dosage. In conclusion, the iridoid total glycoside from crude products and processed products of C. officinalis had α-glucosidase inhibitory effects in vitro and in vivo, and the effects of the crude products ware superior to those of processed products. It was also suggested that the hypoglycemic effects of C. officinalis may be attributable to its total iridoid glycoside.
Daidzein-loaded chitosan microspheres were prepared by emulsification/chemical-crosslinking technique. The dialysis bag method determined the release of daidzein from the microspheres. It demonstrated that the accumulative release curve in vitro was fit for the zero-order release equation and had good correlation with the absorptive fraction in vivo, suggesting the dialysis bag method evaluated the release of the microspheres well. The release of chitosan determined by the ninhydrin assay in vitro was very slow, less than 3 percent at 35 day. The pathological section by hematoxylin-eosin staining found the good biocompatibility of the prepared microspheres in the injective site. Combining the degradation photos by scanning electron microscopy with the plasma concentration-time data, it was speculated that the drug on the surface of the microspheres firstly released, then the major of drug near the surface and the inner of the microspheres released by diffusion through the shallow cavities and crack, lastly the drug released rapidly and completely being companied with the beginning of polymer degradation.
The local delivery of antibiotics in the treatment of infectious respiratory diseases is an attractive alternative to deliver high concentration of antimicrobials directly to the lungs and minimize systemic side effects. In this study, inhalable microparticles containing doxycycline hyclate, sodium carboxymethylcellulose, leucine and lactose were prepared by spray drying of aqueous ethanol formulations. Box-Behnken design was used to study the influence of various independent variables such as polymer concentration, leucine concentration, ethanol concentration and inlet temperature of the spray dryer on microparticle characteristics. The microparticles were characterized in terms of particle morphology, drug excipient interaction, yield, entrapment efficiency, Carr's index, moisture content, thermal properties, X-ray powder diffraction, aerosolization performance and in vitro drug release. The effect of independent variables on spray dryer outlet temperature was also studied. The overall shape of the particles was found to be spherical like doughnuts in the size range of 1.16-5.2 μm. The optimized formulation (sodium carboxymethylcellulose concentration 14% w/v, leucine concentration 33% w/v, ethanol concentration 36% v/v, inlet temperature of 140°C) exhibited the following properties: yield 56.69%, moisture content 3.86%, encapsulation efficiency 61.74%, theoretical aerodynamic diameter 3.11 μm and Carr's index 23.5% at an outlet temperature 77°C. The powders generated were of a suitable mass median aerodynamic diameter (4.89 μm) with 49.3% fine particle fraction and exhibited a sustained drug release profile in vitro.
This study examined the effects of nutrition support team (NST) services on nutrition supply type and patient outcomes in the intensive care unit (ICU) of a general hospital in South Korea. We retrospectively analyzed hospital records of patients who received either parenteral or enteral nutrition support during their ICU stays in the second half of NST (Nutrition Support Team) year 1 (2008) and NST year 2 (2009). Several measures of year 1 were compared with those of year 2, when more NST interventions were conducted. Number of nutrition prescriptions delivered to the ICU was 629 in year 1 and 677 in year 2. The increase in NST interventions led to a reduction in costly parenteral nutrition use, especially in surgical units. Number of patients selected for outcome measures was 40 in year 1 and 45 in year 2. There was an average 3.7 days reduction in the length of hospital stays between the two study terms. The average days of fasting were 3.3 days in year 1 and 1.3 days in year 2, which is statistically a significant decrease (p=0.02). The duration of parenteral nutrition decreased from 5.6 to 5.0 days as recommended. Compared with the amount of calories required, an average of 89.4% calories was delivered in year 1, and an average of 99.8% calories was delivered in year 2. Providing NST services in the ICU enhanced adequate nutrition support, cost savings, and better outcomes of the patients.
Kikisui® is a herbal lotion containing Kochia scoparia Fruit and Cnidium monnieri Fruit that is clinically used as an antipruritic for itchy dry skin. However, this formulation is unsuitable for inducing a prolonged effect. Here, we attempted to change the formulation from a lotion to a cream. The cream we chose was a water-in-oil (W/O) type emulsion for enhancing skin compatibility. In addition, the high water content imparts a sensation of coolness. However, it is difficult to prepare a stable W/O type cream with high water content using a mechanical mixing method. Instead, we prepared the W/O type emulsion using liquid crystals. Water containing cocamidopropyl betaine was added to a dispersed phase comprising an oil phase of oleic acid and liquid paraffin that was constantly stirred. Addition of an aqueous solution containing Kochia scoparia Fruit and Cnidium monnieri Fruit decreased the stability of the cream. However, addition of glycerin as a humectant, and ethyl p-hydroxybenzoate/n-butyl p-hydroxybenzoate as preservatives enhanced the stability of the cream. The stability of the emulsion was correlated with the apparent viscosity of the cream. The final W/O type cream had a water content of 83% and was stable for more than 6 months at 4°C. Furthermore, ostol, which is one of the main biologically active herbal compounds, was also stable for more than 6 months.
5-Lipoxygenase (5-LOX), which is believed to be a major source of oxidative stress, participates in somatostatin-receptor transmembrane signaling in the central nervous system. We used the Tet-On inducible expression system in PC12 cells to obtain cell lines with reproducible, stable 5-LOX expression levels to study its function. Cell apoptosis rates induced by Aβ42 were determined using an apo-BrdDU kit. Lipid peroxide, antioxidant enzyme, and caspase-3 activities were evaluated with respective commercial kits. The expression of 5-LOX, bcl-2, and bax were detected by immunoblotting. A subclone of PC18 with Tet-On inducible expression of 5-LOX was selected from PC12 transfectants. Expression of 5-LOX had no significant inhibitory effect on the cell viability of the PC18 clone. In contrast, compared with the control group, the cell viability of clone PC18 was significantly reduced after the induction of 5-LOX during Aβ exposure. The differences in cell viability before and after the induction of 5-LOX during Aβ insult were significantly offset by AA861. Overexpression of 5-LOX only slightly improved the activities of superoxide dismutase (SOD). The levels of intracellular peroxides, SOD and caspase-3 activity, and Bax expression were significantly upregulated, and the levels of glutathione peroxidase and catalase were downregulated correspondingly in clone PC18 during Aβ exposure. These results indicate that constitutive expression of 5-LOX is not detrimental per se, but overexpression of 5-LOX may become problematic during Aβ exposure.