Morphological and morphometric study of the androgenetic alopecic scalp using two- and three-dimensional analysis comparing regional differences.

BACKGROUND: Androgenetic (male-type) alopecia (AGA) is caused by genetic and androgenetic effects. The progression of baldness results in smaller hair papillae, thinner hair and a shortened hair cycle. Alopecia occurs mainly in the frontal region and, to a lesser extent, in the occipital region. OBJECTIVES: The morphological differences in the hair follicular units between the alopecic frontal scalp and the vertex and occipital regions were compared using cross-sectional histology and three-dimensional reconstruction. METHODS: Skin specimens were obtained from the frontal, vertex and occipital regions of 24 male human cadavers with fully progressed AGA, and from the frontal region of 32 normal cadaveric scalps. These specimens were fixed, processed using routine histological methods, serially sectioned at a thickness of 10 µm and then stained with Masson's trichrome. The serial sections were reconstructed three-dimensionally using 'Reconstruct' software. RESULTS: The ratios between the numbers of terminal and vellus hairs in the frontal and occipital regions in the AGA scalps were 0·2 : 1 and 3·5 : 1, respectively. Almost all of the hair follicles in the frontal region were vellus hair follicles. The sebaceous gland and arrector pili muscle were larger in the frontal region than in the occipital region. CONCLUSIONS: The morphology of the AGA scalp has been characterized. The terminal-to-vellus hair ratio in the occipital (normal) region was different from that in the frontal (alopecic) region. Moreover, sebaceous glands were larger in the frontal alopecic region than in the occipital region. These larger glands may be associated with other dermatological pathologies, such as seborrhoeic dermatitis.

Purification of an allene oxide synthase and identification of the enzyme as a cytochrome P-450.

Fatty acid hydroperoxides (lipoxygenase products) are metabolized to allene oxides by a type of dehydrase that has been detected in plants, corals, and starfish oocytes. The allene oxides are unstable epoxide precursors of more complex products such as jasmonic acid, the plant growth hormone. Characterization of the dehydrase enzyme of flaxseed revealed that it is a 55-kilodalton hemoprotein. The spectral characteristics of this dehydrase revealed it to be a cytochrome P-450. It operates with the remarkable activity of greater than or equal to 1000 turnovers per second. The results establish a new catalytic activity for a cytochrome P-450 and illustrate the cooperation of different oxygenases in pathways of fatty acid metabolism.

Extra- and intramuscular nerves distributions of the triceps surae muscle as a basis for muscle resection and botulinum toxin injections.

PURPOSE: To compare the distribution of extramuscular nerve branches with their intramuscular ramifications in the triceps surae muscle, thus providing anatomical substantiation for the topography of muscle resection and botulinum toxin injections. METHODS: Dissection and modified Sihler's staining of 18 whole-mount human cadaveric specimens. RESULTS: The distance between the areas with the highest extramuscular branch density and the area of densest intramuscular arborization in gastrocnemius and soleus muscles is approximately 10% of the calf length. This finding should be taken into consideration during nerve blocking and botulinum toxin injections for the treatment of spasticity. Intramuscular nerve arborization patterns make it possible to outline neuromuscular segments in the gastrocnemius and soleus muscles. CONCLUSIONS: Surgical or therapeutic interventions in areas of high extramuscular and intramuscular nerve density can increase the efficacy and safety of botulinum toxin injections and neurotomy. Intramuscular nerve branching patterns should be taken into consideration during triceps surae resection.

Anatomy of the deep branch of the ulnar nerve.

UNLABELLED: The aim of this study was to provide a clear description of the course, precise branching pattern and distribution of the deep branch of the ulnar nerve. A total of 36 hands from 18 preserved cadavers were dissected. The vertical distance from the pisoscaphoid line to the crossing points between the deep branch of the ulnar nerve and each metacarpal was about 4 cm. The deep branch of the ulnar nerve gave off two types of muscular branches: (1) trunks that innervate more than two intrinsic hand muscles; and (2) multiple separate branches innervating only a single muscle. The median numbers of trunks and separate branches were 5 and 6, respectively. LEVELS OF EVIDENCE: N/A.

Properties of Dye-Sensitized Solar Cells Using Carbon Nanowall Counter Electrodes.

This research investigates plasma-treated and metal-coated carbon nanowalls (CNWs) for use as counter electrodes of dye-sensitized solar cells (DSSCs). The CNWs were synthesized on a fluorine-tin-oxide (FTO) glass substrate using the microwave plasma-enhanced chemical vapor deposition (PECVD) system with methane (CH4) gas. The post-plasma treatment was performed on the CNWs with hydrogen (H2) plasma using PECVD, and the CNWs were sputter-coated with metal films using the RF magnetron sputtering system with a four-inch tungsten (W) target. Then the post-plasma-treated and metal-coated CNWs were used as counter electrodes for the fabrication of the DSSCs. Field-emission scanning electron microscopy (FE-SEM) was performed to obtain cross-sectional and planar images of the grown CNWs. The energy conversion efficiencies of the DSSCs manufactured using the post-plasma-treated and metal-layer-coated CNWs as the counter electrodes were measured.

Identification of novel hydroxy fatty acids in the barnacle Balanus balanoides.

The ability of the barnacle Balanus balanoides tissues to produce eicosanoid hatching factors from endogenous polyunsaturated fatty acids has been investigated. GC-MS analysis of an active HPLC fraction from the trihydroxy fatty acid band on TLC revealed the presence of a number of trihydroxy fatty acids and two compounds which were tentatively identified as chlorinated dihydroxy fatty acids. The identified trihydroxy fatty acids are 10,11,12-trihydroxy-5,8,14-eicosatrienoic acid, 10,11,12-trihydroxy-5,8,14,17-eicosatetraenoic acid, 13,14,15-trihydroxy-5,8,11,17-eicosatetraenoic acid and 12,13,14-trihydroxy-4,7,10,16,19-docosapentaenoic acid. The tentatively identified chlorinated dihydroxy fatty acids are 9-chloro- and/or 11-chloro-8,12-dihydroxyeicosatetraenoic acid. The formation of these compounds is evidence of lipoxygenase activities in Balanus balanoides and their identification will facilitate the understanding of the roles eicosanoids play in barnacle physiology, especially with regard to the larval hatching process.

Regulation of estrogen sulfotransferase expression in Leydig cells by cyclic adenosine 3',5'-monophosphate and androgen.

Estrogen sulfotransferase (EST) catalyzes the specific sulfonation and inactivation of estrogens. A common site for EST expression in mammalian species is the testicular Leydig cells. In previous in vivo studies, we have shown that testicular expression of EST is under the regulation of LH. Thus, EST expression in mouse Leydig cells was abolished by hypophysectomy, but could be restored by hCG injection. In this study, we have evaluated the downstream mechanisms by which LH exerts its regulatory effect on EST. Primary mouse Leydig cells were isolated and purified by collagenase digestion and Percoll density gradient centrifugation. They were cultured in serum-free medium at 32 C and treated with various agents for 24 or 48 h, and levels of EST messenger RNA and enzyme activity were determined. Consistent with the in vivo data suggesting an essential role of LH in regulating EST expression, treatment of primary mouse Leydig cells in vitro with 100 microM 8-bromo-dibutyryl cAMP [(Bu)2cAMP] increased EST expression 3- to 5-fold. The effect of (Bu)2cAMP was attenuated by the steroidogenesis inhibitor aminoglutethimide and was mimicked by the potent androgen 5alpha-dihydrotestosterone (5-DHT). The activity of 5-DHT in stimulating EST expression was blocked by the androgen receptor antagonist, hydroxyflutamide. These data suggested the involvement of androgen in (Bu)2cAMP-induced EST expression. Further evidence came from the study with interleukin-1beta, another agent known to suppress Leydig cell steroidogenesis by down-regulating P450c17 gene expression. Treatment of Leydig cells with 0.2 ng/ml interleukin-1beta inhibited (Bu)2cAMP-induced EST expression, which was overcome by the addition of 5-DHT. Finally, in the testis-feminized mouse (Tfm) in which the androgen receptor is nonfunctional due to a frameshift mutation, testicular EST expression is completely absent, whereas messenger RNAs of steroidogenic enzymes such as P450c17 and 3beta-hydroxysteroid dehydrogenase are relatively abundant. We conclude that, by acting as an autocrine or paracrine factor, androgen plays an essential role in the regulation of estrogen sulfotransferase expression in Leydig cell by LH and cAMP.

Estrogen sulfotransferase: discrete and androgen-dependent expression in the male reproductive tract and demonstration of an in vivo function in the mouse epididymis.

Estrogen sulfotransferase (EST) catalyzes the sulfoconjugation and inactivation of the steroid hormone estrogen. It is known previously that EST is expressed abundantly in Leydig cells of the testis. We recently have shown that male mice with targeted EST gene disruption developed age related Leydig cell and seminiferous tubule abnormalities as a consequence of increased local estrogen stimulation. In the same study, we also found that epididymal sperm isolated from the mutant mice had significantly reduced motility, but whether this reflected impaired epididymal function or was secondary to the testicular lesions was not known. The purpose of the current study was to investigate if EST is normally present in the mouse epididymis and/or other parts of the male reproductive tract where, as in testis, it may play a role in regulating local estrogen homeostasis. We describe here that EST is expressed in the epithelium of corpus and cauda but not caput regions of the mouse epididymis. It is also expressed in the luminal epithelium and smooth muscle cells of the vas deferens but was present at very low levels, if at all, in the prostate or seminal vesicle/ coagulating gland. Hypophysectomy, castration, and epididymal ligation experiments, together with the use of an androgen receptor antagonist, established that EST expression in the epididymis and vas deferens is critically dependent on pituitary hormone(s) and androgen but not on other factors in the testicular fluid. Administration of exogenous estradiol to mice with surgically ligated epididymis resulted in a more pronounced reduction in sperm motility in EST mutant mice than in wild-type mice. We conclude that EST is discretely expressed and regulated in the male reproductive tract and plays a physiological role in maintaining the functional integrity of the epididymis by regulating luminal estrogen homeostasis.

Molecular characterization of a testis-specific estrogen sulfotransferase and aberrant liver expression in obese and diabetogenic C57BL/KsJ-db/db mice.

Sulfation represents a major pathway for the inactivation of steroid hormones such as estrogens and is catalyzed by a group of enzymes called sulfotransferases. Aberrant regulation of an estrogen sulfotransferase has been demonstrated previously in the livers of obese and diabetogenic C57BL/KsJ-db/db strain mice. In this paper, we report the molecular cloning and functional characterization of a full-length complementary DNA for estrogen sulfotransferase from mouse testis. The mouse estrogen sulfotransferase complementary DNA encodes 295 amino acids. It shares 88%, 77%, 75%, and 68% identity in amino acid sequence with the rat liver, human liver, guinea pig adrenal, and bovine placental estrogen sulfotransferase, respectively. The mouse enzyme was expressed as a glutathione-S-transferase fusion protein in Escherichia coli. The fusion protein was affinity purified, and milligram quantities of pure enzyme were obtained after cleavage of the fusion protein with thrombin. The expressed enzyme exhibits a high substrate specificity toward estrogens, including estradiol and estrone. Neither dehydroepiandrosterone, pregnenolone, testosterone, nor a simple phenolic compound, 4-nitrophenol appears to be a substrate. Northern hybridization indicates that messenger RNA (1.3 kilobases) for the estrogen sulfotransferase is expressed exclusively in the testes in control C57BL/KsJ mice. However, both the messenger RNA and protein are dramatically induced in the livers of obese and diabetogenic C57BL/KsJ-db/db mice. In contrast to the liver, the constitutive expression of the enzyme in the testis is not affected by the db/db genotype. These results recapitulate the species-specific nature in the tissue distribution of estrogen sulfotransferase and suggest complex regulatory mechanisms in its expression under normal and pathophysiological conditions.

Aberrant cholesterol transport and impaired steroidogenesis in Leydig cells lacking estrogen sulfotransferase.

Estrogen sulfotransferase (EST) is a cytosolic enzyme that catalyzes the sulfoconjugation and inactivation of estrogens. It is expressed abundantly in the mammalian testes in which it may modulate the activity of locally produced estrogen. We demonstrate here that testicular Leydig cells from mice rendered deficient in EST expression by targeted gene deletion acquire a phenotype of increased cholesterol ester accumulation and impaired steroidogenesis with natural aging or in response to estrogen challenge. Abnormal accumulation of cholesterol ester in the mutant Leydig cells correlated with induced expression of the scavenger receptor type B class I, and cultured EST-deficient but not wild-type Leydig cells avidly uptook high-density lipoprotein cholesterol ester ex vivo. EST-deficient Leydig cells in culture produced 50-70% less testosterone than wild-type cells. This deficiency was reversed by androstenedione but not progesterone supplementation, indicating that reduced activities of 17-alpha-hydroxylase-17, 20-lyase were responsible. This conclusion was corroborated by decreased expression levels of 17-alpha-hydroxylase-17, 20-lyase but not of other key steroidogenic enzymes in the mutant cells. These results suggest that EST plays a physiologic role in protecting Leydig cells from estrogen-induced biochemical lesions and provide an example of critical regulation of tissue estrogen sensitivity by a ligand-transformation enzyme rather than through estrogen receptors.

Cellular localization and regulation of expression of testicular estrogen sulfotransferase.

Estrogen sulfotransferase (EST) is a cytosolic enzyme that catalyzes the specific sulfonation of estrogens at the 3-hydroxyl position using 3'-phosphoadenosine-5'-phosphosulfate as an activated sulfate donor. Sulfated estrogens no longer bind to the estrogen receptor and are, therefore, hormonally inactive. Although liver has been considered a primary site for steroid sulfotransferase activities, we previously have cloned the mouse EST complementary DNA and found the enzyme to be expressed abundantly in the testis of normal mice. In this study we show by reverse transcription-PCR that EST is also expressed in the testes of rat and man, suggesting that testicular expression of EST may be a common phenomenon among different species. Using a purified polyclonal antibody raised against the bacterially expressed mouse EST protein, we demonstrate by immunohistochemistry that EST is localized selectively to the androgen-producing Leydig cells within the mouse testis. Additionally, we show that Leydig cell expression of EST is under the control of the pituitary hormone LH and is regulated differentially during development. In contrast to the high level of expression in mature intact animals, EST is not present in Leydig cells of hypophysectomized mice or in Leydig cells of fetal and prepubertal (day 5 or 17) mouse testes. Administration of hCG to hypophysectomized mice restored the testicular expression of EST. Together, these results suggest that testicular expression of EST may play an important role in male reproduction, conceivably by modulating the activity of locally synthesized estrogen in the testis of a sexually mature animal.

Targeted disruption of the mouse estrogen sulfotransferase gene reveals a role of estrogen metabolism in intracrine and paracrine estrogen regulation.

Elicitation of biological responses by estrogen in target tissues requires the presence of ER as well as receptor-active ligand in the local microenvironment. Though much attention has been devoted to the study of the receptor in estrogen target tissues, the concept is emerging that tissue estrogen sensitivity may also be regulated by ligand availability through metabolic transformation in situ. Here, we show that targeted disruption, in the mouse, of an estrogen metabolic enzyme, estrogen sulfotransferase (EST), causes structural and functional lesions in the male reproductive system. EST catalyzes the sulfoconjugation and inactivation of estrogen and is expressed abundantly in testicular Leydig cells. Although knockout males were fertile and phenotypically normal initially, they developed age-dependent Leydig cell hypertrophy/hyperplasia and seminiferous tubule damage. Development of these lesions in the testis could be recapitulated by exogenous E2 administration in younger knockout mice, suggesting that they arose in older knockout mice from chronic estrogen stimulation. Older knockout mice were also found to have reduced testis and epididymis weights but increased seminal vesicle/coagulating gland weight because of tissue swelling. Furthermore, total and forward sperm motility of older knockout mice was reduced by 60% and 80%, respectively, and these mice produced smaller litters compared with age-matched wild-type males. These findings establish a role for EST in the male reproductive system and indicate that intracrine and paracrine estrogen activity can be modulated by a ligand transformation enzyme under a physiological setting. Thus, inhibition of estrogen metabolic enzymes by environmental chemicals, as has been demonstrated recently for the human EST, may constitute a novel mechanism of endocrine disruption in vivo.

Mouse steroid sulfotransferases: substrate specificity and preliminary X-ray crystallographic analysis.

Three mouse cytosolic sulfotransferases were expressed in Escherichia coli cells in order to study their substrate specificities toward natural as well as synthetic steroid hormones. The Km and Vmax values confirmed the high substrate specificity of estrogen and hydroxysteroid sulfotransferases toward estradiol and dehydroepiandrosterone, respectively. In sharp contrast, the synthetic estrogen diethylstilbestrol was metabolized efficiently by both enzymes to its disulfate ester. These sulfotransferases display highly stereospecific sulfotransferase activity for sulfating only the trans-isomer of diethylstilbestrol. Crystals suitable for high-resolution structure determination of estrogen sulfotransferase were grown with polyethylene glycol. The crystals belong to the orthorhombic space group P2(1)2(1)2, and diffracted to 2.5 A.

Correlation between PAP-dependent steroid binding activity and substrate specificity of mouse and human estrogen sulfotransferases.

Estrogen sulfotransferase (EST) is a cytosolic enzyme that catalyzes the sulfoconjugation and inactivation of estrogens using 3'-phosphoadenosine-5'-phosphosulfate (PAPS) as an activated sulfate donor. A finding of undetermined significance in the study of EST has been that the guinea pig EST is able to bind pregnenolone and estradiol with high affinity in the presence of PAP, the reaction by-product of the sulfate donor PAPS. This finding has raised the possibility that EST may have other physiological functions independent of its enzymatic activity as a sulfotransferase. To determine if the PAP-dependent steroid binding activity is a common property shared by other estrogen sulfotransferases, we have expressed the mouse and human EST in bacteria and used the purified protein to address this question. We found that, in the presence of PAP, both recombinant mouse and human EST were able to bind estradiol with high affinity but only the human EST was able to bind pregnenolone. In addition, we show that human but not the mouse EST was also able to bind dehydroepiandrosterone, a property that was not described for the guinea pig EST. Furthermore, we demonstrate that the promiscuity of human EST in steroid binding is mirrored by a correspondingly low substrate specificity in its enzymatic activity as a sulfotransferase. Reversely, the lack of stable binding of pregnenolone and dehydroepiandrosterone by the mouse EST is paralleled by a lack of sulfotransferase activity of this enzyme toward these two steroids. Mutagenesis of mouse EST within a domain critical for PAPS binding abolished both its sulfotransferase and PAP-dependent estrogen binding activity. These data suggest that stable binding of steroids such as pregnenolone or estrogen is not an independent property of estrogen sulfotransferases but rather is related to their catalytic activity.

Biochemistry and reproductive endocrinology of estrogen sulfotransferase.

Estrogen sulfotransferase is a cytosolic enzyme that catalyzes the sulfoconjugation and inactivation of estrogens. Significant progress has been made in the last few years regarding the structure, substrate specificity, tissue expression, and regulation of mammalian estrogen sulfotransferases. The enzyme has high affinity for estrogens and is expressed in a number of estrogen target tissues, including the male and female reproductive systems. Expression of the enzyme in the testis has been particularly well characterized. In the testis, estrogen sulfotransferase is localized selectively to Leydig cells and its expression in these cells is dependent on LH and androgen. It was concluded, from both in vitro and in vivo studies, that estrogen sulfotransferase can function as an effective modulator of local estrogen activity in target tissues. The finding that certain hydroxylated polychlorinated biphenyls are potent inhibitors of the human estrogen sulfotransferase enzyme raises the possibility that environmental chemicals can cause endocrine disruption by enhancing endogenous estrogen activity through inhibition of steroid transformation enzymes such as estrogen sulfotransferase. This provides a new paradigm in explaining the endocrine disrupting potential of environmental chemicals that have low or no binding affinities for steroid hormone receptors.

Early isotonic saline resuscitation from uncontrolled hemorrhage in rats.

BACKGROUND: Attempts to modify traditional fluid resuscitation have been based on animal models that evaluate several variables including anesthesia. This study presents the effects of early saline resuscitation from severe uncontrolled hemorrhage unanesthetized rats. METHODS: Sixty-three female Sprague-Dawley rats were equally divided into three groups: group A, nonresuscitated; and groups B and C, resuscitated ;with isotonic saline (40 and 80 mL/kg, respectively). Hemodynamics, blood loss, survival time, and mortality were recorded for 360 minutes after the hemorrhage, which was initiated by 75% resection of the tail. RESULTS: In group C, 80 mL/kg of saline significantly lowered mortality (24% vs 76% and 71% for groups A and B, respectively) with concomitant increases in mean survival time (241 +/- 103 min vs 146 +/- 108 and 175 +/- 92 min for groups A and B, respectively). There were no statistically significant differences in blood loss, hematocrit, or hemodynamic parameters among the groups. CONCLUSIONS: Early and adequate isotonic saline resuscitation of unanesthetized rats improved outcome despite continuing hemorrhage. The significantly lower mortality rate and increased survival time were not a result of transiently improved arterial pressure and did not correlate with blood loss. No significant bleeding increases were noted in the resuscitated groups.

Membrane complement regulatory proteins: insight from animal studies and relevance to human diseases.

The complement system plays an important role in host defense. However, if not properly regulated, activated complement can also cause significant damage to host tissues. To prevent complement-mediated autologous tissue damage, host cells express a number of membrane-bound complement regulatory proteins. These include decay-accelerating factor (DAF, CD55), membrane cofactor protein (MCP, CD46) and CD59. Recent studies of membrane complement regulatory proteins from various animal species have revealed similarities as well as significant differences from the corresponding human proteins. In this review, we summarize recent advances in this area and contrast the structure, function and tissue distribution of membrane complement regulatory proteins in human and nonprimate mammalian species. We also discuss how the characterization of the animal proteins has provided important clues and might continue to show relevance to the pathogenesis and therapeutics of a number of human diseases.

Topographic anatomy of the zygomatic arch and temporal fossa: a cadaveric study.

The zygomatic arch (ZA) is a long, slender and laterally protruding structure of the face that is vulnerable to fracture by various types of trauma. Knowledge of the topographic anatomy of the ZA and temporal fossa is important for appropriate management of ZA problems. Thirty-seven male and 33 female cadavers were investigated in this study. Skin, subcutaneous tissue, fascia and periosteum were completely removed from around the ZA. Several depths and distances were measured based on three landmarks on the ZA: the anterior, middle and posterior portions of its superior margin. The thickness of the ZA was relatively constant in the three portions. The distance from the internal surface of the ZA to the surface of the temporalis muscle was similar in the anterior and middle portions, at about 8mm, and slightly lesser in the posterior portion. The distance from the external surface of the ZA to the temporal bone was the greatest at the anterior portion, and there was a large difference between the anterior and middle portions. The temporalis muscle was the thickest in the anterior portion and the thinnest in the posterior portion. This study suggests that the maximum distance from the internal surface of the ZA to the surface of the temporalis muscle is 8mm, and this should be considered when performing reduction malarplasty on the ZA.

Topography and location of the depressor anguli oris muscle with a reference to the mental foramen.

The labiomandibular fold (LMF) is the area of the face that extends from the mouth corner to the mandibular border, and its prominence tends to increase with age. The LMF can be formed by the medial or lateral border of the depressor anguli oris (DAO). The aim of this study was to demonstrate the topographical anatomy between the DAO and mental foramen, thereby providing critical information for the safest and most effective site at which to inject botulinum toxin type A (BTX-A). Thirty-four hemifaces from Korean adult cadavers were dissected. The maximum width between the medial borders of the bilateral DAO, parallel to the intercheilion horizontal line, was 59.9 +/- 4.6 (mean +/- SD) mm below the lower lip. The minimum width between the medial borders of the attachment of bilateral DAO was 29.7 +/- 4.8 mm at the mandibular border. The mental foramen was located in the middle third from the cheilion to the mandibular border in 28 cases (90.3%), and it was mostly confined within the DAO muscle coverage in 21 cases (67.7%). The buccal branch of the facial nerve entered through the middle third of the lateral border of DAO and then distributed. Concomitantly, the marginal mandibular branch of the facial nerve entered through the lower third of the lateral border of DAO in 17 cases (60.7%). These results represent additional reference data for identifying the position of the mental foramen on the facial skin, and will be useful for providing criteria for the most effective site for injecting BTX-A when treating the LMF.