Search

Background and Objective: Tomato seeds are edible seeds unconsciously ingested with the fruit. However, there are few reports regarding the constituents and biological activities of tomato seed extract (TSE). Recently, we found that saponins are major constituents of TSE including lycoperoside H. Previous reports have described that several plant-derived saponins improve skin diseases such as wounds and microangiopathy. Therefore, to discover the effect of TSE on the skin condition, we conducted a clinical trial of TSE (Tomato Seed Extract-P) standardized with lycoperoside H when orally ingested. Methods: The study was performed as a randomized, double-blind, placebo-controlled study. TSE (200 mg daily) containing 1 mg of lycoperoside H was used as the active sample. We enrolled 44 Japanese women who have concerns about facial elasticity and relatively low facial skin elasticity. All subjects were randomly allocated into either the active group (n = 22) or the placebo group (n = 22) using a computerized random-number generator. Capsules containing either the active sample or a placebo were administered for 8 weeks between October 12, 2020, and January 16, 2021. Facial elasticity, specifically the R7 value, was evaluated as the primary outcome. The remaining facial R parameters, upper arm R parameters, and other skin parameters including epidermal moisture, trans epidermal water loss, dermal parameters, and advanced glycation end products (AGEs) parameters were measured at 0, 4, and 8 weeks of ingestion. Blood, urine, and body parameters were also evaluated for safety. Results: Forty-three subjects completed the trial, and the per protocol set comprised 21 subjects in the TSE group and 22 subjects in the placebo group. After ingesting TSE for 8 weeks, the R7 value was significantly higher in the TSE group compared to the placebo group. Furthermore, the change in R7 values from the baseline at 4 and 8 weeks were also higher in the TSE group. Among the secondary outcomes, facial elasticity parameters including R2, R5, R1, and R4 at 4 weeks and facial R5, R1, and R4 and upper arm R2 at 8 weeks were higher in the TSE group. In addition, plasma pentosidine significantly decreased in the TSE group after 8 weeks of ingestion. There were no significant differences in moisture, DermaLab® parameters and AGEs parameters except plasma pentosidine. Laboratory tests revealed no abnormalities suggesting adverse effects of TSE. Conclusions: TSE (200 mg/day) standardized with lycoperoside H improved the facial elasticity parameters. Thus, daily ingestion of TSE was suggested to be beneficial for maintaining the facial skin elasticity. However, the relationship between the reduction of pentosidine and skin elasticity by TSE ingestion should be clarified through further studies. Trial Registration: UMIN-CTR: UMIN000041881. Foundation: Oryza Oil & Fat Chemical Co., Ltd.

A high-throughput screening method for the complexation between metal ions and drugs was established by combining solid-phase extraction (SPE) using a nitrilotriacetic acid (NTA) modified silica spin cartridge with subsequent HPLC analysis. First, a test metal ion solution was passed through the NTA cartridge, then a test drug solution diluted in phosphate buffered saline (pH 7.4) was passed through the metal-chelated NTA cartridge. The complexation behavior between the metal and the drug on the NTA cartridge was evaluated by HPLC quantification of the drug in the SPE eluate. Comprehensive analysis of the complexation behavior between 11 different metal ions and 55 drugs showed that Cu2+, Ni2+, Co2+, Zn2+, Cr3+ and Fe3+ formed complexes with 12, 5, 4, 2, 1 and 1 kinds of drugs, respectively. Bromazepam selectively formed complexes with Cu2+, Ni2+ and Co2+.

Self-assembled structures of hexa-tert-butyl-hexa-peri-hexabenzocoronene (HB-HBC) on the Si(111)- 3 × 3 -Ag surface have been investigated by low-energy electron diffraction and low-temperature scanning tunneling microscopy (STM). The HB-HBC molecules are assembled into a 21 × 21 − R ± 10 . 9 ∘ structure with respect to the Si(111)- 1 × 1 surface. Although the periodic structure is unchanged as a function of molecular coverage, we find that the orientations and adsorption sites of HB-HBC within the molecular islands are changed at near a full monolayer. At submonolayer coverages, the tert-butyl groups of HB-HBC are adsorbed on both the Si and Ag trimers of the Si(111)-Ag surface, and only on the Si trimers at a full monolayer. This result should be associated with a balance between substrate-molecule and inter-molecule interactions.

Background and objective: The ingestion of plant-derived glucosylceramides (GlcCer) has been reported to contribute to skin barrier function and hydration of the epidermis. b-sitosterol glucoside (BSG) colocalized with GlcCer in the rice hydrophobic fraction has been shown to increase ceramides in the stratum corneum in vitro . Although clinical studies demonstrated that GlcCer reduced transepidermal water loss (TEWL), the contribution of BSG to epidermal dehydration when applied with GlcCer remains unknown. Therefore, we herein conducted a clinical trial on the effects of a rice-derived mixed fraction of GlcCer and BSG (Oryza Ceramide ® ) on TEWL and other skin parameters. Methods: A randomized, double-blind, placebo-controlled study design was used. Oryza Ceramide ® (type PCD, 40 mg daily) containing 1.2 mg of GlcCer and 40 mg of BSG was used as the active sample. We enrolled 44 healthy Japanese women with epidermal dehydration. All subjects were randomly allocated to an active group (n=22) or placebo group (n=22) using a computerized random number generator. Capsules containing the active sample or placebo were administered for 12 weeks between August and December 2020. Cheek TEWL after 12 weeks was assessed as the primary outcome, and TEWL on a different part of the skin and various skin parameters, including epidermal moisture, pigmentation, pores, and elasticity, were measured before and after 4, 8, and 12 weeks of the intervention. Blood, urine, and body parameters were also examined to evaluate safety. Results: Forty-four subjects completed the trial, and the per protocol set comprised 22 each in the active and placebo groups. Cheek TEWL significantly reduced after the Oryza Ceramide ® intervention for 4 and 12 weeks. Among the secondary outcomes examined, lip moisture (12 weeks) and visible pore number (4 weeks) were improved by Oryza Ceramide ® . Laboratory tests revealed no abnormalities to suggest any adverse effects of Oryza Ceramide ® . Conclusions: Oryza Ceramide ® (40 mg/day) consisting of GlcCer and BSG improved facial TEWL, lip moisture, and visible pores, and these effects may be attributed to increases in epidermal ceramides. The combination of rice GlcCer and BSG appears to be beneficial for improving facial skin conditions. Trial Registration : UMIN-CTR: UMIN000041295 Foundation : The study was funded by Oryza Oil & Fat Chemical Co., Ltd. and Aichi Prefectural Subsidies for Research and Development of Creative Products in 2020. Keywords: rice; glucosylceramide; β -sitosterol glucoside; trans epidermal water loss; pore

Heart failure often presents with prognosis-relevant impaired renal function. To investigate whether the chronic activation of guanylate cyclase-A (GC-A) protects both heart and kidney, we examined the effects of TDT, a neprilysin (NEP)-resistant natriuretic peptide (NP) derivative, on cardiac and renal dysfunction in Dahl salt-sensitive hypertensive (DS) rats. Pretreatment with NEP or NEP inhibitor did not influence GC-A activation by TDT both in vitro and in vivo, resulting in a long-acting profile of TDT compared with native human atrial NP (hANP). The repeated administration of TDT to DS rats suppressed the progress of cardiac hypertrophy, systolic/diastolic dysfunction, and proteinuria in a dose-dependent manner. Compared with vehicle and hANP, salt diet-induced podocyte injury was reduced by TDT, as analyzed by urinary podocalyxin concentration, renal expression of nephrin mRNA, and glomerular expression of desmin protein. Since glomerular TRPC6 plays detrimental roles in podocyte homeostasis, we examined the renal expression of TRPC6 in DS rats and found that salt diet upregulated the expression of TRPC6. Importantly, TRPC6 induction was significantly decreased in TDT-treated rats, compared with vehicle and hANP. Consistently, in primary-culture podocytes from DS rats, TDT inhibited ATP-induced calcium influx, similar to TRPC inhibitor SKF96365. Finally, TDT-mediated protection of podocytes was abolished by protein kinase G inhibitor KT5823. In conclusion, TDT treatment attenuated heart and kidney dysfunction, accompanied by podocyte protection through inhibition of TRPC6. Thus, long-acting NPs could be a new avenue for treatment of heart failure.

Purpose This study was designed to evaluate the efficacy and safety of a compound supplement containing glucosamine (GS), chondroitin sulfate (CS), and five bioactive ingredients for improvement of knee osteoarthritis (OA). Methods Sixteen volunteers aged ≥40 years with knee pain and without ambulant treatment participated in a 6-week randomized, double-blind, placebo-controlled study. They were assigned to the dietary supplement or placebo groups (n = 8, respectively) and ingested six capsules twice daily. The OA symptoms of each subject were determined in pre- and post-treatment periods using a visual analog scale for pain (VAS pain) and four Japanese Knee Osteoarthritis Measure (JKOM) subscales ("joint stiffness," "daily living," "social activities," and "general health condition"). For safety and biomarker assessments, blood and urine samples were tested. Results In the treatment group, the subjective symptoms of VAS pain and three JKOM subscale scores except for "social activities" were significantly improved compared to pre-treatment. Among them, the pattern of change in "joint stiffness" and total JKOM scores showed a significant difference between groups (p = 0.008 and 0.041, respectively). The serum level of interleukin-6, a systemic inflammation biomarker, was significantly decreased in the treatment group after 6 weeks (p = 0.019), whereas the level remained stable in the placebo group (p = 0.690). Diagnostic urine and hematological parameters revealed no serious adverse differences following dietary supplementation over the 6-week study. Conclusion These findings suggest that the compound supplementation of functional food ingredients has potential as an adjunctive and safe therapy for knee OA.

Pyrroloquinoline quinone (PQQ) is a water-soluble quinone compound that has a strong anti-oxidant capacity. A previous study in rats fed a PQQ-depleted diet showed that elevated levels of serum triglyceride (TG) decreased after PQQ supplementation. However, there is only one study reporting the effects of PQQ on serum lipid levels, such as those of TG and cholesterol, in humans. In this study, the effects of PQQ disodium salt (BioPQQ™) on serum TG and cholesterol levels in humans after 6 and 12 wk of treatment at an oral dosage of 20 mg/d were examined. This trial was conducted according to a randomized, placebo-controlled, double-blinded protocol. A total of 29 healthy Japanese adults, ranging from 40 to 57 y old, with normal to moderately high TG levels (110-300 mg/dL) as measured by a recent blood examination, were included in this study. In eleven volunteers out of 29, serum low-density lipoprotein cholesterol (LDL-chol) levels at baseline were high (≥140 mg/dL). After 12 wk, the mean serum TG levels had not changed; however, a marginally significant decrease in the mean LDL-chol (from 136.1 to 127.0 mg/dL) was observed in the PQQ group. In the stratification analysis of the high LDL-chol subgroup (baseline LDL-chol level ≥140 mg/dL), the mean LDL-chol levels decreased significantly from the baseline values in the PQQ group compared to the placebo group. Our study findings suggest that PQQ suppressed the LDL-chol level, which is an important finding, because a high level of this lipid is a risk factor for various lifestyle-related diseases.

Prevotella intermedia is a periodontal pathogen that requires iron for its growth. Although this organism has hemolytic activity, the precise nature of its hemolytic substances and their associated hemolytic actions are yet to be fully determined. In the present study, we identified and characterized several putative hly genes in P. intermedia ATCC25611 which appear to encode hemolysins. Six hly genes (hlyA, B, C, D, E, and hlyI) of P. intermedia were identified by comparing their nucleotide sequences to those of known hly genes of Bacteroides fragilis NCTC9343. The hlyA-E, and hlyI genes were overexpressed individually in the non-hemolytic Escherichia coli strain JW5181 and examined its contribution to the hemolytic activity on sheep blood agar plates. E. coli cells expressing the hlyA and hlyI genes exhibited hemolytic activity under anaerobic conditions. On the other hand, only E. coli cells stably expressing the hlyA gene were able to lyse the red blood cells when cultured under aerobic conditions. In addition, expression of the hlyA and hlyI genes was significantly upregulated in the presence of red blood cells. Furthermore, we found that the growth of P. intermedia was similar in an iron-limited medium supplemented with either red blood cells or heme. Taken together, our results indicate that the hlyA and hlyI genes of P. intermedia encode putative hemolysins that appear to be involved in the lysis of red blood cells, and suggest that these hemolysins might play important roles in the iron-dependent growth of this organism.

We encountered a 60-year-old woman with remarkably elevated thyroid-stimulating hormone (TSH) level as measured by electrochemiluminescent immunoassay (ECLIA), but with no specific symptoms, and with normal levels of free T3 and free T4. We performed the following investigations: polyethylene glycol (PEG) precipitation test, human antimouse IgG antibody (HAMA) interference test, and 3 additional TSH measurements by chemiluminescent immunoassay (CLIA). We then performed 2 gel filtration chromatography (GFC) procedures; one was at pH 7.2, and the other was at pH 3.0. Although the recovery of TSH shown by the PEG precipitation test was 4% which was extremely low, no HAMA interference was observed. Moreover, 3 CLIA instruments also showed various high values. The first GFC showed that the main peak of TSH immunoreactivity by ECLIA was located at a slightly larger molecular weight position than that of IgG. By the second GFC, the sample from the peak fraction of the first GFC showed that the TSH peak disappeared completely at the previous retention time but newly appeared at the same retention time as the TSH monomer. Protein G-Agarose gel removed the majority of the TSH complex. In conclusion, the majority of TSH in her serum was macro-TSH; TSH and anti- TSH IgG autoantibody complex. We should keep the possibility of macro-TSH in mind in cases with unexpectedly high TSH values, especially in autoimmune thyroidal disorders.

For the purpose of marine environmental radioactivity monitoring, stable Cs and 137Cs concentrations were obtained from 214 marbled flounder (Pleuronectes yokohamae) samples collected from off the coast of Ibaraki prefecture in Japan in 2005 and 2006. Prior to the investigation, an analytical method for the determination of Cs in muscle of marbled flounder samples by inductively coupled plasma mass spectrometry (ICP-MS) was seriously considered from the view point of sample preparation (wet way) to a Cs measurement. Since major problems in determining Cs by the ICP-MS is the interference of matrix materials and molecular ions on Cs peak area (m/z 133), a known amount of In (m/z 115) was added to the sample solutions as an internal standard. To check the accuracy of the determination, duplication analysis and standard reference materials were used. Analytical results of Cs in muscles samples by ICP-MS agreed well with duplicated samples, and standard reference materials. The standard length and body weight were obtained from a total of 214 flounder samples. The stable Cs, stable K and 137Cs concentrations in muscle were determined by ICP-MS, flame photometry and radiochemical analysis, respectively. The mean standard length correlates closely with the mean body weight. The stable Cs concentration was in proportion to the body weight, also, the 137Cs concentration correlated closely with the stable Cs concentration in muscle. On the other hand, a stable K concentration was found to be uniform, and unrelated with 137Cs and stable Cs concentrations. The specific activities, the ratio of 137Cs/stable Cs in muscle, was found to be 5.5∼6.7 with an average of 6.2±0.35 (SD) among five different size groups of the standard length of marbled flounder samples. From these results, ICP-MS provided useful tools for the determination of a trace amount of Cs in muscle samples, combined with wet way pretreatment techniques. In addition, marbled flounder proved to be a useful species for comparing the radioactivity of fishes in coastal seas around Japan with each other in the environmental radioactivity monitoring program.

We have characterized the structures of ganglioside (GD3) micelle and GD3/cholesterol mixtures, and have determined the permeability of water across a membrane of GD3/cholesterol/phospholipid uni-lamellar vesicle. We have found that monovalent Na+ and K+ cations affect the permeability of water differently. We have found also that the permeability of water is greatly enhanced by the K+ ion, suggesting the function of the GD3/cholesterol rich clusters on neuronal excitations by K+. The complimentary use of small-angle X-ray and neutron scattering techniques is useful to determine the permeability of water across a vesicle membrane.

Matrix metalloproteinases (MMPs) play a crucial role in the process of cancer invasion and metastasis. Previous findings suggested that epigallocatechin gallate (EGCG), a main flavanol of green tea, caused decreased levels of MMP-2 and MMP-9 activities to be secreted into culture medium. To obtain further information on EGCG-mediated regulation of these MMPs, the effects of EGCG on enzyme activity, mRNA expression, and mitogen-activated protein kinase (MAPK) activities in human fibrosarcoma HT1080 cells were examined. EGCG was confirmed to suppress the gelatin-degrading activities due to MMP-2 and MMP-9 in the culture medium. This suppression of enzyme activities by EGCG was consistent with the decreased levels of MMP-2 and MMP-9 mRNAs. EGCG-mediated suppression was also observed for MT1-MMP mRNA. EGCG-mediated suppression of the level of MMP-9 transcript was correlated with its suppression of MMP-9 promoter activity. EGCG inhibited the phosphorylation of extracellular signal-regulated kinases 1 and 2 (ERK1/2), which are the members of an MAPK family necessary for MMP-9 up-regulation. EGCG also suppressed p38 MAPK activity but gave no effects on stress-activated protein kinase/c-Jun N-terminal kinase activity. These findings suggest that suppression of ERK phosphorylation by EGCG is involved in the inhibition of expression for MMP-2 and MMP-9 mRNAs, leading to the reduction of their enzyme activities of the cancer cells. Methyl derivatives, epigallocatechin-3-O-(3-O-methyl) gallate and epigallocatechin-3-O-(4-O-methyl) gallate, exhibited effects similar to, but weaker than, those of EGCG, suggesting the important role of an unsubstituted triphenolic ester structure in these activities.

1. To investigate the pharmacological effects of T-1095, this novel derivative of phlorizin was administered to GK rats for 8 weeks. T-1095 treatment significantly lowered plasma glucose and glycosylated haemoglobin (HbA1c) levels, but did not significantly affect bodyweight. 2. T-1095 treatment did not affect 3.3 mmol/L glucose-induced insulin secretion in the isolated perfused pancreas of GK rats. 3. The peak insulin release in T-1095-treated GK rats was significantly higher during the first phase than in untreated GK rats (3-4 min after beginning 16.7 mmol/L glucose perfusion). The total amount of insulin secreted during the first phase in T-1095-treated GK rats was significantly higher than in untreated GK rats (35.3 +/- 1.4 vs. 27.3 +/- 2.5 ng in T-1095-treated compared with untreated rats, respectively). 4. During the second phase, insulin release in T-1095-treated GK rats was somewhat higher than in untreated GK rats (7-30 min after beginning 16.7 mmol/L glucose perfusion). The total amount of insulin secreted during the second phase in T-1095-treated GK rats was significantly higher than in untreated GK rats (88.2 +/- 6.1 vs. 68.1 +/- 5.7 ng, respectively). 5. The total amount of insulin secreted during perfusion in T-1095-treated GK rats was significantly higher than in untreated GK rats (123.5 +/- 7.3 vs. 95.4 +/- 7.7 ng, respectively). 6. These data show that the metabolic indices, plasma glucose and HbA1c levels and insulin secretion are significantly improved by T-1095 treatment in GK rats, which are spontaneously diabetic rats, suggesting its usefulness as a novel oral therapeutic antidiabetic agent.

Background: The seaberry is a yellow berry cultivated in China, Northern Europe, and West Asian countries. Numerous biological activities of seaberries have been reported, and we recently found that ursolic acid and a flavonoid in seaberry extract (SBE) suppressed the contraction of bladder muscle specimens and collagen gel containing bladder smooth muscle cells. However, the influence of SBE on urinary problems of Japanese adults has not been investigated. Therefore, we conducted a clinical trial of SBE supplementation in Japanese subjects with mild urinary dysfunction. Methods: We examined the effect of SBE (200 or 400 mg daily) containing ursolic acid (0.18 or 0.36 mg) in a randomized double-blind placebo-controlled study. Capsules containing SBE or placebo were administered for 8 weeks to Japanese men and women with mild urinary dysfunction. After 4 and 8 weeks of treatment, urinary symptoms were evaluated by using the King’s Health Questionnaire (KHQ) and the overactive bladder syndrome score (OABSS). Results: There was significant improvement of the KHQ emotion domain score after intake of 400 mg/day of SBE for 8 weeks compared with placebo. The answers to “Does your bladder problem make you depressed?” and “Does your bladder problem make you feel bad about yourself?” were significantly improved by SBE (400 mg/day) at 8 weeks. In the OABSS, the item “How often do you have a sudden desire to urinate, which is difficult to defer?” was significantly improved by SBE compared with placebo. Laboratory tests did not reveal any abnormalities suggesting adverse effects SBE. Conclusions: Intake of SBE (400 mg/day for 8 weeks) improved several emotional parameters related to urinary dysfunction. SBE may be useful for reducing moderate urinary symptoms. Keywords : King’s health questionnaire; overactive bladder syndrome score; seaberry; ursolic acid; overactive bladder; urination; micturition

This is a report on a capillary electrophoretic approach for the quantitation of the counter ions of drugs that are insoluble or slightly soluble in water. Using a nonaqueous run buffer, the sample solvents were not restricted, which is not the case with conventional aqueous run buffer systems. The separation selectivity could be manipulated using mixed solvents such as methanol and dimethylformamide as run buffers. Detection was carried out by indirect UV detection with phthalate. The linearity and the repeatability of the peak area of chloride was satisfactory. This approach was applied to the drug substance, azelastine hydrochloride, which is slightly soluble in water, and provided a value of chloride content consistent with the theoretical value.

By means of the inverse contrast variation method in small-angle neutron scattering, we have studied the structure of a small unilamellar vesicle (SUV) composed of ganglioside, cholesterol and dipalmitoyl-phosphocholine. The SUV treated has a similar lipid composition as in a plasma membrane with microdomains, so-called rafts. The present results indicate an asymmetric distribution of lipid components within the bilayer of the vesicle, that is, a predominant distribution of ganglioside and cholesterol at the outer leaflet of the vesicle bilayer. The deviation from the linearity in a pseudo-Stuhrmannplot strongly suggests the presence of a large heterogeneity of lipid composition in a bilayer, namely a clustering of ganglioside and cholesterol molecules. This deviation is enhanced by temperature elevation, meaning that ganglioside–cholesterol clusters become larger with holding liquid-ordered (Lo) phase.

Synthesis of the tetracyclic core ( 19 ) of manzamine A ( 1 ) was achieved via Diels-Alder reaction of the dihydropyridinones ( 5 , 6 ). Conversion of the two D-A products ( 7 , 8 ) to the key tricyclic ketone ( 10 ) was conducted through a conventional pathway (Scheme III) as well as a new pathway developed (Scheme IV). For effective construction of the required azocine ring systems, model studies were carried out to find intramolecular amide formation by pentafluorophenyl ester and DPPA methods, which were successfully applied to the real substrate to furnish the titled core structure ( 19 ).

Effects of S-23142 on protoporphyrin IX (Proto IX) biosynthesis in chloroplasts isolated from Spinacia oleracea L. were examined using reverse phase HPLC with fluorescence monitoring. The synthesis of Proto IX was inhibited to a level of 50% by 10-9 ᴍ of S-23142 in this system. The effects of S-23142 was also tested in chloroplasts isolated from two types of photomixotrophic tobacco cells, wild type and S-23142 tolerant cells. The biosynthesis of both the wild type cells and YZI-1 S cells were inhibited at 50% by 10-9 ᴍ and 10-7 ᴍ of S-23142, respectively. It is, therefore evident that the mutation in the tolerant cell is associated with the Protox. To investigate the localization of Proto IX biosynthesis and the target site of S-23142, spinach chloroplasts were osmotically broken and separated into stroma and membrane (thylakoid and envelope) fractions. A very active Proto IX synthesis from ALA was found in the stromal fraction, while no activity of Proto IX synthesis was observed in the membrane fraction. These results suggest that most Proto IX synthetic activity and a target-site of S-23142 exist in the stromal fraction.

Heating the aqueous solutions of asparagine-containing dipeptides in which asparagine locates at their C-terminal gave sequential polypeptides having the molecular weight up to 3000–5000.

In previous studies, we showed that pretreatment of rat FRTL-5 thyroid cells with TSH, or other agents that increased intracellular cAMP, markedly potentiated DNA synthesis in response to insulin-like growth factor-I (IGF-I). In addition, we found that TSH pretreatment caused an increase in tyrosine phosphorylation of intracellular proteins including an unidentified 125-kDa protein that was well correlated with the TSH-potentiating effect on DNA synthesis induced by IGF-I. These results suggested that cAMP amplified IGF-I-dependent signals for cell growth through changes of cAMP-dependent tyrosine phosphorylation. The present studies were undertaken to determine how tyrosine kinase activation followed by an increase in tyrosine phosphorylation is required for cAMP-dependent potentiation of DNA synthesis induced by IGF-I in this cell line. First of all, we measured tyrosine kinase or protein-tyrosine phosphatase activities in the cell lysates by the in vitro assay. Chronic treatment with TSH or (Bu)2-cAMP stimulated tyrosine kinase activity in the particulate fraction and protein-tyrosine phosphatase activity in the soluble fraction, suggesting that tyrosine kinase plays more important roles for a cAMP-dependent increase in tyrosine phosphorylation of intracellular proteins. The increased tyrosine kinase activity was sensitive to genistein, a potent tyrosine kinase inhibitor. Genistein abolished both the cAMP-dependent increase in tyrosine phosphorylation of the 125-kDa protein and the enhanced DNA synthesis induced by IGF-I in a similar concentration-dependent manner. The only tyrosine-phosphorylated protein associated with the p85 regulatory subunit of phosphatidylinositol (PI) 3-kinase in response to cAMP was 125 kDa. In addition, we found that PI 3-kinase activity bound to p85 subunit significantly increased after (Bu)2cAMP treatment. These results suggested that cAMP stimulates PI 3-kinase through tyrosine phosphorylation of the 125-kDa protein. We then measured DNA synthesis in cells pretreated for 24 h with TSH or (Bu)2cAMP in the absence or presence of LY294002, a PI 3-kinase inhibitor, followed by treatment with IGF-I for 24 h. Presence of LY294002 during TSH or (Bu)2cAMP pretreatment completely abolished cAMP-dependent potentiation of DNA synthesis induced by IGF-I. These results suggest that in FRTL-5 cells cAMP activates genistein-sensitive tyrosine kinases that in turn activate PI 3-kinase activity. These mechanisms appear to be necessary for cAMP-dependent potentiation of the DNA synthesis induced by IGF-I.

A pot experiment was conducted with an Andosol to investigate the changes of phosphorus(P) compounds in the rhizosphere and hyphesphere soil (Organic P, Ca-P, Al-P, Fe-P). To distinguish between the effect of the arbuscular mycorrhizal root system and only of mycorrhizal hyphae, “a double pot system” (separation of a root and hyphae compartment by a nylon net) has been used. Ca-P and Al-P fraction around the mycorrhizal roots and mycorrhizal hyphae of both plant species, groundnut and pigeonpea significantly decreased under non-sterilized condition after 100 days of cultivation during the summer. This indicates that Ca-P and Al-P fractions in the Andosol with high accumulated P were quite mobile and available for these plants.

Estrogen deficiency in postmenopausal women is a major contributing factor to bone loss occurring after menopause. Although serum estradiol levels decrease conspicuously, a considerable amount of estrone still exists in women after natural menopause. Since most of the estrone is converted from androgens in adipose tissues, we attempted to seek interrelationship among serum estrone levels, fat mass and bone mineral density (BMD) in postmenopausal women. Thirty women who underwent natural menopause at the age between 45 and 53 were examined for BMD of head, arms, trunk and legs, as well as total body fat and total lean body mass using a Lunar DPX, a dual energy X-ray absorptiometer. Serum estrone concentration in each subject was measured by RIA. BMD of all sites of bones examined showed a positive correlation with relative fat mass (total body fat/body weight). Serum estrone levels were positively correlated with BMD of arms, trunk and legs, as well as relative fat mass. Multiple regression analysis further demonstrated that relative fat mass and serum estrone levels were significantly related to BMD of both arms and trunk, and BMD of trunk, respectively. These results suggest that estrone produced in adipose tissues may have, if only in part, a role in modulating the loss of bone mass occurring after menopause.

L. leichmanniiを用いた牛乳・調製粉乳中のビタミンB12の測定条件につき検討し,以下の結果を得た. (1) 接種用培地の前培養時間は5時間,接種菌液濃度はOD520=0.01,本培養時間は25時間,波長520nmでの測定が最適と思われた.測定時間は従来の32～44時間から30時間に短縮された. (2) 保存菌株の継代培養は毎日実施する必要はなく,1カ月間隔でも活性の低下は認められなかった.416日本食品工業学会誌 第36巻 第5号 1989年5月(60) (3) 新試験法により添加回収実験を試みたところ,良好な結果を得た. (4) 日内・日間分析精度を求めたところ従来法と比較して小さい値を得た.

It is considered that there were several possible pathways in which polypeptides were produced on premitive earth. Of these pathways, (i) condensation of solid amino acids on heating, (2) condensation of amino acids in aqueous media over condensing agents or (3) on heating are known. The first one has been studied in detail(Fox et al., 1960). The second one has not given longer peptides than 5 residues(Hulshof et al., 1976). However, there are a few reports related to the third one in which much larger peptides were obtained from aspargine with the molecular weight more than c.a. 3000(Kovacs et al., 1961.; Harada et al., 1978). In this paper, we wish to describe the role of aspargine and glutamine having 8 or y primary amide for polypeptide formation in the hypothetical premitive ocean. We carried out the heating of aqueous solution contaning aspargine and other amino acids(glycine, alanine, and valine) or peptides which have asparagine or glutamine residue in the C-terminal. Heatin @ reaction of aqueous solution of as~arqine and some other amino acids. Five mmol L-aspargine monohydrate(750mg) and other amino acids[0.5-2.5 mmol(Gly, Ala, Val)]were dissolved in 5 ml H20 , and the solutions sealed in vacuo in a glass tube were heated at 120°C for 2-200 hr. After the reactions, the reaction solution was analyzed by amino acid analyzer and remaining sample was loaded into a Sephadex G-15 column. The elution was carried out with 0.5 M-acetic acid to give white amorophous powder. The 32 hours' reaction mixtures gave muximum isolated yield(>90mg). The IR of this compound showed typical acidic polypeptide absorption(1710, 1650, 1540 cm-l). Averaged molecular weight of this peptide was estimated to be 1500-3000 by gel filtration. Molar composition of the amino acids contained in the copolyamino acids were almost pallarel to the first composition of the reaction mixtures, when the molar ratio of other amino acid to L-aspargine was lower than 50 %. Heating reactions of amino acid solutions without aspargine