Muscle plasticity related to changes in tubulin and αB-crystallin levels induced by eccentric contraction in rat skeletal muscles.

We used the model of eccentric contraction of the hindlimb muscle by Ochi et al. to examine the role of eccentric contraction in muscle plasticity. This model aims to focus on stimulated skeletal muscle responses by measuring tissue weights and tracing the quantities of αB-crystallin and tubulin. The medial gastrocnemius muscle (GCM) responded to electrically induced eccentric contraction (EIEC) with significant increases in tissue weight (p < 0.01) and the ratio of tissue weight to body weight (p < 0.05); however, there was a decrease in soleus muscle weight after EIEC. EIEC in the GCM caused contractile-induced sustenance of the traced proteins, but the soleus muscle exhibited a remarkable decrease in α-tubulin and a 19% decrease in αB-crystallin. EIEC caused fast-to-slow myosin heavy chain (MHC) isoform type-oriented shift within both the GCM and soleus muscle. These results have shown that different MHC isoform type-expressing slow and fast muscles commonly undergo fast-to-slow type MHC isoform transformation. This suggests that different levels of EIEC affected each of the slow and fast muscles to induce different quantitative changes in the expression of αB-crystallin and α-tubulin.

Improvement of foaming property of egg white protein by phosphorylation through dry-heating in the presence of pyrophosphate.

Egg white protein (EWP) was phosphorylated by dry-heating in the presence of pyrophosphate at pH 4 and 85 degrees C for 1 d, and the foaming properties of phosphorylated EWP (PP-EWP) were investigated. The phosphorus content of EWP increased to 0.71% as a result of phosphorylation. To estimate the foaming properties of EWP, the foams were prepared by 2 methods: bubbling of the 0.1% (w/v) protein solution and whipping of the 10% (w/w) protein solution with an electric mixer. The foaming power, which was defined as an initial conductivity of foam from 0.1% (w/v) protein solution, was a little higher in PP-EWP than in native EWP (N-EWP), and the foaming stability of PP-EWP was much higher than that of dry-heated EWP (DH-EWP) and N-EWP. The microscopic observation of foams from the 10% (w/w) solution showed that the foams of PP-EWP were finer and more uniform than those of N- and DH-EWP. Although there were no significant differences in the specific gravity and overrun of the foams between PP- and DH-EWP (P < 0.05), the specific gravity and overrun of the foams from PP-EWP were smaller and higher, respectively, than that of the foams from N-EWP. The drainage volume was smaller in the foams from PP-EWP than in those from N- and DH-EWP. These results demonstrated that phosphorylation of EWP by dry-heating in the presence of pyrophosphate improved the foaming properties, and that it was more effective for the foam stability than for the foam formation.

Relaxation and immunity enhancement effects of gamma-aminobutyric acid (GABA) administration in humans.

The effect of orally administrated gamma-aminobutyric acid (GABA) on relaxation and immunity during stress has been investigated in humans. Two studies were conducted. The first evaluated the effect of GABA intake by 13 subjects on their brain waves. Electroencephalograms (EEG) were obtained after 3 tests on each volunteer as follows: intake only water, GABA, or L-theanine. After 60 minutes of administration, GABA significantly increases alpha waves and decreases beta waves compared to water or L-theanine. These findings denote that GABA not only induces relaxation but also reduces anxiety. The second study was conducted to see the role of relaxant and anxiolytic effects of GABA intake on immunity in stressed volunteers. Eight acrophobic subjects were divided into 2 groups (placebo and GABA). All subjects were crossing a suspended bridge as a stressful stimulus. Immunoglobulin A (IgA) levels in their saliva were monitored during bridge crossing. Placebo group showed marked decrease of their IgA levels, while GABA group showed significantly higher levels. In conclusion, GABA could work effectively as a natural relaxant and its effects could be seen within 1 hour of its administration to induce relaxation and diminish anxiety. Moreover, GABA administration could enhance immunity under stress conditions.

Suppressive effect of functional drinking yogurt containing specific egg yolk immunoglobulin on Helicobacter pylori in humans.

Helicobacter pylori is a human pathogen that infects over 50% of the population worldwide. It is the most important etiologic agent of gastroduodenal ulcers and malignancies. Helicobacter pylori urease enzyme is considered the main factor for the organism's colonization in the gastroduodenal mucosa. Hens immunized with the purified urease produce a highly specific anti-H. pylori urease immunoglobulin (IgY-urease) in their egg yolks. Immunoglobulin Y-urease was stable at 60 to 65 degrees C for 30 min and at pH 4.0 for 7 h. Its activity was lost at 80 degrees C for 20 min and at pH 2 for 4 h. Specially designed functional drinking yogurt containing Lactobacillus acidophilus and Bifidobacterium spp. with 1% egg yolk IgY-urease was produced commercially. Immunoglobulin Y-urease activity showed stability in the product up to 7 d, and then decreased to 85% after 3 wk of storage. A clinical study was conducted to determine the effectiveness of IgY-urease yogurt to suppress infection in humans. Forty-two volunteers who tested positive for H. pylori using a 13C-urea breath test were recruited. A total of 450 mL of IgY-urease (test group) or IgY-urease-free yogurt (control group) was consumed in 150-mL portions 3 times daily for 4 wk. Volunteers were tested after 2 and 4 wk; urea breath test values significantly decreased in the test group compared with the control group. The results indicate that suppression of H. pylori infection in humans could be achieved by consumption of drinking yogurt fortified with IgY-urease.

Comparison of 5-fluorouracil and 5-fluoro-2'-deoxyuridine as an effector in radiation-activated prodrugs.

The purpose of this study was to clarify whether 5-fluoro-2'-deoxyuridine (FdUrd) is superior to 5-fluorouracil (5-FU) as an effector in the radiation-activated prodrugs which we have been developing. The in vitro cytotoxicity of 5-FU and FdUrd was compared in two murine tumor and four human pancreatic cancer cell lines using a colony assay and in vivo efficacy was compared with SCCVII tumor using a growth delay time assay. FdUrd was slightly more hydrophilic than 5-FU. In vitro, FdUrd was more efficient than 5-FU in two lines, whereas 5-FU was more efficient in two lines and the two drugs were almost equal in efficacy in the remaining two. The concentration to reduce tumor cell survival to 50% after 24-h drug exposure was 5-32 microM for both 5-FU and FdUrd in murine lines, while it was 30-210 microM in human pancreatic cancer cell lines. The difference in relative efficacy of the two drugs among these cell lines could not be attributed to the rate of intracellular uptake of the compounds. FdUrd was less toxic than 5-FU in C3H/He mice, and FdUrd was less efficient than 5-FU in SCCVII tumors in vivo. These results suggest that FdUrd is not necessarily more potent than 5-FU, and development of the FdUrd prodrugs may not necessarily turn out to be fruitful.

Thermostabilization of ovalbumin in a developing egg by an alkalinity-regulated, two-step process.

Native ovalbumin has been known to convert into a heat-stable form, S-ovalbumin, either in an avian shell egg or in an isolated ovalbumin solution. Recently, similar conversion of ovalbumin in fertile eggs was also reported. We found that the conversion into S-ovalbumin was slower in fertile eggs than in unfertile eggs under the same incubation conditions on the basis of calorimetric analyses for the samples isolated from those eggs. During the incubation, there were differential pH changes of white in the fertile and unfertile eggs. When the pH of purified ovalbumin was manually adjusted so as to simulate the pH changes of egg white during the incubation, the course of the conversion into S-ovalbumin was very similar to that either in fertile or unfertile eggs. Therefore, we conclude that thermostabilization of ovalbumin in fertile eggs proceeds by a certain mechanism which depends on the alkalinity of egg white.

Tissue-specific and isoform-specific changes in MCT1 and MCT4 in heart and soleus muscle during a 1-yr period.

We examined the postnatal changes (days 10, 36, 84, 160, 365) of monocarboxylate transporters (MCT)1 and MCT4 in rat heart and soleus muscle. In the heart, MCT1 protein and mRNA remained unaltered from day 10 until 1 yr of age. Both MCT4 protein and mRNA in the heart were detected at 10 days of age, but the MCT4 protein and transcript were not detected thereafter. In the soleus muscle, MCT1 protein (+38%) and mRNA (+136%) increased during the first 84 days and remained stable until 1 yr of age. In contrast, soleus MCT4 protein decreased by 90% over the course of 1 yr, with the most rapid decrease (-60%) occurring by day 84 (P < 0.05). At the same time, MCT4 mRNA was increased by 74% from days 10 to 84 (P < 0.05), remaining stable thereafter. In conclusion, developmental changes in MCT transport proteins are tissue specific and isoform specific. Furthermore, it appears that MCT1 expression in the heart and MCT1 and MCT4 expression in the soleus are regulated by pretranslational processes, whereas posttranscriptional processes regulate MCT4 expression in the soleus muscle.

Decreased monocarboxylate transporter 1 in rat soleus and EDL muscles exposed to clenbuterol.

We hypothesized that a shift in muscle fiber type induced by clenbuterol would change monocarboxylate transporter 1 (MCT1) content and activity of lactate dehydrogenase (LDH) and isoform pattern and shift myosin heavy chain (MHC) pattern in soleus (Sol) and extensor digitorum longus (EDL) of male rats. In the clenbuterol-administered rats (2.0 mg x kg(-1) x day(-1) subcutaneously for 4 wk), the ratio of muscle weight to body weight increased in the Sol (P < 0.05) and the EDL (P < 0.01). Clenbuterol induced the appearance of fast MHC(2D) and decreased slow MHC(1) in Sol (13%) but had no effect on EDL. The MHC pattern of Sol changed from slow to fast type. Clenbuterol increased LDH-specific activity (P < 0.01) and the ratio of the muscle-type isozyme of LDH to the heart type (P < 0.05) in Sol. The LDH total activity of the EDL muscle was also increased (P < 0.05). Furthermore, MCT1 content significantly (P < 0.05) decreased in both Sol and EDL (27 and 52%, respectively). This study suggests that clenbuterol might mediate the shift of MHC from slow to fast type and the changes in the regulation of lactate metabolism. Novel to this study is the observation that clenbuterol decreases MCT1 content in the hindlimb muscles and that the decrease in MCT1 is not muscle-type specific. It may suggest that the genetic expressions of individual factors involving slow-type MHC, heart-type isozyme of LDH, and MCT1 are associated with one another but are regulated independently.

N(1)-C(5')-linked dimer hydrates of 5-substituted uracils produced by anodic oxidation in aqueous solution.

Electrochemical dimerization reactivity has been studied for 5-substituted uracils (5XU) including thymine (1a: X = Me) and 5-halouracil derivatives (1b: X = F; 1c: X = Cl; 1d: X = Br; 1e: X = I). Upon galvanostatic electrolysis of Ar-saturated aqueous solution 1a underwent anodic oxidation to produce N(1)-C(5')- and N(1)-C(6')-linked dimer hydrates, 1-(6'-hydroxy-5',6'-dihydrothymin-5'-yl)thymine (5a) and 1-(5'-hydroxy-5',6'-dihydrothymin-6'-yl)thymine (6a), as the major products. These N-C-linked dimerizations were accompanied by the formation of novel stereoisomeric C(5)-C(5')-linked dimers (meso isomer: 13a[meso]; racemic isomer: 13a[rac]) with a condensed tetrahydrofuran ring skeleton. Similar electrolyses of 5-fluorouracil (1b) and 5-chlorouracil (1c) also afforded the corresponding N(1)-C(5')-linked dimer hydrates, 1-(5'-fluoro-6'-hydroxy-5',6'-dihydrouracil-5'-yl)-5-fluorouracil (5b) and 1-(5'-chloro-6'-hydroxy-5',6'-dihydrouracil-5'-yl)-5-chlorouracil (5c), respectively, while resulting in neither N(1)-C(6')-linked dimer analogues nor C(5)-C(5')-linked dimers, unlike the reactivity of 1a. In contrast to 1a-c, no dimeric products were obtained from 5-bromouracil (1d) and 5-iodouracil (1e). The present electrochemical method was applicable to the cross-dimerization into N(1)-C(5')-linked heterodimer hydrates composed of binary 5-substituted uracils that occurred in competition with the formation of homodimer hydrates. A mechanism of the N(1)-C(5')-linked dimerization of 1a-c has been proposed, by which allyl-type radical intermediates with limiting mesomeric forms of N(1)-centered and C(5)-centered pyrimidine radicals (2a-c [N(1)]/2a-c [C(5)]) are generated via anodic one-electron oxidation and subsequent deprotonation at N(1) and undergo a head-to-tail coupling.

In vivo evaluation of a novel antitumor prodrug, 1-(2'-oxopropyl)-5-fluorouracil (OFU001), which releases 5-fluorouracil upon hypoxic irradiation.

PURPOSE: We previously proposed that a prodrug of 5-fluorouracil (5-FU), OFU001, is activated through capturing of hydrated electrons produced by hypoxic irradiation. Because hydrated electrons are readily deactivated by oxygen, the 5-FU release occurs specifically upon hypoxic irradiation. In this study, we investigated the in vivo efficacy, pharmacokinetics, and toxicity of OFU001. METHODS AND MATERIALS: Female 10-week-old C3H/He mice bearing SCCVII tumors were used. To measure release of 5-FU from OFU001 in vivo, the mice were given 100 mg/kg of OFU001 intraperitoneally and irradiated. Thereafter, 5-FU levels in the tumor and serum were measured by high-performance liquid chromatography. To evaluate in vivo efficacy, OFU001 was administered 30 min before irradiation, and radiation-potentiating effects were investigated by means of a tumor growth delay assay and a 50% tumor control dose (TCD-50) assay. The lethal dose of OFU001 was evaluated in the same mice. RESULTS: Following administration of OFU001 and irradiation at 30 Gy, the average 5-FU levels in the tumor and serum were 179 ng/g and 83 ng/mL, respectively. Administration of OFU001 (100-200 mg/kg) to the tumor-bearing mice before a single dose of 15-Gy irradiation produced a mean tumor growth delay of 1-5 days as compared to radiation alone (although the delay was not significant). However, no additional growth delay was observed when OFU001 was combined with 5 radiation fractions of 4 Gy each. The enhancement ratio of OFU001 in the TCD-50 assay was 1.2. No mice died after administration of 0.6-1.2 g/kg of OFU001. CONCLUSIONS: OFU001 appears to work in vivo via the proposed mechanism of activation. Although the in vivo effect of this compound was not strong enough for clinical efficacy, these results should encourage further research on the development of prodrugs of more potent anticancer agents activated through the same mechanism.

Stereoelectronic effect on one-electron reductive release of 5-fluorouracil from 5-fluoro-1-(2-oxocycloalkyl)uracils as a new class of radiation-activated antitumor prodrugs.

A series of 5-fluoro-1-(2'-oxocycloalkyl)uracils (3-11) that are potentially novel radiation-activated prodrugs for the radiotherapy of hypoxic tumor cells have been synthesized to evaluate a relationship between the molecular structure and the reactivity of one-electron reductive release of antitumor 5-fluorouracil (1) in anoxic aqueous solution. All the compounds 3-11 bearing the 2'-oxo group were one-electron reduced by hydrated electrons (eaq-) and thereby underwent C(1')-N(1) bond dissociation to release 5-fluorouracil 1 in 47-96% yields upon radiolysis of anoxic aqueous solution, while control compounds (12, 13) without the 2'-oxo substituent had no reactivity toward such a reductive C(1')-N(1) bond dissociation. The decomposition of 2-oxo compounds in the radiolytic one-electron reduction was more enhanced, as the one-electron reduction potential measured by cyclic voltammetry in N,N-dimethylformamide became more positive. The efficiency of 5-fluorouracil release was strongly dependent on the structural flexibility of 2-oxo compounds. X-ray crystallographic studies of representative compounds revealed that the C(1')-N(1) bond possesses normal geometry and bond length in the ground state. MO calculations by the AM1 method demonstrated that the LUMO is primarily localized at the pi* orbital of C(5)-C(6) double bond of the 5-fluorouracil moiety, and that the LUMO + 1 is delocalized between the pi* orbital of 2'-oxo substituent and the sigma* orbital of adjacent C(1')-N(1) bond. The one-electron reductive release of 5-fluorouracil 1 in anoxic aqueous solution was presumed to occur from the LUMO + 1 of radical anion intermediates possessing a partial mixing of the antibonding C(2')=O pi* and C(1')-N(1) sigma* MO's, that may be facilitated by a dynamic conformational change to achieve higher degree of (pi* + sigma*) MO mixing.

Thymine carboxylation: nucleophilic addition of carbon dioxide radical anion.

PURPOSE: The nucleophilic addition properties of carbon dioxide radical anion (CO2*-) towards N1-substituted thymine derivatives in aqueous solution is studied for comparison with their one-electron reducing reactivity. MATERIAL AND METHODS: N2O-Saturated aqueous solutions of 1-methylthymine, 1,3-dimethylthymine, and thymidylyl(3'-->5')-thymidine containing excess formate ions were gamma-irradiated at 1.0Gy min(-1). Several carboxylated thymines were isolated by preparative HPLC and identified by GC-MS, NMR and X-ray crystallography. RESULTS: Along with one-electron reduction yielding N-substituted 5,6-dihydrothymines and C5--C5'-linked dihydrothymine dimers, the addition of CO2 radical anion(s) to the C5--C6 double bond of N-substituted thymines produced several mono- and di-carboxylic acids, among which N-substituted derivatives of 5,6-dihydrothymine-6-carboxylic acid [5-methyldihydroorotic acid (5-methyl-DHO)] were produced in the highest yield. Similar carboxylation by CO2 radical anions was also observed for thymine dinucleoside monophosphate. The X-ray structure of cis-5,6-dihydro-1-methylthymine-6-carboxylic acid (cis-1,5-dimethyl-DHO) was determined to show a chair conformation in the crystal. CONCLUSIONS: The CO2 radical anion is a nucleophilic radical with rather low reduction potential, thereby possessing a dual reactivity of radical addition preferentially at C6 and one-electron reduction towards thymine-related compounds.

A novel class of antitumor prodrug, 1-(2'-oxopropyl)-5-fluorouracil (OFU001), that releases 5-fluorouracil upon hypoxic irradiation.

We have been developing prodrugs of anticancer agents such as 5-fluorouracil (5-FU) that are activated by irradiation under hypoxic conditions via one-electron reduction. Among them, OFU001 [1-(2'-oxopropyl)-5-fluorouracil] is a prototype radiation-activated prodrug. In this study, we investigated the radiation chemical reactivity and the biological effects of OFU001. This prodrug is presumed to release 5-FU through incorporation of hydrated electrons into the antibonding sigma * orbital of the C(1')-N(1) bond. Hydrated electrons are active species derived from radiolysis of water, but are readily deactivated by O(2) into superoxide anion radicals (O(2).(-)) under conditions of aerobic irradiation. Therefore, 5-FU release occurs highly specifically upon irradiation under hypoxic conditions. OFU001 dissolved in phosphate buffer released 5-FU with a G-value (mol number of molecules that are decomposed or produced by 1 J of absorbed radiation energy) of 1.9 x 10(-7) mol / J following hypoxic irradiation, while the G-value for 5-FU release was 1.0 x 10(-8) mol/J following aerobic irradiation. However, the G-values for decomposition of OFU001 were almost the same, i.e., 3.4 x 10(-7) mol/J following hypoxic irradiation and 2. 5 x 10(-7) mol / J following aerobic irradiation. When hypoxically irradiated (7.5 - 30 Gy) OFU001 was added to murine SCCVII cells for 1 - 24 h, a significant cell-killing effect was observed. The degree of this cytotoxicity was consistent with that of authentic 5-FU at the corresponding concentrations. On the other hand, cytotoxicity was minimal when the cells were treated with aerobically irradiated or unirradiated OFU001. This compound had no radiosensitizing effect against SCCVII cells under either aerobic or hypoxic conditions when the drug was removed immediately after irradiation. Since hypoxia is generally most marked in tumors and irradiation is applied at the tumor site, this concept of prodrug design appears to be potentially useful for selective tumor treatment with minimal adverse effects of anticancer agents.

Fiber-type-specific alphaB-crystallin distribution and its shifts with T(3) and PTU treatments in rat hindlimb muscles.

Changes in alphaB-crystallin content in adult rat soleus and extensor digitorum longus (EDL) were examined after 8 wk of 3,5, 3'-triiodothyronine (T(3)) and propylthiouracil (PTU) treatments. Cellular distributions of alphaB-crystallin expression related to fiber type, and distribution shifts with these treatments were also examined in detail from the gray level of reactivity to specific anti-alphaB-crystallin antibody. alphaB-crystallin content in both soleus and EDL muscles was significantly decreased after T(3), and that in EDL was significantly increased over twofold after PTU treatment. In both control soleus and EDL muscles, the gray level of type I fibers was higher than that of type II fibers. alphaB-crystallin expression among type II subtypes was muscle specific; the order was type I > IIa > IIx > IIb in control EDL muscle and type IIx > or = IIa in soleus muscle. The relation was basically unchanged in both muscles after T(3) treatment and was, in particular, well maintained in EDL muscle. Under hypothyroidism conditions with PTU, the mean alphaB-crystallin levels of type IIa and IIx fibers were significantly lower than levels under control conditions. Thus the relation between fiber type and the expression manner of stress protein alphaB-crystallin is muscle specific and also is well regulated under thyroid hormone, especially in fast EDL muscle.

Vitamin D3 analogue KH1060 combined with TPA synergistically induces mature macrophages in human myeloblastic leukemia ML-1 cells.

A human myeloblastic leukemia cell line, ML-1, was induced to differentiate along the monocytic lineage following exposure to a 1,25-dihydroxyvitamin D3 analogue, 20-epi-22-oxa-24a,26a,27a-tri-homo-1,25-dihydroxyvitam in D3 (KH1060) or 12-O-tetradecanoylphorbol-13-acetate (TPA). The combination of KH1060 and TPA synergistically induced differentiation of ML-1 cells into mature macrophages with multinuclei. Maturation was also observed in other differentiation characteristics such as phagocytic activity, a-naphthyl acetate esterase activity, and expression of surface antigen CD14. Differentiated ML-1 cells showed attenuation of telomerase activity and cessation of proliferating activity based on evaluation of the expression of genes related to cell growth potential. Remarkable synergism was observed in TNF production. Treatment with TPA prior to KH1060 resulted in only slight production of TNF; however, treatment with KH1060 preceding TPA induced a substantial amount of TNF.

Comparison of egg-yolk protein hydrolysate and soyabean protein hydrolysate in terms of nitrogen utilization.

Egg-yolk protein hydrolysate (YPp) is an alternative protein source in formulas for infants with intolerance to cow's milk or soyabean protein, or for patients with intestinal disorders. However, the nutritional value of YPp has never been investigated. YPp was prepared by enzymic hydrolysis of delipidated yolk protein, which led to an average peptide length of 2.6 residues. Three experiments were performed. In Expt 1, we compared the intestinal absorption rate of YPp and soyabean protein hydrolysate (SPp) in rats. YPp and SPp solutions were injected into the duodenum of anaesthetized rats and blood samples were taken from the portal vein at 7, 15, 30, 60, and 120 min. A higher amino acid concentration in the serum of the YPp group demonstrated that YPp was absorbed faster than SPp. In Expt 2, the effects of dietary YPp and SPp on body-weight gain, protein efficiency ratio (PER) and feed efficiency ratio (FER) were determined. At the end of the experiment, body weight had increased in both groups, while PER and FER were significantly higher in rats fed on YPp. In Expt 3, to investigate the effects of dietary YPp and SPp on N metabolism, we determined the biological value and net protein utilization. Yolk protein was the reference protein. Biological value and net protein utilization values were very similar between animals fed on yolk protein and YPp diets, and significantly higher than in rats fed on the SPp diet. The present findings demonstrate that there is no adverse effect of hydrolysis of yolk protein on N utilization, and that the nutritive value of YPp is similar to that of yolk protein and superior to that of SPp.

[Investigation on daily life concerning patients with spinocerebellar degeneration].

Patients with spinocerebellar degeneration show gradual progression of symptoms and decreasing ADL (activities of daily living), resulting in their having many problems concerning daily care. However, there are relatively few patients with such diseases who have participated in those education and consultation services provided by the public health center. To better meet the needs of these patients, we investigated their attitudes and the various parameters affecting their actual daily life. A survey of patients in Sakai-city was conducted via a questionnaire mailed to 100 patients who applied for financial aid for spinocerebellar degeneration in 1996. A total of 74% of the patients responded to the questionnaire. Patients over 50 years old accounted for 77% of the total, Among all responding patients, 14% had their own occupation. The average period of morbidity was 4.8 years. Most (62%) patients were being treated as outpatients. Main symptoms of the patients were associated with speech and trunk movement. About one-fourth of the patients required constant assistance for bathing and transportation. The main person providing care for the patients was the spouse. Main demands for health and welfare services were, "consulting on care and daily life", and "the opportunity for patients to talk with each other." The need for health services is high because such patients have several symptoms. Information acquired through this investigation from patients and their families will be useful for providing better care for patients, with spinocerebellar degeneration.

Hydroxyl radical-induced cross-linking of thymine and lysine: identification of the primary structure and mechanism.

Hydroxyl radical-induced formation of a cross-link of thymine (Thy) and lysine (Lys) in the gamma-radiolysis of N2O-saturated aqueous solution was studied. A Thy-Lys cross-link (I) of the formal structure that OH radical and 4-carbon-centered Lys radical added respectively to C(5) and C(6) positions of Thy was isolated by a preparative HPLC and identified by a FAB-HRMS. The primary cross-link I was dehydrated by treatment with HCl at 120 degrees C to yield the secondary structure (II) possessing a C(5)-C(6) double bond in the Thy moiety: the latter structure II was reported previously (Dizdaroglu, M.; Gajewski, E. Cancer Res. 1989, 49, 3463-3467). A pulse radiolysis study with a redox titration method indicated that 4-carbon centered Lys radical intermediate was of neutral redox reactivity in contrast to reducing reactivity of 5-hydroxy-5,6-dihydrothymin-6-yl radical intermediate. The cross-link I could be formed by a conventional radical recombination mechanism, but not by an ionic recombination mechanism involving a redox reaction between the radical intermediates.

[Association of participation in health checkups with medical service utilization in a rural town].

Association of participation in health checkups in 1983-94 with medical service utilization in 1989-94 was studied. The study subjects consisted of 1,198 males and 1,247 females aged 30 years or over, who were members of the National Health Insurance in a rural town in Kyoto prefecture. One hundred and four males and 95 females died during 1989-94. Cumulative death rates were higher among both males and females with less participation in checkups than among those with higher participation, however, there was no difference seen for medical service utilization. Regarding subjects alive in 1994, both males and females who participated frequently in checking in 1983-88 also visited medical facilities more frequently and had higher medical costs in 1989-94 than those with less frequent participation. Similar analysis for checkups in 1989-94, showed that only females had that tendency. Among participants in checkups in 1989-94, there were smaller numbers of hypertensives, current smokers, and current drinkers than among non-participants, therefore, the participants are thought to have decreased their risk for chronic diseases. In conclusion, frequent participation in health checkups are thought to increase medical utilization even with a lower prevalence or risk of chronic diseases.