著者

菊池 真 舘 延忠 小塚 直樹 二宮 孝文 小林 正裕 堀本 佳誉 内田 英二 佐々木 公男 辰巳 治之

出版者

札幌医科大学医学部

雑誌

札幌医学雑誌 = The Sapporo medical journal = The Sapporo medical journal (ISSN:0036472X)

巻号頁・発行日

vol.74, no.3, pp.39-44, 2005-08-01

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Formalin is a main fixative in the field of pathology. Molecular biological analysis of formalin-fixed samples was difficult because formalin fixation decreased the quality of isolated DNA. Therefore, we compared the quality of DNA obtained by using DNA extraction kit (Sepa GeneR) to that using proteinase K. Using proteinase K, it was possible to extract high quality DNA, and obtain DNA from samples of 3 months fixative. Moreover, by proteinase K method, it was also possible to analyze aprataxin gene exon 5 in DNA extraction from formalin-fixed human brain tissues from a suspected case of early-onset ataxia with ocular motor apraxia and hypoalbuminemia (EAOH). The aprataxin gene exon 5 DNA sequences were obtained following in vitro gene amplification using nested-PCR. Mutation on aprataxin gene exon5 was not observed in the suspected case of EAOH; however, it was possible to perform sequence analysis of aprataxin gene exon5. This method was more useful for DNA extraction and direct sequencing of formalin-fixation samples than the kit method.

著者

績 晶子

出版者

札幌医科大学医学部

雑誌

札幌医学雑誌 = The Sapporo medical journal = The Sapporo medical journal (ISSN:0036472X)

巻号頁・発行日

vol.55, no.6, pp.545-564, 1986-12-01

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The combination of high doses of hepatitis B immunoglobulin (HBIG) and hepatitis B vaccine (HB vaccine) were injected for the prevention of maternal infant transmission of hepatitis in 185 infants whose mothers were hepatitis B carriers. Except for 9 patients who had intrauterine infection before birth, 176 infants recieved the first injection of HBIG within 48 hours after birth. These infants were protected from any perinatal transmission, and later underwent HB vaccination. As a results, the 176 cases were classified into a group of good responders, poor responders and non responders. The good responders were the infants who acquired the anti-HBs antibody easily by vaccination, and the poor or non responders were the infants who had difficulty in producting the anti-HBs antibody inspite of frequent vaccinations. This study was to compare the group of poor or non responders with the group of good responders in terms of some of the cellular immunity. Results: 1) Poor or non responders could be infected with HBV due to the lack of active immunization. In order to prevent the HBV infection in poor or non responders, continuation of HBIG injection was required every two to three months. 2) In vitro lymphoproliferative responses to PHA-P, Con-A, PWM, PPD or Cytomegalovirus antigen in peripheral lymphocytes were normal in all three groups. The response to HBsAg was significantly decreased in the poor or non responder groups. 3) The effector cells for HBsAg consisted mainly of T lymphocytes. These were classified as the T4 subset (helper/inducer T) which expressed TAC antigen (IL-2 receptor) by the stimulation with HBsAg. 4) From these results, the deficient production of the anti-HBs antibody and the low response to in vitro lymphoproliferative activity to HBsAg in poor or non responders were specific immunological dysfunctions for HBsAg.

著者

山内 一功 太田 勲

出版者

札幌医科大学医学部

雑誌

札幌医学雑誌 = The Sapporo medical journal = The Sapporo medical journal (ISSN:0036472X)

巻号頁・発行日

vol.47, no.6, pp.582-591, 1978-12-01

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Changes of mechanical and electrical responses in frog whole sartorius muscle surviving at a constant temperature (20±1℃) in normal Ringer solution, Ringer solution which was supplemented with antibiotics or modified Medium 199 were examined and the following results were obtained. 1. The peak tensions of both twitch and potassium contracture (K-contracture) decreased gradually with the lapse of time after immersion of whole sartorius muscles in normal Ringer solution. Twelve hours after the immersion, the peak tensions of twitch and K-contracture decreased to about 55% and 67% of the control, respectively. Sixteen hours after the immersion, the respective peak tensions decreased to about 10% of the control. However, the peak tension of caffeine contracture did not change up to 12 hours after immersion, whereas it decreased to about 20% of the control 16 hours after immersion. Whole sartorius muscles surviving more than 12 hours in normal Ringer solution assumed a milky color and contained fibers which had a granular appearence. Proliferation of bacteria was observed in normal Ringer solution in which muscle preparations survived more than 12 hours. 2. In whole muscle surviving in Ringer solution with antibiotics (penicillin 50U/ml plus streptomycin 10 μg/ml), the twitch tension decreased gradually with the lapse of time after the immersion and it decreased to about 10% at 24 hours after immersion. On the other hand, the time course of the inhibition of the magnitude of K-contracture tension showed two distinct phases ; the first phase in which the tension was inhibited very slowly and slightly (up to 36 hours after immersion), and the second phase in which tension was inhibited rapidly and completely. In addition, peak tension of caffeine contracture showed no changes up to 24 hours after the immersion whereas it decreased to about 23% of the control 48 hours after the immersion. 3. The magnitude of resting potential and the amplitude of action potential of fibers immersed in Ringer solution with antibiotics for 24 hours were quite similar to those of the control. Fourty-eight hours after the immersion, overshoot of action potential was not observed, although the magnitude of resting potential showed no changes. 4. Twitch was abolished 6 days after immersion of whole muscle in modified Medium 199, whereas K-contracture and caffeine contracture were observed even after 18 days of the immersion. On the basis of these results, the mechanism of inhibition of mechanical responses in surviving whole muscle was discussed.