In treating diabetes over the long term, controlling the daily life is very important and cannot be neglected. In particular, the patient’s awareness and practice of their treatment had a great influence on their treatment. However, there has been very little scientific research done with regard to how much the patient understands the instructions about taking their medicine and whether they actually follow those instructions. Due to these facts, in this research we studied what guidance should be given when taking medicines made from insulin after analyzing the results of an investigation taken from the patients’ point of view. In carrying out a study on the education and the degree of understanding of insulin selfinjections, it became clear that older patients’ ability to understand diminished, and the patients forgot to shake the NPH insulin immediately before injection. And, in more than a few cases, even when the NPH insulin was shaken, injections took place after some time had passed. After studying the amount of time that elapses once sedimentation begins, it was found that the insulin is effective if it is injected within 2.5 minutes of being shaken. Next, a study was carried out on the problems of maintaining the quality of life (QOL) of patient while continuing the treatment for diabetes. As a result, it became clear that the patient’s lifestyle, including psychological factors, exerts a large influence on continuing the correct treatment. In particular, in cases of insulin injections when the patient eats out, younger patients tend to be more susceptible to psychological influence. This suggests that there is a need to work on guidance for the patient in taking their medicine. And, such guidance should strictly adhere to information regarding the patient’s lifestyle, which comes through good communication with the patient. On the other side, the coring of the insulin vial of insulin was examined in terms of medicine production technology. As a result, the occurrence of coring is seen irrespective of the type of needle and the temperature of the rubber vial stopper, and the fact that rubber fragments were found in the injection solution, suggesting the possibility of subcutaneous rubber fragments through injection. Further examination of the problems of continued long-term treatment of injected medicines may be necessary in the future. Due to the necessity of self-control of patients and the long time span of treatment that is involved, maintaining the QOL of the patients is important in diabetes treatment. We feel that it is important to give guidance about treatment, which fits the patients’ lifestyles.
Astrocytes, the most abundant glial cell type in the brain, are considered to have physiological and pathological roles in neuronal activities. We found that reperfusion of cultured astrocytes after Ca2+ depletion causes Ca2+ overload followed by delayed cell death and the Na+-Ca2+ exchanger in the reverse mode is responsible for this Ca2+-mediated cell injury (Ca2+ paradox injury). The Ca2+ paradox injury of cultured astrocytes is considered to be an in vitro model of ischemia/reperfusion injury, since a similar paradoxical change in extracellular Ca2+ concentration is reported in ischemic brain tissue. This review summarizes the mechanisms underlying the Ca2+-mediated injury of astrocytes and the protective effects of drugs against Ca2+ reperfusion injury. This study shows that Ca2+ reperfusion injury of astrocytes is accompanied by apoptosis as evidenced by DNA fragmentation and nuclear condensation. Calpain, reactive oxygen species, calcineurin, caspase-3, and NF-κB are involved in Ca2+ reperfusion-induced delayed apoptosis of astrocytes. Several drugs including CV-2619, T-588 and ibudilast protect astrocytes against the delayed apoptosis. CV-2619 prevents astrocytes from the delayed apoptosis by production of nerve growth factor, resulting in an activation of mitogen-activated protein (MAP)/extracellular signal-regulated kinase (ERK) and phosphatidylinositol-3 (PI3) kinase signal pathways. The protective effect of T-588 is mainly mediated by an activation of MAP/ERK signal cascade. Moreover, ibudilast prevents the Ca2+ reperfusion-induced delayed apoptosis of astrocytes via cyclic GMP signaling pathway. Further studies in this system will contribute to the development of new drugs that attenuate ischemia/reperfusion injury via modulation of astrocytes.
A body of evidence supports that excitatory amino acid systems, particularly glutamatergic one, participate in morphine dependence and naloxone-precipitated withdrawal. In this study, we examined the involvement of glial glutamate transporters, GLT-1 and GLAST, in them. Rats were rendered morphine-depndent by subcutaneous implantation of two 75 mg morphine pellets for 5 days. Intracerebroventricular administration of DL-threo-β-benzyloxyaspartate, a glutamate transporter inhibitor significantly facilitated various naloxone-precipitated withdrawal signs. By northern blot analysis, the expression of GLT-1 mRNA was found to decrease significantly in the striatum and thalamus of morphine-dependent rats, and to increase significantly in the striatum 2 hr after the naloxone-precipitated withdrawal. On the other hand, there were no significant changes in GLAST mRNA levels in any brain regions. In vivo microdialysis experiments revealed that the extracellular glutamate levels was elevated in the striatum and nucleus accumbens, in which the changes of GLT-1 mRNA level were observed, during naloxone-precipitated morphine withdrawal. In cultured astrocytes, the expression of GLT-1 mRNA was regulated by agents activating the cAMP pathway, as well as β-adrenergic agonist and dopamine, but not morphine. These results suggest that the changes of GLT-1 expresson, which alter the glutamate uptake and affect the glutamatergic transmission efficiency, play a role in the development of morphine dependence and the expression of morphine withdrawal.
The new way with nevirapine was reported to prevent mother-to-child transmission of the AIDS virus (HIV) that also is less costly and markedly more effective than the standard therapy with AZT in the third world. The more practical therapy comes from substituting one marketed drug, nevirapine, for the standard drug, AZT. It was proposed that widescale use of nevirapine in developing countries could potentially prevent 300000 to 400000 newborns each year from beginning life infected with HIV. But in September, 2000, the Center for Disease Control and Prevention (CDC) received reports of life-threatening hepatotoxicity (liver damage) among health care workers taking nevirapine for post-exposure prophylaxis (PEP) after occupational exposure to HIV. Furthermore, persons taking nevirapine regimens for PEP after HIV exposure also are at risk for serious and adverse events. The federal Food and Drug Administration (FDA) had identified 22 cases of severe liver, skin and muscle damage related to nevirapine taken after possible exposure to HIV from March 1997 through September 2000. Nevirapin has not been recommended for PEP use, and has previously been associated with instances of serious skin or muscle conditions, liver damage, and death when used for treating HIV-infected individuals. In most circumstances, the risks associated with nevirapine as part of an occupational PEP regimen might outweigh the anticipated benefits.
A prior review system for clinical trials was introduced into The University of Tokyo Hospital in March 1999, for the purpose of supporting the Institutional Review Board (IRB). In this study, we investigated the effect of this system on IRB, and evaluated roles of pharmacists involved in utilizing this prior review system. An average period from the acceptance of regular application of each protocol to the approval of IRB was 1.7 months, and the period was significantly shorter than that before the introduction of this system (2.3 months). On the other hand, the number of instructions given to applicants increased to be 2.8 times before the introduction of this system, suggesting the improvement of the quality of this prior review system. In the prior review, the number of instructions pointed out from pharmacists concerning pharmacokinetics, pharmacodynamics, drug-interactions, adverse reactions, and the other matters related to investigational drugs was greater than that from doctors or nurses. According to the results of the surveys for doctors, nurses, and other clerical employees, pharmacists were indicated to review from a pharmaceutical point of view and their review was found to be very important. In conclusion, the prior review system was considered to be useful for the rational practice of clinical trials and pharmacists were recognized to be indispensable as reviewers on the prior review.
The in vitro effect of various glycosaminoglycans (GAGs) on the clonal growth of CD34+ megakaryocytic progenitor cells (CFU-Megs) isolated from human placental/umbilical cord blood (CB) was evaluated in human plasma containing semisolid culture stimulated by recombinant human thrombopoietin (TPO). The GAGs,including hyaluronic acid from human umbilical cords (HA-h), pig skins (HA-p) and rooster combs (HA-r), or keratan sulfate (KS), various chondroitin sulfates (CS-A, B, C, D, E), and heparan sulfate (HS), were tested. Each GAG alone did not affect the clonal growth of CFU-Meg. In the presence of TPO, adding of HA-p or HS (100 μg/ml) resulted in an approximately 1.3-fold increase, in the total number of colonies, due to an increase in large megakaryocyte colonies. In contrast, CS-E led to a marked decrease in CFU-Meg growth. At the end of the culture, the total number of cells increased 3.0-fold of the initial value of the control, but adding HA-p or HS showed an approximately 9.1-fold or 18.3-fold increase. Similarly, the total number of CFU-Meg detected in the harvested cells increased to 4.8-fold of the initial value, while, an approximately 18.3-fold or 38.8-fold increase was observed in the culture containing HA-p or HS, respectively. Flow cytometric analysis of the harvested cells showed no significant difference in the expression of surface antigens and DNA ploidy distribution of megakaryocytes between the control and GAG treatments. These results suggest that HA-p and HS promote the proliferation of immature CB CD34+ CFU-Meg in the presence of TPO.
Direct sequencing using λphage DNA and E.coli colonies with plasmid DNA is a very powerful technique. Almost all of the reported direct sequencing methods involve either radioactive sequencing or fluorescent dye-primer sequencing. We present a direct colony sequencing strategy that uses a dye terminator (BigDye terminator kit) together with dyeprimer sequencing. We found that single-colony sequencing with the terminatory yielded about 500 base pairs of sequence information. Signal strength was not improved when the number of cycles increased to 40. The colony used for the sequencing was estimated to contain about 5.6×107 cells. In addition, although a single plaque consisted of 2×106 cells, the pfu was not high enough to read with single-cycle sequencing, and only about 300 base pairs of sequence information were obtained from a single plaque using two cycle-sequencing reactions (re-cycle sequencing). The optimal amounts of the template were 500 ng of purified λDNA and 1×107 pfu of the λphage suspension, but with BigDye™ terminator it was possible to detect as little as 50 ng of purified λDNA and 2×106 pfu for λphage suspensions. Thus, colony direct sequencing and plaque direct sequencing are estimated to be very useful for rapid and high-throughput screening of genomic and cDNA libraries.