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Biomedical subjects

A Yagi

Publications and source records attributed to A Yagi.

At least 55 records · Page 3Linked to original sources
PubMed1996

Antifungal activity from Alpinia galanga and the competition for incorporation of unsaturated fatty acids in cell growth.

An antimicrobial diterpene was isolated from Alpinia galanga in the screening for potentiators of phytochemical antibiotic action. The structure was elucidated by spectral data and identified as (E)-8 beta, 17-epoxylabd-12-ene-15, 16-dial (1). Diterpene 1 synergistically enhanced the antifungal activity of quercetin and chalcone against Candida albicans. Antifungal activity of 1 was reversed by unsaturated fatty acids. Protoplasts of C. albicans were lysed by 1. These results suggest that antifungal activity of 1 is due to a change of membrane permeability arising from membrane lipid alternation.

Antifungal Agents↗
PubMed1996

Instability of bovine insulin in poly(lactide-co-glycolide) (PLGA) microspheres.

Biodegradable poly(lactide-co-glycolide) (PLGA; 50/50) microspheres containing bovine insulin as a model protein was prepared by an oil-in-oil (o/o) emulsion solvent evaporation process. When aluminum tristearate (0.15% (w/v)) was employed as a dispersing agent, the loading efficiency of insulin was almost 100% and the yield was over 80%. The average diameter of the PLGA microspheres always ranged between 100 and 200 microns. Morphology study using a scanning electron micrograph showed smooth, spherical, fairly monodispersed PLGA microspheres containing insulin. In relation to release profile, the very low release rate of insulin was demonstrated (only 1% of insulin released after 7 d release test in pH 7.4 Tris buffer) for the PLGA microspheres. Nevertheless, the degradation of bovine insulin in PLGA microspheres was confirmed by high performance liquid chromatography. This degradation seemed to be caused by an acidic condition caused by poly(lactide-co-glycolide) polymer.

Animals↗
PubMed1996

Cis- and trans-linalool 3,7-oxides and methyl salicylate glycosides and (Z)-3-hexenyl beta-D-glucopyranoside as aroma precursors from tea leaves for oolong tea.

Two new alcoholic aroma precursors, cis- and trans-linalool, 3,7-oxides 6-O-beta-D-apiofuranosyl-beta-D-glucopyranosides (1 and 2), as well as two already known compounds, (Z)-3-hexenyl beta-D-glucopyranoside (3) and methyl salicylate 6-O-beta-D-xylopyranosyl-beta-D-glucopyranoside (beta-primeveroside: 4), and another new monoterpendiol glycoside, 8-hydroxygeranyl beta-primeveroside (5) have recently been isolated as aroma precursors in tea leaves (Camellia sinensis var. sinensis cv. Maoxie) ready for oolong tea processing.

Carbohydrate Sequence↗
PubMed1996

Neonatal estrogenization of the male mouse results in urethral dysfunction.

PURPOSE: Analysis of voiding pattern, urodynamic measurements and immunohistochemical methods were performed in order to evaluate the effects of neonatal estrogenization on voiding functions of adult male mice. MATERIALS AND METHODS: Metabolic cages were used for recording the voiding volumes and frequencies. Bladder pressure and mean flow during voiding were measured in transvesical cystometry. Location of estrogen receptors and organization of smooth muscles in lower urinary track were demonstrated using immunohistochemical staining. RESULTS: Neonatally estrogenized (neoDES) male mice had lower voided urine volumes (the average voided urine volume and average of the three largest volumes) and higher voiding frequencies than control mice. In transvesical cystometry, the maximum bladder pressure during the high-frequency oscillation phase of voiding was significantly elevated. The average urinary flow rate was decreased. CONCLUSIONS: Urodynamically, these findings are consistent with the concept that neonatally estrogenized mice have infravesical obstruction. The predominance of estrogen receptors in the periurethral region and changes in urethral smooth muscle cells immunocytochemically stained with alpha-actin-antibody support the concept of urethral wall musculature as a target of estrogen action.

Age Factors↗
PubMed1995

Actin, alpha-actinin, and spectrin with specific associations with the postacrosomal and acrosomal domains of bovine spermatozoa.

BACKGROUND: Characteristic membrane changes in spermatozoa culminating in acrosome reaction and sperm-egg fusion, and suspected involvement of actin-containing cytoskeleton in membrane changes in general, prompted us to investigate subcellular distribution of actin and actin-binding proteins in bovine spermatozoa subjected to various extractions which sequentially denude the sperm investments. METHODS: Spermatozoa were treated with either 1% SDS, 0.1% Triton X-100, 0.1% Hyamine, or 1 M MgCl2 or were sonicated. Immunostaining of actin, alpha-actinin, spectrin, and acrosin as well as electron microscopic analysis of extracted spermatozoa were carried out. RESULTS: Extractions caused evagination of the acrosomal lamina which retained focal contacts with the inner acrosomal membrane. Extractions further revealed lateral prongs at the anterior border of the postacrosomal sheath. Labeling for alpha-actinin and spectrin was localized in the acrosin-positive acrosomal lamina, neck, and principal piece, the latter containing also relatively extraction-resistant oligomeric or polymerized actin. In the postacrosomal area, actin was accumulated in the extraction-resistant posterior ring structure and anteriorly at the sites apparently related to the lateral prongs. Notably, spectrin reactivity was enhanced by MgCl2 in head, neck, and principal piece, and sonication abolished cytoskeletal immuno-reactivity in the head. CONCLUSIONS: Destabilization of membranes with selected extractions induces changes in the acrosomal lamina mimicking acrosomal vesicle formation. The lateral prongs and posterior ring structure, respectively, may serve as anterior and posterior anchors for the extraction-resistant post-acrosomal sheath. The lateral prongs may also be a merger zone for actin, alpha-actinin, and spectrin with important implication on sperm function. The latter two proteins may be involved in acrosomal vesicle formation. It is apparent that extractions have a significant effect on the detectability of sperm cytoskeletal elements.

Acrosome↗
PubMed1995

Inhibition of lipid peroxidation and superoxide generation by diterpenoids from Rosmarinus officinalis.

Four diterpenoids, carnosic acid (1), carnosol (2), rosmanol (3), and epirosmanol (4), were isolated as antioxidative agents from the leaves of Rosmarinus officinalis by bioassay-directed fractionation. These diterpenoids inhibited superoxide anion production in the xanthine/xanthine oxidase system. Mitochondrial and microsomal lipid peroxidation induced by NADH or NADPH oxidation were also completely inhibited by these diterpenes at the concentration of 3-30 microM. Furthermore, carnosic acid protected red cells against oxidative hemolysis. These phenolic diterpenes were shown to be effective to protect biological systems against oxidative stresses.

Diterpenes↗
PubMed1995

Ordered structure of the crystallized bovine 20S proteasome.

Eukaryotic proteasomes are multicatalytic proteinase complexes with a molecular weight of 750 kDa, containing, respectively, two copies of a hetero-heptamer of alpha-type subunits and one of beta-type subunits, (alpha 1-7 beta 1-7)2. Proteasome was purified from bovine liver and crystallized into a hexagonal system with cell dimensions of a = b = 121.83(2) A, c = 930.68(6) A. A cylindrical particle size of 122 A diameter and 155 A height was determined from the molecular packing in a unit cell. The crystal gave diffraction spots up to at least 4.4 A resolution, which was the minimum spacing of the camera used. The overall temperature factor of the enzyme was estimated to be in the range of 36.2 to 25.8 A2. These results imply that the enzyme complex has a unique ordered structure comprising multisubunits with two types of hetero-heptamer. This ordered structure may facilitate highly organized cooperation of individual functions of subunits within the enzyme complex.

Amino Acid Sequence↗
PubMed1995

Expression of mitochondrial heat shock protein 60 in distinct cell types and defined stages of rat seminiferous epithelium.

Changes in the level of the gene transcript of heat shock protein (hsp)60, a mitochondrial chaperonin, during the cycle of rat seminiferous epithelium and its cellular localization were studied. The seminiferous epithelium showed a cell type-specific expression of hsp60. Immunostaining of adult rat testis revealed localization in Sertoli and Leydig cells. In germ cells, mitochondria of spermatogonia and early primary spermatocytes were immunoreactive for hsp60. Mitochondria of all other germ cell types were completely negative for hsp60. Stage-specific expression of hsp60 was determined from pooled segments of stage-specific microdissected tubules by a combination of Western blotting and polymerase chain reaction (PCR). High concentrations of hsp60 were found in stages I-V and IX-XIV, and low levels were detected in the other stages, i.e., VI-VIII. In stages with high hsp60 expression, spermatogonia divide mitotically, whereas in stages lacking mitosis, the hsp60 level was much weaker. In seminiferous epithelium, two different types of mitochondria are present. Therefore, immunoelectron microscopy was used to differentiate these two morphologically distinct types of mitochondria. The crista type of mitochondria (e.g., in Sertoli cells and spermatogonia) reacted with the antibody against hsp60, whereas hsp60 was negative in so-called "condensed"-type mitochondria found in midpachytene spermatocytes and more advanced germ cells. It could be shown for the first time that expression of the hsp60 gene is regulated during the cycle of the seminiferous epithelium. The results indicate that the gene product is primarily needed during the initial steps of spermatogenesis in which most of the cell divisions occur, while its expression during the differentiation of spermatids and sperm is obviously not necessary. The presence of hsp60 in stages with mitotic activity suggests a very active mitochondrial protein import and protein assembly machinery that generates further mitochondria for the dividing cells.

Animals↗
PubMed1995

Glycosidic aroma precursors of 2-phenylethyl and benzyl alcohols from Jasminum sambac flowers.

Benzyl 6-O-beta-D-xylopyranosyl-beta-D-glucopyranoside (beta-primeveroside) (1), 2-phenylethyl beta-primeveroside (2), and 2-phenylethyl 6-O-alpha-L-rhamnopyranosyl-beta-D-glucopyranoside (beta-rutinoside) (3) were isolated as aroma precursors of benzyl alcohol and 2-phenylethanol from flower buds of Jasminum sambac Ait. The isolation was guided by an enzymatic hydrolysis and GC and GC-MS analyses.

Benzyl Alcohol↗
PubMed1995

Differential distribution of laminin chains in the development and sex differentiation of mouse internal genitalia.

The distribution of laminin chains and basement membranes (BMs) in the ontogenesis and sex differentiation of male and female mouse gonads and mesonephros was studied by conventional and immunocytochemical light and electron microscopy. The alpha 1 (synonymous to A) chain was recognized with MAbs against fragment E3, and three chains of laminin with PAbs raised against EHS-laminin. BMs, which formed around the mesonephric duct, the mesonephric tubules, and the paramesonephric duct, contained the laminin alpha 1 chain. The alpha 1 chain appeared with epithelial differentiation in the developing gonads in both sexes. The alpha 1 chain was first evident around the embryonic gonadal cords and remained, after development, in the BMs of the testicular cords and ovarian follicles. The laminin alpha 1 chain was also detected in BMs of the myoid cells around the epithelial rete cords, and transiently in the surface epithelium and in the corpus luteum. Laminin beta-gamma chains were found in many locations where the alpha 1 chain was not detected. These included the mesenchyme of the early mesonephros, the BMs of blood vessels and surface epithelium in the differentiated testis and ovary, between the theca cells in the ovary, and in some corpora lutea. The morphological differentiation of the BMs of the embryonic testicular cords proceeded rapidly. In contrast, the BM of the ovarian cords remained relatively poorly differentiated during the prenatal phases, and developed concomitantly with the differentiation of the follicles. The results show that BMs in the differentiating internal genitalia are heterogeneous with respect to their laminin chains, and suggest that all known laminin chains must be analyzed in the differentiation of gonadal epithelia for a complete role of the BMs in gonadal sex differentiation.

Animals↗
PubMed1994

Extractions reveal specific argentophilic proteins in rat and bull sperm heads.

BACKGROUND: Silver-stainability (argentophilia) of cytoplasmic structures occurring in spermatids have been localized into the organizing perinuclear theca, but the biochemical nature and structural associations of these proteins with the cytoskeletal and membranous elements are unresolved and, therefore, were the aim of the present study. METHODS: Light and electron microscopic analysis of the silver-stainability in the rat spermatids and spermatozoa was carried out in the intact testis tissue and epididymal spermatozoa and after their chemical and mechanical extraction. Correlation of argentophilia with specific proteins of rat and bovine spermatids and spermatozoa was investigated using a recently developed technique for silver nitrate staining of proteins on nitrocellulose. RESULTS: Sequential formation of the silver-stainable domains seemed to proceed from the argentophilic acrosomal ring. Various extractions indicated that argentophilia in the spermatids and spermatozoa was mainly associated with the perinuclear theca and to some extent to the plasma membrane. Hyamine-soluble extract from spermatozoa of rat and bull revealed only a single argentophilic protein of 130 kDa. Hyamine and SDS-soluble extracts of rat testis tissue contained an additional group of argentophilic polypeptides of lower molecular weight (115, 94, 36, 23, and 21 kDa). CONCLUSIONS: Reduction in the number of argentophilic proteins appears to be involved in a series of changes in the cyto-architecture of developing spermatids. Tentative cytoskeletal nature of argentophilic proteins remains to be identified. Nevertheless, they may have important physical relations with the higher-order organization of the sperm head cytoskeleton and overlying membranes.

Animals↗
PubMed1994

Sertoli cells in testes containing or lacking germ cells: a comparative study of paracrine effects using the W (c-kit) gene mutant mouse model.

BACKGROUND: Paracrine effects of germ cells on Sertoli cell structure were examined in a mouse model with the W locus (dominant white spotting) mutation in which animals with the W/Wv genotype (referred to as mutants) lack virtually all germ cells. RESULTS: Morphometric determination of Sertoli cell parameters in mutant and control (+/+) animals showed that although the testes of mutant animals were about eight times smaller than controls, the numbers of Sertoli cells in the two groups did not differ. Sertoli cell volume, Sertoli cell cytoplasmic and nuclear volumes, and Sertoli cell surface area in mutant animals were significantly smaller than in control animals. Organelle volumes and surface areas, expressed per cell, did not differ significantly in the two groups with one exception: the volume and surface area of smooth endoplasmic reticulum was significantly reduced in mutant animals. Plasma testosterone levels and tissue testosterone levels/testis were normal, indicating that the effects observed in the mutant animal were not a consequence of androgen insufficiency. Plasma FSH was elevated, probably as a consequence of germ cell depletion, and was thought not to affect Sertoli cell parameters observed. CONCLUSIONS: The data suggest that paracrine interactions with germ cells do affect Sertoli cells by modifying the amount of smooth endoplasmic reticulum. These data focus attention on the function of this abundant Sertoli cell organelle in promoting spermatogenesis.

Animals↗
PubMed1994

Immunocytochemical detection of actin and 53 kDa polypeptide in the epididymal spermatozoa of rat and mouse.

BACKGROUND: Presence of immunocytochemically detectable actin in the rat and mouse sperm head has been enigmatic for years. In this study, we demonstrate actin in the perinuclear theca and show that the detection of actin epitopes in the rat and mouse epididymal spermatozoa can effectively be enhanced by pre-extraction of sperm cells with SDS. METHODS: The study with one monoclonal and one polyclonal anti-actin antibody was carried out at conventional and confocal fluorescence and electron microscope level, and by immunoblotting of proteins isolated from the head and tail fractions. RESULTS: In the head of the control methanol-acetone fixed rat spermatozoa, the polyclonal antibody gave a stronger immunostaining in the postacrosomal area and in the perforatorium than the monoclonal antibody. In the mouse sperm head, the monoclonal antibody labeled the ventral edge of the postacrosomal area and slightly the perforatorium, whereas the polyclonal antibody stained the entire perinuclear space. In the SDS-extracted spermatozoa, an intense postacrosomal and perforatorial labeling was obtained with both antibodies but, in particular in the rat spermatozoa, the middle lateral portion of the postacrosomal segment remained unlabeled. Sonication seemed to cause structural modifications which specifically impeded staining with the monoclonal antibody. Both antibodies detected actin in the basal plate and the monoclonal antibody in the neck. Amorphous matrix of the connecting piece showed immunogold labeling. In the tail, the monoclonal antibody recognized actin and a relatively basic 53 kDa polypeptide, whereas the polyclonal antibody reacted with several protein bands. SDS-soluble actin of the tail was addressed to the midpiece and the SDS-insoluble 53 kDa protein profoundly to the outer dense fibers of the principal piece. CONCLUSIONS: Intense labeling of actin in the SDS-extracted rat and mouse spermatozoa was presumably due to the generated demasking of actin epitopes embedded in the perinuclear cytoplasm. The results are important in confirming that actin in the rat and mouse sperm head is not lost during spermiogenesis but apparently contributes to the three-dimensional packing of the mature perinuclear cytoplasm. This study further demonstrates the importance of the methods used in sample preparation and advantages of confocal microscopy when attempting to detect cytoskeletal proteins which, as in spermatozoa, may occur in small quantities.

Actins↗
PubMed1994

Skin permeability of various drugs with different lipophilicity.

The in vitro and in vivo skin permeability of 16 drugs with a wide span of lipophilicity (log P ranging from -0.95 to 4.40) was evaluated with an ethanol/panasate 800 (tricaprylin) (40/60) system as a lipophilic vehicle. The ethanol/panasate 800 (40/60) binary vehicle remarkably improved the in vitro skin permeability of the drugs across excised hairless mouse skin compared with ethanol or panasate 800 as single vehicles. The in vivo skin permeability of the large majority of the drugs across abdominal rat skin also showed high permeation rates and short lag times. The relationship between lipophilicity and skin permeability of the drugs from the ethanol/panasate 800 (40/60) binary vehicle indicated parabolic shapes with their peaks at much greater hydrophilic range (log P: -0.88 for in vitro, -0.83 for in vivo) compared with other past references (log P: 2-3). The results suggest that the lipophilicity of a drug is a main factor for prediction of the skin permeability of the drug and that the ethanol/panasate 800 (40/60) lipophilic binary vehicle would be a good candidate as a vehicle for future clinical application of hydrophilic drugs.

Animals↗
PubMed1994

Flavonoids in Rosmarinus officinalis leaves.

Three new flavonoid glucuronides, luteolin 3'-O-beta-D-glucuronide, luteolin 3'-O-(4"-O-acetyl)-beta-D-glucuronide, and luteolin 3'-O-(3"-O-acetyl)-beta-D-glucuronide, together with hesperidin, were isolated from 50% aqueous MeOH extract of the leaves of rosemary. The structures were established by chemical and spectroscopic methods. Their antioxidant activities were evaluated by a ferric thiocyanate method with hesperidin showing the greatest activity.

Antioxidants↗
PubMed1994

Effect of a peptide from Panax ginseng on the proliferation of baby hamster kidney-21 cells.

An alkaline fraction separated by ion exchange chromatographies from the water extract of Panax ginseng root stimulated the proliferation of baby hamster kidney-21 cells. Separation of the alkaline fraction by MCI-gel CHP 20P column chromatography followed by dialysis provided an active material. By a reversed-phase HPLC the active material was separated into six fractions, and an active colorless compound 1 was obtained from fraction 2 in a pure state. Compound 1 was composed of the following amino acids; Gly, Arg, Glu, Val in a ratio of 1:1:1:1, and caused 20% enhancement of proliferation of BHK-21 cells at a concentration of 3.40 microM. On the basis of physical and spectral data the structure of compound 1 was established as a tetrapeptide, Gly-Arg-gamma-Glu-Val-NH2.

Amino Acid Sequence↗