Motofumi Hiyoshi

DNA extraction from human urinary sediment

DNA was extracted from urinary sediments and was sufficient for polymerase chain reaction (PCR) and enzymatic analysis, even if DNA from microorganisms coexisted. From urine samples, the yield of DNA ranged from trace levels to 20 μg per 10 mL urine. When urinary sediment was stored in ethanol, DNA remained stable for 2 weeks or more. Individual identification and sex determination could easily be performed using either fresh or ethanol‐fixed urine. In conclusion, urine can be used as a source for PCR‐based investigations and genetic studies. J. Clin. Lab. Anal. 12:88–91, 1998.

A Pedigree with Pendred Syndrome: Case Report and Discussion on Hereditary Hearing Loss

After presenting the case of a Japanese pedigree with Pendred syndrome we discuss whether congenital hearing loss such as Pendred syndrome can be avoided.

どんな基礎疾患がサイトメガロウイルス感染を引き起こしやすいか? 初めての標準化された客観的サイトメガロウイルス用PCRキットを使用した, 良性疾患, 血液悪性腫瘍, 骨髄悪性腫瘍, 骨髄移植後, 腎移植後における比較

All immunocompromised hosts, such as infants, the elderly, patients with advanced cancer, and patients being treated with immunosuppressants, etc., are said to be more susceptible to cytomegalovirus (CMV) infection or CMV disease. However, we questioned the validity of this conclusion and attempted to detect CMV viremia in the plasma of subjects by using the AMPLICOR CMV test (Roche Diagnostics Systems, Branchburg, NJ), the first standardized PCR kit for CMV infection. One hundred healthy volunteers whose CMV IgG titer was < 4 and 100 healthy volunteers whose IgG titer for CMV was > or = 4 were studied. None of the subjects in either healthy group was positive for CMV viremia. Patients who were suspected of CMV infection were divided into four groups and tested: [1] 104 patients with benign disease, only one of whom was positive for CMV [2] 99 patients with hematopoietic malignancy who had not undergone bone marrow transplantation and all of whom were negative for CMV infection [3] 120 post-bone-marrow transplantation, 28 of whom were CMV positive, [4] 37 post-renal transplantation patients, 19 of whom were CMV positive. A statistically significant difference in CMV positivity was found by the non-parametric Kruskal-Wallis test (p < 0.0001) among the four disease group. CMV infection has been said to occur in all types of immunocompromised patients, however, based on our findings, we conclude that CMV infection tends to occur in post-transplantation status and does not tend to occur in patients with hematopoietic malignancy if they have not undergone transplantation.

Interleukin-2 induces motility of human lymphocytes, but not their mobility

Interleukin-2 induces cytotoxic and antitumor activities of human lymphocytes. For the expression of these activities, the cytoskeletal system is probably activated. This study was do;ne to find if interleukin-2 causes cell movement. Lymphocytes were incubated with interleukin-2, and their morphology and motile activities were studied. After 72 hr of incubation, some 29% of lymphocytes were larger than before; nucleoli had formed and the microvilli were well-developed. The membrane potential of the lymphocytes increased during incubation. Motility under agar after 3 days of incubation with interleukin-2 was examined. Cells aggregated in clumps in the incubation well, and migration was not observed. When mobility was examined with Boydens method, fewer cells incubated with interleukin-2 migrated than in control preparations. Cells incubated with interleukin-2 were extracted with Triton X-100. The extract obtained had three more bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis than the controls. The band were at the positions for the molecular weights of 270,000-300,000. We concluded that interleukin-2 activated the motility of lymphocytes, but not their mobility.