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Background and Objectives@#Treatment choice for fungal ball is endoscopic endonasal removal. However, it is not easy to remove fungal elements from the maxilla using only an endonasal approach. To overcome this difficulty, we introduced a cotton pledget technique and evaluated its efficacy through a cadaveric study and clinical research.Materials and Method: A cadaveric study was performed using 10 half heads of seven cadavers. The ease and safety of the cotton pledget technique were compared to those of a previously reported technique. In clinical research, we enrolled 52 patients who underwent surgery with the cotton pledget technique and 36 patients who underwent surgery using the conventional endoscopic approach. Demographic factors, preoperative Lund-Mackay (LM) score, sinonasal outcome test (SNOT) score, surgical morbidity, and incomplete removal rate were analyzed. @*Results@#The cadaveric study showed that the cotton pledget technique was easier (p=0.011) than the conventional technique. In addition, clinical evaluation showed that the cotton pledget group had significantly lower incomplete removal rate than that of the control group (p=0.010). @*Conclusion@#The cotton pledget technique is an easy and safe method that enables fungal ball removal more effectively than the conventional technique without need for inferior meatal antrostomy (IMA) or the Caldwell-Luc (CL) approach.
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The flexor digitorum superficialis (FDS) muscle is located in the intermediate layer of the muscles in the anterior compartment of the forearm. Variable but individual variations have been reported in the FDS regarding the number of head and the origin, distribution and interconnections of muscle slip and insertion to finger. In this case, we report a concomitant complex variation in FDS which was observed in a cadaver during a routine dissection classes for the undergraduate medical students. It includes the variation which is the separation of the tendon of FDS into the superficial and deep layers, the structural variations in muscle slips and associated tendon variations, the finding of Gantzer' muscle leading to flexor pollicis longus muscle. These complex variations in FDS are very rare case and this report summarizes the related phylogenetic and embryological significance.
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Humanos , Cadáver , Dedos , Antebrazo , Cabeza , Músculos , Estudiantes de Medicina , TendonesRESUMEN
The paper describes a minimally invasive tracheostomy technique that uses an intercartilaginous incision without resection of the tracheal cartilage and discusses its feasibility. A total of 20 adult cadavers (13 males and 7 females) were included in this study. The distance from the arch of the cricoid cartilage to the thyroid isthmus, maximal displacement of the thyroid isthmus, number of tracheal rings underneath the thyroid isthmus, and maximally opened distance resulting from an intercartilaginous incision were measured. The mean distance from the arch of the cricoid cartilage to the thyroid isthmus was 21.4±5.0 mm. The thyroid isthmus mainly overlaid the 3rd and 4th tracheal rings. The mean maximal displacement of the thyroid isthmus was 9.0±2.8 mm. Minimally invasive tracheostomy via an intercartilaginous incision is a feasible technique. A skin incision 2 cm below the cricoid cartilage enables exposure of the thyroid isthmus and anular ligament between the 2nd and 3rd tracheal rings. The intercartilaginous incision allows sufficient space for the tracheostomy tube. An intercartilaginous incision without resection of a tracheal ring can be a good alternative tracheostomy technique, especially for patients who require transient tracheostomy.
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Adulto , Humanos , Masculino , Cadáver , Cartílago , Cartílago Cricoides , Ligamentos , Piel , Glándula Tiroides , Traqueostomía , TraqueotomíaRESUMEN
This study evaluates the suitability of cadavers embalmed by the ethanol-glycerin fixative for the dissection course of medical students and the hands-on dissection workshop of clinicians. Five cadavers were embalmed by two different methods: two formalin-phenol fixation (FPF) and three ethanol-glycerin fixation (EGF) cadavers. The measurement of physical and chemical characteristics including ranges of motion (ROM), bacterial and fungal culture tests, and ultrasonography were performed for each cadaver. The EGF cadavers were evaluated to be significantly more suitable than FPF cadavers for the physical and chemical characteristics including color, texture, elasticity, wetness (softness), skin incision, vessel ligation and suture, decollement, odor, and irritant. In shoulder, elbow, and wrist joints, ROMs of the EGF cadavers were statistically more than those of the FPF except for elbow extension. On bacterial and fungal culture tests at 8 weeks after carrying out of refrigerator, one bacteria were detected in one EGF cadaver; however, some bacteria and fungi could be detected in all FPF cadavers. The ultrasound images of abdominal organ and thigh musculature could be more clearly detected in the EGF cadavers than those of FPF cadavers. These results indicate that the EGF method had a sufficient antibiotic effect and produced cadavers with flexible joints and a high tissue quality suitable for various cadaveric dissection courses.
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Humanos , Bacterias , Cadáver , Educación , Elasticidad , Codo , Embalsamiento , Factor de Crecimiento Epidérmico , Hongos , Articulaciones , Ligadura , Métodos , Odorantes , Hombro , Piel , Estudiantes de Medicina , Suturas , Muslo , Ultrasonografía , Articulación de la MuñecaRESUMEN
To identify genes that participate in the abortion process, normal pregnant uteri were compared to lipopolysaccharide (LPS)-induced abortion uteri. At day 6 of pregnancy, mice were treated with LPS at various time points to induce an abortion. Total RNAs were applied to a cDNA microarray to analyze genes with altered expression. At the early stage (2 hours) of LPS-induced abortion, upregulated genes were mainly composed of immune responsive genes, including Ccl4, Ccl2, Cxcl13, Gbp3, Gbp2, Mx2, H2-Eb1, Irf1 and Ifi203. Genes related to toll-like receptor signaling were also overexpressed. At late stages of abortion (12-24 hours), many genes were suppressed rather than activated, and these were mainly related to the extracellular matrix, cytoskeleton, and anti-apoptosis. Altered expression of several selected genes was confirmed by real time reverse transcription-polymerase chain reaction. The results demonstrated that many known genes were altered in the LPS-treated pregnant uterus, implying that the molecular mechanisms of the genes involved in LPS-induced abortion are complicated. Further analysis of this expression profile will help our understanding of the pathophysiological basis for abortion.
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Animales , Ratones , Embarazo , Citoesqueleto , Matriz Extracelular , Expresión Génica , Perfilación de la Expresión Génica , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN , Receptores Toll-Like , ÚteroRESUMEN
Anatomical variations of the biceps brachii have been described by various authors, but the occurrence of bilateral asymmetric supernumerary heads is rare and has not been reported. We found three accessory heads of the biceps brachii muscle on right arm and an anomalous third head of biceps brachii on left arm. The third, fourth, and fifth heads of right arm originated from the body of humerus at the insertion site of coracobrachialis and inserted into the distal part of biceps brachii short head in order. The third head of left arm originated from humerus at the insertion site of coracobrachialis and combined with the distal part of biceps brachii and continued to the proximal part of common biceps tendon. Understanding the existence of bilateral asymmetric supernumerary heads of biceps brachii may influence preoperative diagnosis and surgery on the upper limbs.
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Brazo , Cabeza , Húmero , Músculos , Tendones , Extremidad SuperiorRESUMEN
A number of acid-base or electrolyte disorders are associated with decreased or increased HCO3- reabsorption in the renal tubules. The present study was to examine the alterations of expression and distribution of Carbonic anhydrase II in the kidneys of normal and potassium-depleted rats using Western blot analysis and immuno-histochemistry. Western blot analysis demonstrated that CA II protein, ~30 kDa at molecular mass, was abundantly expressed in normal group. All potassium-depleted groups showed slightly increased CA II protein compared to normal group. In control group, immunoreactivity of CA II protein was detected in the entire collecting duct. Signal intensity was prominent in the intercalated cells and weak in the principal cells of the cortical collecting ducts. In potassium-depleted groups, the pattern of cellular labeling of CA II protein was identical to that of normal group, but the signal intensity was decreased in cortical collecting duct, markedly increased in the inner stripe of outer medullary and inner medullary collecting ducts, and unchanged in the outer stripe of outer medullary collecting duct. These results suggest that chronic hypokalemia impact the expression pattern of CA II protein depending the portion of the collecting duct.
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Animales , Ratas , Western Blotting , Carbono , Anhidrasa Carbónica II , Anhidrasas Carbónicas , Hipopotasemia , Inmunohistoquímica , RiñónRESUMEN
Homeobox genes seem to play critical roles in regulating morphogenesis, patterning, organogenesis, and differentiation. They have the conserved sequence that codes the DNA-binding domain called homeodomain. The expression and cellular localization of rPsx mRNA in rat placenta during placental development were examined by in situ hybridization histochemistry at different embryonic stages (Embryonic days 7.5~16.5). rPsx mRNA was first detected in chorionic ectoderm of placenta at E 10.5. This transcript was localized in labyrinth trophoblast and trophoblast giant cells at E 11.5. Hybridization signals were observed in labyrinth trophoblast, spongiotrophoblast, and trophoblast giant cells at E 12.5, E 13.5, and E 14.5. At E 15.5, hybridization signal was detected in labyrinth trophoblast and spongiotrophoblast but not in trophoblast giant cells. Hybridization signal was only detected in labyrinth trophoblast at E 16.5. rPsx mRNA was not detected in decidua and any tissues of the embryo from E 7.5 to E 9.5 of gestations. From these results, a new rPsx homeobox gene is first expressed at E 10.5 and detected in chorionic ectoderm, labyrinth trophblast, spongiotrophoblast and trophoblast giant cells of the placenta. This gene may play a critical role in differentiation and development of trophoblast cells.
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Animales , Femenino , Ratas , Quimera , Corion , Secuencia Conservada , Decidua , Oído Interno , Ectodermo , Estructuras Embrionarias , Expresión Génica , Genes Homeobox , Células Gigantes , Hibridación in Situ , Morfogénesis , Organogénesis , Placenta , Placentación , ARN Mensajero , TrofoblastosRESUMEN
During the prostate cancer (PCa) development and its progression into hormone independency, androgen receptor (AR) signals play a central role by triggering the regulation of target genes, including prostate-specific antigen. However, the regulation of these AR-mediated target genes is not fully understood. We have previously demonstrated a unique role of HOXB13 homeodomain protein as an AR repressor. Expression of HOXB13 was highly restricted to the prostate and its suppression dramatically increased hormone-activated AR transactivation, suggesting that prostate-specific HOXB13 was a highly potent transcriptional regulator. In this report, we demonstrated the action mechanism of HOXB13 as an AR repressor. HOXB13 suppressed androgen-stimulated AR activity by interacting with AR. HOXB13 did neither bind to AR responsive elements nor disturb nuclear translocation of AR in response to androgen. In PCa specimen, we also observed mutual expression pattern of HOXB13 and AR. These results suggest that HOXB13 not only serve as a DNA-bound transcription factor but play an important role as an AR-interacting repressor to modulate hormone-activated androgen receptor signals. Further extensive studies will uncover a novel mechanism for regulating AR-signaling pathway to lead to expose new role of HOXB13 as a non-DNA-binding transcriptional repressor.
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Anafilaxis Cutánea Pasiva , Próstata , Antígeno Prostático Específico , Neoplasias de la Próstata , Receptores Androgénicos , Proteína Estafilocócica A , Factores de Transcripción , Activación TranscripcionalRESUMEN
This study presents distribution of carbonic anhydrase (CA) isozymes IV and IX, membrane associated forms, and CA I and II, cytoplasmic forms, in rat parotid and submandibular glands using Western blot analysis and immunohistochemical staining. Western blot analysis demonstrated that CAs I, II and IX were found to be abundantly expressed, but CA IV was weakly expressed in parotid gland. Submandibular gland expressed abundant CAs I and II, weak CA IX, and undetectable level of CA IV. In hematoxylin-eosin staining, parotid gland was entirely composed of serous acini and their ducts while submandibular gland was mixed population of serous and mucous lobules. Most of lobules (submandibular gland proper type) contained mostly serous acini and their ducts with granular convoluted duct. Some lobules (sublingual gland type) contained mostly mucous acini with serous demilune and their ducts without granular convoluted duct. In parotid gland, CAs IV and IX were immunolocalized in duct cells and not in serous acinar cells. Immunoreactivity for CAs I and II was also detectable in duct cells. Serous acinar cells were positive for CA II, and negative for CA I. In submandibular gland, CAs IV and IX were immunolocalized in duct cells but not in acinar cells of both types of lobules. Immunoreactivity for CAs I and II was also detectable in duct cells of both types of lobules. Cells of serous acini and serous demilune were positive for CA II, and negative for CA I. Mucous cells were negative for both CAs I and II. These results demonstrate the distribution of CA isoenzymes in parotid and submandibular glands of the rat, and suggest CAs IV and IX as well as CAs I and II are related to electrolytes metabolism of saliva in duct cells.
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Animales , Ratas , Células Acinares , Western Blotting , Carbono , Anhidrasas Carbónicas , Citoplasma , Electrólitos , Inmunohistoquímica , Isoenzimas , Membranas , Glándula Parótida , Saliva , Glándulas Salivales , Glándula SubmandibularRESUMEN
There are several carbonic anhydrase (CA) isozymes, which differ in their kinetic properties, tissue distribution, and subcellular localization. In this study, the distribution of CA isozymes I, II, IV, and IX was investigated in the rat exorbital lacrimal gland using Western blotting analysis and immunohistochemical staining. In the Western blotting analysis of the rat lacrimal gland, CA II and CA IX were expressed abundantly and CA IV was expressed weakly. Hematoxylin-eosin staining of the exorbital lacrimal gland showed a multilobular tubuloacinar gland composed of acinar and ductal cells. Immunohistochemical reaction revealed no CAI staining in acinar cells and positive staining in intercalated and small duct cells. CA II reactivity was detected in the supranuclear cytoplasm of acinar cells and appeared to vary between acini. The intercalated and collecting duct cells showed weak or no immunoreactivity for CA II. CA IV was detected in the intercalated and collecting duct cells but not at the acinar cells. CA IX was detected in the intercalated and collecting duct cells, and in only a few acinar cells. These results demonstrate the differential distribution of CA isoenzymes in the exorbital lacrimal gland of the rat and suggest that CA II is related mainly to the electrolyte metabolism of tears in the acinar cells and that CAs I, IV, and IX are related to the electrolyte metabolism of tears in the duct cells.
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Animales , Ratas , Células Acinares , Western Blotting , Carbono , Anhidrasas Carbónicas , Citoplasma , Inmunohistoquímica , Isoenzimas , Aparato Lagrimal , Distribución TisularRESUMEN
Carbonic anhydrase catalizes the reversible hydration of carbonic dioxide and participate in various biological processes. There are several isozymes and differ in their kinetic properties, tissue distribution and subcellular localization. The expression of carbonic anhydrase isozymes in digestive tract vary according to animal species and region of the tract. The distribution of carbonic anhydrase (CA) isozymes I, II, IV and IX was investigated in various portions of the rat small intestine using Western blotting analysis and immunohistochemical staining. Western blotting analysis of rat small intestine revealed that CAI was found to be abundantly expressed throughout the small intestine. Expression of CAII in duodenum was much higher than that in jejunum and ileum. Expression of CAIV and IX was found to be weak throughout the small intestine. Immunohistochemical reaction revealed no staining of CAI in all parts of small intestine except blood vessels. CAII was detected at the supranuclear cytoplasm of surface epithelium, but not in intestinal gland. Staining intensity was most strong in the proximal duodenum. CAIV was detected at the apical surface of epithelial cells of villi, and showed most strong staining intensity in the terminal ileum. CAIX was detected at the surfcae epithelium, cells of intestinal gland and Brunner's gland, and the positive reaction was confined to the supranuclear cytoplasm. CAIX differed from CAII in tissue distribution, but subcellular localization of CAIX and II were the same. These results indicate that the surface epithelium of small intestine express CAII, IV and IX, intestinal gland and Brunner's gland express CAIX, and suggest that CAIX may somewhat contribute the control of acid-base balance in the small intestine.
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Animales , Ratas , Equilibrio Ácido-Base , Fenómenos Biológicos , Vasos Sanguíneos , Western Blotting , Carbono , Anhidrasas Carbónicas , Citoplasma , Duodeno , Células Epiteliales , Epitelio , Tracto Gastrointestinal , Íleon , Inmunohistoquímica , Mucosa Intestinal , Intestino Delgado , Isoenzimas , Yeyuno , Distribución TisularRESUMEN
The distribution of carbonic anhydrase (CA) isoenzymes I, II, IV, and IX was investigated in pancreatic islet of the rat using Western blotting analysis and immunohistochemistry. Western blotting analysis demonstrated strong CAI and II expression, but weak CAIV and no CAIX expression. Immunohistochemical reaction of pancreatic islet revealed no staining for CAI and II. CAIV was detected in the peripheral cells of the islet. CAIX was detected in the peripheral cells and occasional in the centrally located cells. Signals for CAIV were observed at the plasma membrane and/or in the cytoplasm of islet cells. Location of CAIV in the A cells was confirmed by subjecting serial sections of pancreas to staining for CAIV and glucagon, which showed colocalization in the A cells. Immunohistochemical staining of pancreatic acinus revealed abundant staining for CAI in interacinar blood vessels and CAII in ductal and acinar cells. These results demonstrate the differential distribution of CA isoenzymes in pancreatic islet, and suggest that A cells of pancreatic islet might contain both CAIV and IX.
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Animales , Ratas , Células Acinares , Vasos Sanguíneos , Western Blotting , Carbono , Anhidrasas Carbónicas , Membrana Celular , Citoplasma , Glucagón , Inmunohistoquímica , Islotes Pancreáticos , Isoenzimas , PáncreasRESUMEN
The marked hemodynamic and hormonal changes of normal pregnancy are associated with striking alterations in renal physiology involving structure, dynamics, tubular function, and volume homeostasis. A number of acid-base or electrolyte disorders are associated with decreased or increased HCO3-reabsorption in the renal tubules. The present study was to examine the alterations of expression and distribution of Na+/HCO3-cotransporter (NBC), Na+/H+ exchanger-3 (NHE-3), and carbonic anhydrase I and II (CA I, II) proteins in the kidneys of non-pregnant (NP) and pregnant rats using Western blot analysis and immunohistochemistry. Sprague-Dawley female rats were studied on days 10 (P 10), 12 (P 12), 14 (P 14), 17 (P 17), and 19 (P 19) of pregnancy. Western blot analysis demonstrated that the expression of NBC, ~110 kDa at molecular mass, was increased in pregnant rats, particularly P 12, compared with NP rat. The expression of NHE-3, ~83 kDa at molecular mass, was increased in pregnant rats, particularly P 12 and P 14. The expression of CA I, ~30 kDa at molecular mass, was decreased in pregnant rats, particularly P 14, but, CA II protein, ~30 kDa at molecular mass, was similar NP rat. In immunohistochemistry, strong immunoreactivity of NBC of NP rat was exclusively detected in the basolateral membranes of S1 and S2 segment of proximal tubules whereas not in S3 segment. In pregnant rats, the pattern of cellular labeling of NBC was identical to that of NP rat, but signal intensity was increased, particularly P 12. In NHE-3, strong immunoreactivity was detected in apical membranes and brush borders of S3 segments and moderate in S1 and S2 segments. In pregnant rats, the pattern of cellular labeling was identical to that of NP rat, but the signal intensity was increased, particularly P 12 and P 14. Expression of CA I and II proteins was detected in entire collecting duct. Signal intensity was prominent in type A intercalated cells and moderate in type B intercalated cells. In pregnant rats, the pattern of cellular labeling of CA I and II proteins was identical to that of non-pregnant rat, but the signal intensity of CA I was decreased in cortical collecting duct, particularly P 14 and CA II was identical to that of NP rat. These results suggest that the regulation of NBC and NHE-3 expressions in the proximal tubules and CA I expression in cortical collecting duct may maintain HCO3-concentration during the pregnancy.
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Animales , Femenino , Humanos , Embarazo , Ratas , Bicarbonatos , Western Blotting , Anhidrasa Carbónica I , Hemodinámica , Homeostasis , Inmunohistoquímica , Riñón , Membranas , Microvellosidades , Fisiología , Ratas Sprague-Dawley , Control Social Formal , Huelga de EmpleadosRESUMEN
The potassium depletion has remarkable and opposite effect on kidney and body growth and has affected the expression of the several ion transporters. Previously, Ahn et al. have reported that HK alpha 1 and 2 subunit gene were upregulated in the hypokalemic rat kidney. To clone the unreported genes expressed in potassium deficiency, differential display PCR-based cloning strategy was used in normal and potassium-depleted rat kidney and a novel gene was isolated. Sequence analysis with blast search program identified a cDNA clone encoding an isoform of kidney sodium bicarbonate cotransporter-1. The tissue and cellular expression pattern of this gene were investigated with Northern analyses and in situ hybridization histochemistry (ISH) in normal and hypokalemic rats. This novel transcript was highly expressed in kidney and brain and at lower levels in distal colon, urinary bladder, and heart but not in salivary gland, stomach, liver, and lung in normal rat. In potassium-depleted rat, this transcript was upregulated in kidney, brain, and distal colon. By ISH, cellular distribution of this gene was highly expressed in S3 segment of proximal tubule, distal convoluted tubule, and cortical collecting duct of kidney and lower third of intestinal glands of distal colon but at lower levels in cortical and medullary thick ascending limb and medullary collecting duct of kidney and middle third of intestinal glands of distal colon. From these results, this candidate gene may play an important role in HCO3-transport by these organs during potassium depletion.
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Animales , Ratas , Encéfalo , Células Clonales , Clonación de Organismos , Colon , ADN Complementario , Extremidades , Corazón , Hipopotasemia , Hibridación in Situ , Mucosa Intestinal , Transporte Iónico , Riñón , Hígado , Pulmón , Potasio , Deficiencia de Potasio , Glándulas Salivales , Análisis de Secuencia , Bicarbonato de Sodio , Simportadores de Sodio-Bicarbonato , Estómago , Vejiga UrinariaRESUMEN
The most commonly reported sexual problems in diabetic women are sexual arousal disorder and a lack of vaginal lubrication. The aims of this study were to investigate the vaginal structural changes and expressions of TGF-beta1, Ec-NOS and estrogen receptor alphaby histochemistry, immunohistochemistry and Western blot analysis in diabetic and insulin-treated diabetic rats. The mean blood glucose levels were significantly increased in the diabetic rats (453+/-88.4 mg/dL)compared to the control group (79+/-6 mg/dL)and insulin-treated diabetic rats (56.7+/-0.6 mg/dL).The vaginal wall in control rat showed 6~11 layered stratified squamous epithelial lining and submucosal smooth muscle, connective tissue and vasculatures. In diabetic rat, the vaginal epithelium was reduced to 2~6 layers and the submucosal vasculatures were decreased n size and number.Collagen fibers were increased and irregularly distorted arrangement. Insulin-treated diabetic rat showed similar morphologic features as control rat.In diabetic rat, TGF-beta1 expression was upregulated by 1.65 times and Ec-NOS expression was 40% downregulated compared to control and insulin-treated diabetic rats in Western blot analysis. In control and insulin-treated diabetic rats, TGF-beta1 immunoreactivity was detected in fibroblasts and the collagen fibers, Ec-NOS immunoreactivity in the endothelial cells of blood vessels, and estrogen receptor alphaimmunoreactivity in the basal and intermediate cell layers of stratified squamous epithelium, smooth muscle fibers, and nerve fibers. In diabetic rat, expression of TGF-beta1, Ec-NOS, and estrogen receptor alphawas exhibited comparable cellular patterns of labeling, but signal intensity was increased in TGF-beta1 and decreased in Ec-NOS and estrogen receptor alpha. These results suggest that vaginal tissue fibrosis in diabetes mellitus may be caused by altered expression of TGF-beta1, NOS and estrogen. It also mplies that sexual arousal disorder and lack of vaginal lubrication in the diabetic women could be protected or delayed by controlling blood glucose levels.
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Animales , Femenino , Humanos , Ratas , Glucemia , Vasos Sanguíneos , Western Blotting , Colágeno , Tejido Conectivo , Diabetes Mellitus , Células Endoteliales , Epitelio , Receptor alfa de Estrógeno , Estrógenos , Fibroblastos , Fibrosis , Inmunohistoquímica , Insulina , Lubrificación , Músculo Liso , Fibras Nerviosas , Óxido Nítrico Sintasa , Disfunciones Sexuales Psicológicas , Factor de Crecimiento Transformador beta1 , VaginaRESUMEN
Excess accumulation of glucocorticoid increases acid secretion and HCO3- reabsorption in the kidney. Reabsorption of HCO3-, which almost occurs at the proximal tubule, is mediated Na+ / H+ exchanger-3 (NHE-3) and H+ -ATPase on the apical membrane and the Na + /HCO3- cotransporter-1 (NBC-1)on the basolateral membrane. Impact of glucocorticoid was investigated by immunohistochemistry and electron microscopy to correlate the changes with the effect of in vivo dexamethasone treatment for the rat kidney proximal tubule. In a control group, immunoreactivity of NHE-3 was detected in the apical membrane and the brush borders of S1, S2 and 3 segments of the proximal tubule. Immunoreactivity of NBC-1 was detected in the basolateral membrane of S1 and S2 segments of the proximal tubule. Immunoreactivity of NHE-3 and NBC-1 protein was more pronounced in dexamethasone treated groups than the control group. Dexamethasone 1 mg/kg caused most intense immunoreactivity for NHE-3 and NBC-1 protein, however, 0.01 mg/kg and 0.1 mg/kg produced less intense immunoreactivity with no appreciable differences between these lower doses of dexamethasone groups. By electron microscopy, the tubular cells of S1 segment of the control group revealed numerous mitochondria, endocytic apparatuses, lysosomes and many basal cytoplasmic processes. In dexamethasone treated groups, the cells of S1 and S2 segments of the proximal tubule had more mitochodria and more basolateral invaginations and had an increased number of more elongated microvilli, compared with the control group. The cells of the S3 segment of the control group showed scant lateral interdigitations and had a few smaller mitochondria. The cells of the S3 segment of dexamethasone treated groups had many mitochodria and an increased number of microvilli in the brush border, but revealed no difference of basolateral invaginations among the different groups of dexamethasone. These results indicate that prolonged administration of excess glucocorticoid increases NHE-3 and NBC-1 protein, and the up-regulation of these proteins could result in increased HCO3 - reabsorption in the rat renal proximal tubules. It also suggests that these adaptive responses closely correlate to morphological alterations of proximal tubular epithelial cells.
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Animales , Ratas , Citoplasma , Dexametasona , Células Epiteliales , Inmunohistoquímica , Riñón , Lisosomas , Membranas , Microscopía Electrónica , Microvellosidades , Mitocondrias , Regulación hacia ArribaRESUMEN
A number of acid-base or electrolyte disorders are associated with decreased or increased HCO3- reabsorption in the renal tubules. There has been a general agreement that potassium depletion induces metabolic alkalosis and affects the expression of the several ion transporters in rats. The present study was to examine the alterations of expression and distribution of COX-1, 2 mRNAs and proteins in the kidneys of normal and K-depleted rats using RT-PCR, Western blot analysis, and immunohistochemistry. Predicted size of COX-1 mRNA was 306 bp. It's expression was increased in K-depleted rats, particularly LK 2W, but decreased in LK 3D. Predicted size of COX-2 mRNA was 356 bp and it's expression was increased in K-depleted rats, particularly LK 2W. Western blot analysis demonstrated that COX-1 protein, ~70 kDa at molecular mass, was increased in potassium-depleted rats, particularly LK 2W and decreased in LK 3D, compared with normal rat. COX-2 protein, ~72 kDa at molecular mass, was only increased in LK 3D and others were comparable with normal rat. In immunohistochemistry, COX-1 was detected in entire collecting duct, intraglomerular mesangial cells, arterial endothelial cells, medullary interstitial cells, papillary epithelial cells, and pelvic epithelium. Signal intensity of the collecting duct was more increased toward the papillary tip. In K-depleted rat, the pattern of cellular labeling of COX-1 protein was identical to that of normal rat. However, the signal intensity of LK 3D was only decreased in cortical and outer medullary collecting duct and that of LK 2W was increased particularly in the inner stripe of outer medullary collecting duct and proximal 1/3 inner medullary collecting duct. Immunoreactivity of COX-2 of normal rat was detected in the cortical thick ascending limb and macula densa. In K-depleted rat, the pattern of cellular labeling of COX-2 protein was identical to that of normal rat, but the signal intensity was only increased in LK 3D rat. These results suggest that chronic hypokalemia enhances the expression of COX-1, 2 mRNAs and proteins and the regulation of K reabsorption depends on COX-1 rather than COX-2 by the portions of expression.
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Animales , Ratas , Alcalosis , Western Blotting , Ciclooxigenasa 1 , Células Endoteliales , Células Epiteliales , Epitelio , Extremidades , Hipopotasemia , Inmunohistoquímica , Transporte Iónico , Riñón , Células Mesangiales , Potasio , Prostaglandina-Endoperóxido Sintasas , ARN MensajeroRESUMEN
The distribution of carbonic anhydrase (CA) isozymes I, II, IV, and IX was investigated in the human duodenum and colon using Western blotting analysis and immunohistochemical staining. A Western blotting analysis revealed an abundant expression of CAI and IX in the duodenum and colon. The expression of CAII and IV was detected in mucosa of duodenum and colon. The degree of expression, however, showed regional difference. The expression of CAII was strong in the duodenum and colon, and that of CAIV was weak in the duodenum and colon. Immunohistochemical staining of duodenum revealed no staining for CAI. CAII was detected at the columnar cells of surface epithelium and secretory cells and ductal cells of Brunner's gland. CAIV was detected at the ductal cells of Brunner's gland. CAIX was detected at the cells of intestinal gland and rare cells of the Brunner's gland. Immunohistochemical staining of colon showed a positive reaction of CAI and II at the columnar and goblet cells of surface epithelium, and CAIV and IX at the columnar cells of surface epithelium. These results demonstrate the differential distribution of CA isozymes in duodenum and colon, and suggest that Brunner's gland may contribute the control of acid-base balance in the duodenal lumen by secreting bicarbonate ion catalyzed by CAII and IV.
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Humanos , Equilibrio Ácido-Base , Bicarbonatos , Western Blotting , Carbono , Anhidrasas Carbónicas , Colon , Duodeno , Epitelio , Células Caliciformes , Inmunohistoquímica , Mucosa Intestinal , Intestinos , Isoenzimas , Membrana MucosaRESUMEN
Cyclooxygenase (COX)-1 and -2 expressions in the incisional wound healing of mouse skin were determined by immunohistochemistry and Western blot analysis. By Western blotting, compared to normal skin, COX-2 activity was increased at days 1, 4, 8, and 12 and was maximal at 4 day after incisional wound of mouse skin whereas COX-1 was barely detectable. In normal skin, COX-1 immunostaining was observed among the basal cells of epidermis whereas COX-2 immunostaining was detected in the more differentiated, suprabasal keratinocytes. At 1~4 days after wound, COX-2 staining was particularly prominent in the inflammatory cells, and at day 8, many macrophage-like cells were stained positively. COX-2 immunoreactive fibroblast, macrophage-like cells, and newly formed vascular endothelial cells were increased in number at 12 days after incision. These data suggest that COX-2 is constitutively expressed, just as is COX-1, in epidermis and is associated with keratinocyte differentiation. In addition, these findings support the well-established role for COX-2, the prostaglandins that they generate, as mediators of inflammatory response.