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1.
Folia Morphol (Warsz) ; 77(2): 286-292, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29064554

RESUMO

BACKGROUND: Anatomical knowledge of the duodenojejunal flexure is necessary for abdominal surgeries, and also important for physiologic studies about the duodenum. But little is known about the anatomy of this region in mammals. Here, we examined comparative anatomy to understand the anatomical formation of the duodenojejunal flexure in mammals. MATERIALS AND METHODS: The areas around the duonenojejunal flexure were ob-served in mouse, rat, dog, pig, and human, and the anatomical structures around the duodenojejunal junction in the animals were compared with those in human. RESULTS: The superior and inferior duodenal folds, and the superior and inferior duodenal fossae were identified in all examined humans. In pig, the structures were not clearly identified because the duodenum strongly adhered to the retroperitoneum and to the mesocolon. In mouse, rat, and dog, only the plica duodenocolica, which is regarded as the animal counterpart of the superior duo-denal fold in human, was identified, and other folds or fossae were not observed, probably because the duodenum was not fixed to the parietal peritoneum in those animals. Transection of the plica duodenocolica could return the normally rotated intestine back to the state of non-rotation in rat. CONCLUSIONS: This study showed the anatomical similarities and dissimilarities of the duodenojejunal flexure among the mammals. Anatomical knowledge of the area is useful for duodenal and pancreatic surgeries, and for animal studies about the duodenum. (Folia Morphol 2018; 77, 2: 286-292).


Assuntos
Duodeno/anatomia & histologia , Jejuno/anatomia & histologia , Anatomia Comparada , Animais , Cães , Humanos , Ratos , Especificidade da Espécie , Suínos
2.
J Appl Microbiol ; 114(3): 854-60, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23136989

RESUMO

AIMS: To determine the relationship between adhesive ability of probiotics and acidic residues in human colonic mucin, we developed a new screening method using Biacore to evaluate adherence of bacteria before and after sialic acid or sulphate residues were blocked or removed from mucin. METHODS AND RESULTS: Ten strains of lactobacilli and three strains of bifidobacteria isolated from human faeces were evaluated for their adhesive properties to soluble human colonic mucin (sHCM) using the Biacore binding assay. Three strains (Lactobacillus strain ME-522, Lact. gasseri ME-527 and Bifidobacterium bifidum MCC1092) showing significant adherence were selected. Decreased binding activities were observed after removing sialic acid of sHCM using sialidase. However, after removing the sulphate residue using sulphatase, the adhesion of ME-527 decreased; whereas the remaining two strains had increased adhesion. The adhesion of three probiotics significantly decreased after the sulphate residue was blocked by elution with barium chloride. CONCLUSIONS: A new evaluation method using the Biacore assay was developed to observe binding properties to the acidic residues of sHCM. Results indicated that there was a strong relationship between probiotic adhesion and acidic residues of sHCM. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report showing a screening method that quantitatively measures the binding between bacteria and acidic residues in sHCM using the Biacore binding assay; and provides a new method for the selection of probiotics in the future.


Assuntos
Aderência Bacteriana , Bifidobacterium/fisiologia , Lactobacillus/fisiologia , Mucinas/química , Probióticos/análise , Colo/química , Colo/microbiologia , Fezes/microbiologia , Humanos , Ácido N-Acetilneuramínico/química , Sulfatos/química
3.
Radiat Prot Dosimetry ; 146(1-3): 23-6, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21493609

RESUMO

A measurement method for measuring radon in water using the portable radon survey meter (RnSM) was developed. The container with propeller was used to stir the water samples and release radon from the water into the air in a sample box of the RnSM. In this method, the measurement of error would be <20 %, when the radon concentration in the mineral water was >20 Bq l(-1).


Assuntos
Monitoramento de Radiação/instrumentação , Radônio/análise , Poluentes Radioativos da Água/análise
4.
Curr Drug Targets ; 12(5): 621-30, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21039338

RESUMO

ATP-binding cassette (ABC) transporters, P-glycoprotein (P-gp, ABCB1) and ABCG2, are membrane proteins that couple the energy derived from ATP hydrolysis to efflux many chemically diverse compounds across the plasma membrane, thereby playing a critical and important physiological role in protecting cells from xenobiotics. These transporters are also implicated in the development of multidrug resistance (MDR) in cancer cells that have been treated with chemotherapeutics. One approach to blocking the efflux capability of an ABC transporter in a cell or tissue is inhibiting the activity of the transporters with a modulator. Since ABC transporter modulators can be used in combination with chemotherapeutics to increase the effective intracellular concentration of anticancer drugs, the possible impact of modulators of ABC drug transporters is of great clinical interest. Another possible clinical use of modulators that has recently attracted attention is their ability to increase oral bioavailability or increase tissue penetration of drugs transported by the transporters. Several preclinical and clinical studies have been performed to evaluate the feasibility and the safety of this approach. The primary focus of this review is to discuss progress made in recent years in the identification and applicability of compounds that may serve as ABC transporter modulators and the possible role of these compounds in altering the pharmacokinetics and pharmacodynamics of therapeutic drugs used in the clinic.


Assuntos
Transportadores de Cassetes de Ligação de ATP/antagonistas & inibidores , Transportadores de Cassetes de Ligação de ATP/metabolismo , Antineoplásicos/uso terapêutico , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Neoplasias/tratamento farmacológico , Administração Oral , Antineoplásicos/administração & dosagem , Antineoplásicos/metabolismo , Transporte Biológico , Humanos , Terapia de Alvo Molecular , Neoplasias/metabolismo
5.
J Appl Microbiol ; 104(6): 1667-74, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18194256

RESUMO

AIMS: To characterize the adhesion molecule of Lactobacillus plantarum LA 318 that shows high adhesion to human colonic mucin (HCM). METHODS AND RESULTS: The adhesion test used the BIACORE assay where PBS-washed bacterial cells showed a significant decrease in adherence to HCM than distilled water-washed cells. A component in the PBS wash fraction adhered to the HCM and a main protein was detected as a c. 40-kDa band using SDS-PAGE. Using homology comparisons of the N-terminal amino acid sequences compared with sequence databases, this protein was identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The DNA sequence of LA 318 GAPDH was 100% identical to the GAPDH (gapB) of L. plantarum WCFS1. The purified GAPDH adhered to HCM. CONCLUSIONS: We found the adhesin of L. plantarum LA 318 to HCM in its culture PBS wash fraction. The molecule was identified as GAPDH. Because LA 318 possesses the same adhesin as many pathogens, the lactobacilli GAPDH may compete with pathogens infecting the intestine. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report showing GAPDH expressed on the cell surface of lactobacilli adheres to mucin suggesting L. plantarum LA 318 adheres to HCM using GAPDH binding activity to colonize the human intestinal mucosa.


Assuntos
Colo/microbiologia , Gliceraldeído-3-Fosfato Desidrogenases/genética , Mucosa Intestinal/microbiologia , Lactobacillus plantarum/enzimologia , Mucinas/metabolismo , Probióticos , Sequência de Aminoácidos , Aderência Bacteriana , Técnicas Bacteriológicas , Sequência de Bases , Colo/metabolismo , Primers do DNA/genética , Eletroforese em Gel de Poliacrilamida , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Humanos , Mucosa Intestinal/metabolismo , Lactobacillus plantarum/fisiologia , Dados de Sequência Molecular , Mucinas/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Homologia de Sequência de Aminoácidos
6.
Artigo em Inglês | MEDLINE | ID: mdl-12690705

RESUMO

Climbing in the forced swimming test is considered a dopaminergic-specific behavior. A substance of Nicotina tabacum affecting dopamine neuronal activity was investigated using the mouse behavioral system. The substance was found to be a glycoside with the peripheral sugar chain structures Fuc alpha 1-2Gal, Gal beta 1-4GlcNAc and GalNAc alpha 1-3GalNAc and with basic polymannoses. The glycoside dose-dependently increased behavior via D2 neuronal activity, but not D1 activity. This suggests that smoking can affect human brain function not only via the nicotinic cholinergic neuron, but also via the D2 neuron.


Assuntos
Comportamento Animal/efeitos dos fármacos , Glicosídeos/farmacologia , Nicotiana/química , Receptores Dopaminérgicos/fisiologia , Animais , Comportamento Animal/fisiologia , Antagonistas de Dopamina/farmacologia , Glicosídeos/isolamento & purificação , Masculino , Camundongos , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia
7.
Methods Find Exp Clin Pharmacol ; 24(1): 19-21, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-11980383

RESUMO

We investigated the behavioral effects of Chinese herbal medicines in the forced swimming test. One of these medicines, Kami-shoyo-san, induced an antidepressive climbing behavior in mice. An effective substance detected to be an O-linked glycoside with the sugar chain structure GalNAc alpha 1-3GalNAc was separated. The behavioral effect was dose-dependently decreased by the dopamine 2 antagonist sulpiride, but not by the dopamine 1 antagonist SCH-233960. Investigated Chinese herbal medicines, including Kami-shoyo-san, have been used for human depression. These facts suggest that glycoside is one of the antidepressant-like substances of Chinese herbal medicines.


Assuntos
Comportamento Animal/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Glicosídeos/farmacologia , Animais , Relação Dose-Resposta a Droga , Injeções Intraperitoneais , Masculino , Medicina Kampo , Camundongos , Camundongos Endogâmicos , Natação
8.
Tohoku J Exp Med ; 194(4): 223-8, 2001 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11725838

RESUMO

We found a glycoside in sera of schizophrenic patients. This glycoside increased climbing of mouse forced swimming and the climbing was decreased by dopamine D1 receptor antagonist SCH-23390 with a dose dependent manner. This glycoside had much reactivity of GalNAc alpha1-3GalNAc, but not that of polymannoses nor that of Gal beta1-3GlcNAc alpha1-serine or threonine. This strongly suggests that the glycoside is an isolated O-glycoside, but not N-glycoside nor O-glycoside having core 1 or 2. The present findings suggest that schizophrenic patients have the special glycoside affecting the D1 receptor activity in their serum.


Assuntos
Comportamento Animal/efeitos dos fármacos , Comportamento Animal/fisiologia , Glicosídeos/sangue , Glicosídeos/farmacologia , Esquizofrenia/sangue , Adulto , Animais , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Camundongos , Receptores de Dopamina D1/efeitos dos fármacos , Receptores de Dopamina D1/fisiologia , Psicologia do Esquizofrênico , Natação
9.
Nihon Kokyuki Gakkai Zasshi ; 39(5): 363-7, 2001 May.
Artigo em Japonês | MEDLINE | ID: mdl-11510101

RESUMO

The patient was a 69-year-old man with a 3-year history of diabetes mellitus accompanied by cerebral infarction and arteriosclerosis obliterans of the lower extremities. After squatting to clean a car, the patient began to experience dyspnea. A blood gas analysis was performed upon admission, and the patient's PaCO2 and PaO2 levels were found to be low. Examination of a blood sample revealed elevated FDP, FDP-D dimer and platelet factor 4 levels. A perfusion lung scan showed multiple defects in both lungs, but no abnormal findings were detected on a ventilation scan. A pulmonary angiogram showed an interrupted blood flow in branches A9 + 10 of the left pulmonary artery. Accordingly, a pulmonary embolism was diagnosed. Squatting may have caused a partial obstruction of the venous return from the lower extremities by compression of the inguinal region and may have been a predisposing cause of the thrombosis. To our knowledge, such cases of pulmonary embolism triggered by the act of squatting are very rare.


Assuntos
Postura/fisiologia , Embolia Pulmonar/etiologia , Doença Aguda , Idoso , Arteriosclerose Obliterante/complicações , Complicações do Diabetes , Humanos , Masculino , Trombose Venosa/etiologia
10.
Intern Med ; 40(8): 833-5, 2001 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11518138

RESUMO

Eight cases of allergy-like food poisoning resulting from the ingestion of yellowfin tuna, which had been kept in stock for 10 days prior to being cooked, are described. The main symptoms were headaches, facial flushing and palpitation. Samples of the ingested fish were analyzed for histamine content, and a high level of histamine was confirmed (310 mg/100 g of fish). Corticosteroids were given to 3 patients who exhibited dyspnea or persistent symptoms, while the remaining patients improved without medication. In situations where allergy-like clinical features are present after the ingestion of food, the possibility of allergy-like food poisoning should be recognized and included in a differential diagnosis.


Assuntos
Surtos de Doenças , Hipersensibilidade Alimentar/diagnóstico , Hipersensibilidade Alimentar/epidemiologia , Atum/imunologia , Adulto , Animais , Diagnóstico Diferencial , Feminino , Doenças Transmitidas por Alimentos/diagnóstico , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Restaurantes , Atum/microbiologia
11.
Gan To Kagaku Ryoho ; 28(6): 821-4, 2001 Jun.
Artigo em Japonês | MEDLINE | ID: mdl-11432351

RESUMO

Side effects due to administration of anti-cancer drugs often cause the treatment to be abandoned or a decrease in the amount of anti-cancer drugs. Recently, the anti-tumor effects of "low-dose CPT-11", which can be administered at the outpatient clinic, are reported. We performed "low-dose CPT-11 + CDDP" as a neoadjuvant chemotherapy to a patient with advanced gastric cancer. CPT-11 and CDDP combination chemotherapy caused very few side effects, so we could continue the treatment and achieve anti-tumor effects. Consequently, surgery could be performed, but disseminated metastasis was found so that the surgery ended as a non-curative operation. However, it was considered that this method of "low-dose CPT-11 + CDDP" was very effective as the neoadjuvant chemotherapy in a patient with advanced gastric cancer.


Assuntos
Antineoplásicos Fitogênicos/administração & dosagem , Camptotecina/administração & dosagem , Cisplatino/administração & dosagem , Terapia Neoadjuvante , Neoplasias Gástricas/tratamento farmacológico , Idoso , Antineoplásicos Fitogênicos/efeitos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Camptotecina/efeitos adversos , Camptotecina/análogos & derivados , Cisplatino/efeitos adversos , Terapia Combinada , Esquema de Medicação , Humanos , Irinotecano , Masculino , Pessoa de Meia-Idade , Terapia Neoadjuvante/efeitos adversos , Neoplasias Gástricas/cirurgia
12.
Methods Find Exp Clin Pharmacol ; 23(1): 19-21, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11413859

RESUMO

Antidepressants show an antidepressive-like behavioral effect on mouse forced swimming via induction of GalNAc alpha 1-3GalNAc-lipid. In order to detect the mechanisms of the neuron receptor activities on the effect, the glycolipid reactivities in the serum induced by tandospirone, a specific 5-HT1A receptor agonist, and clonidine, a specific alpha 2-adrenoceptor agonist, were investigated. Both of these agonists showed dose-dependent behavioral effects which were decreased by pretreatment with yohimbine, a specific alpha 2-adrenoceptor antagonist. The glycolipid reactivities corresponded with the effects. These present findings strongly suggest that alpha 2-adrenoceptor activity in the brain mediates the induction of the antidepressant glycolipid in the forced swimming test.


Assuntos
Agonistas alfa-Adrenérgicos/farmacologia , Clonidina/farmacologia , Receptores Adrenérgicos alfa/efeitos dos fármacos , Animais , Ansiolíticos/farmacologia , Interações Medicamentosas , Globosídeos/farmacologia , Isoindóis , Masculino , Camundongos , Piperazinas/farmacologia , Pirimidinas/farmacologia , Natação
13.
J Allergy Clin Immunol ; 107(6): 1100-5, 2001 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11398092

RESUMO

BACKGROUND: Food-dependent exercised-induced anaphylaxis (FEIAn) is classified among the physical allergies. Many different food allergens have been reported, but the pathophysiology of FEIAn remains unknown. Furthermore, provocation tests with a suspected food do not always succeed in patients with FEIAn. OBJECTIVE: We sought to clarify and investigate causative foods and mechanisms of FEIAn in a 14-year-old boy. In addition, we tested in vivo and in vitro effects of cromolyn sodium in the same patient. METHODS: We used open challenge tests for the provocation of FEIAn and measured changes in plasma histamine levels and FEV1. In addition, we investigated the mechanism of FEIAn in this case with in vitro histamine release testing. RESULTS: The patient was diagnosed as having FEIAn by provocation testing with a simultaneous intake of wheat and umeboshi, but not when each food was eaten singly, followed by exercise. In addition, his plasma histamine level increased transiently and forced expiratory volume 1, expressed as a percentage change from baseline, decreased significantly. A synergistic effect on in vitro histamine release testing with 2 kinds of the causative foods was shown. Administration of cromolyn sodium proved to be effective on both the in vitro and in vivo tests. CONCLUSION: This is the first report of FEIAn provoked by the test with a simultaneous intake of 2 kinds of food. This case might in part explain negative challenge test results in patients with FEIAn.


Assuntos
Anafilaxia/etiologia , Exercício Físico , Hipersensibilidade Alimentar/complicações , Frutas/efeitos adversos , Triticum/efeitos adversos , Adolescente , Anafilaxia/diagnóstico , Anafilaxia/tratamento farmacológico , Antiasmáticos/uso terapêutico , Cromolina Sódica/uso terapêutico , Hipersensibilidade Alimentar/tratamento farmacológico , Volume Expiratório Forçado/imunologia , Liberação de Histamina , Humanos , Masculino
14.
Curr Opin Neurobiol ; 11(1): 66-73, 2001 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-11179874

RESUMO

Recently, a number of molecules originally thought to have a primary role in cell determination have been shown to affect the cell cycle at specific check points, while other molecules discovered for their roles in the cell cycle progression are known to affect the determination and differentiation of neurons. These discoveries have led to a more detailed investigation of the complex molecular machinery that co-ordinates proliferation and differentiation.


Assuntos
Ciclo Celular/fisiologia , Sistema Nervoso/citologia , Animais , Proteínas de Ciclo Celular/fisiologia , Diferenciação Celular/fisiologia , Humanos
15.
Intern Med ; 40(12): 1245-9, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11813853

RESUMO

We report a 48-year-old man with thrombosis of the portal and superior mesenteric vein and inferior vena cava associated with primary antiphospholipid syndrome (APS). Primary APS was diagnosed by a positive reaction with anticardiolipin antibody (aCL) and the absence of any evidence suggesting the presence of other disease states known to be associated with aCL. A coeliac angiography showed obstruction of the portal and superior mesenteric vein with prominent collaterals and cavernous transformation. Femoral vein angiography showed total obstruction of the external iliac vein and inferior vena cava, and dilation of the pelvic veins, with contrast medium in the lumbar vein. This case is noteworthy as a report of primary APS accompanied by extensive abdominal and pelvic venous thrombosis.


Assuntos
Síndrome Antifosfolipídica/complicações , Síndrome Antifosfolipídica/diagnóstico , Oclusão Vascular Mesentérica/imunologia , Veias Mesentéricas , Veia Porta , Trombose/imunologia , Veia Cava Inferior , Síndrome Antifosfolipídica/diagnóstico por imagem , Humanos , Angiografia por Ressonância Magnética , Masculino , Oclusão Vascular Mesentérica/diagnóstico por imagem , Veias Mesentéricas/diagnóstico por imagem , Pessoa de Meia-Idade , Veia Porta/diagnóstico por imagem , Radiografia , Trombose/diagnóstico por imagem , Veia Cava Inferior/diagnóstico por imagem
16.
Shinrigaku Kenkyu ; 72(5): 369-77, 2001 Dec.
Artigo em Japonês | MEDLINE | ID: mdl-11883324

RESUMO

This study examined how environmental change and others' behavior affected cooperative behavior and solution preference of the person in social dilemma situation. Participants in two experiments played an "environment game," in which gradual pollution in environment and reduction in profit rate were simulated. Information on behavior of other players was manipulated: in "free rider" condition, one person was an extreme free rider, and the others were cooperative; in "loafing" condition, everyone loafed. In both experiments, "Bad Apple Effect" was not observed clearly, and cooperative behavior increased as environmental pollution worsened. In Experiment 2, there was no main effect of others' behavior on solution preference. However, significant correlations were found among solution preference, motivation to control others' behavior, and perceived seriousness of the situation, only when an extreme free rider was among them.


Assuntos
Comportamento Cooperativo , Resolução de Problemas , Problemas Sociais , Adulto , Feminino , Humanos , Masculino , Modelos Psicológicos
17.
Biochim Biophys Acta ; 1529(1-3): 33-48, 2000 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-11111076

RESUMO

Isoprenyl diphosphate synthases catalyze consecutive condensations of isopentenyl diphosphates with allylic primer substrates to form linear backbones for all isoprenoid compounds including cholesterol. These synthases are classified according to the final chain length of their end products and the stereochemistry of the newly formed double bonds. Mutagenesis and X-ray crystallography data have uncovered the basic catalytic and chain length determination mechanisms of E-isoprenyl diphosphate synthases and shed light on their possible evolutionary course. Although much less is known about the Z-isoprenyl diphosphate synthase family, successful cloning and subsequent crystallizations in the near future will no doubt bring more insight as researchers begin to unravel the essential components and precise reaction mechanisms of this cellular machinery.


Assuntos
Alquil e Aril Transferases/metabolismo , Proteínas de Bactérias , Colesterol/biossíntese , Alquil e Aril Transferases/química , Sequência de Aminoácidos , Animais , Bactérias , Catálise , Evolução Molecular , Humanos , Cinética , Modelos Químicos , Estrutura Molecular , Filogenia , Plantas , Estereoisomerismo
18.
Eur J Biochem ; 267(14): 4520-8, 2000 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10880976

RESUMO

A member of the medium-chain prenyl diphosphate synthases, Bacillus stearothermophilus heptaprenyl diphosphate synthase, catalyzes the consecutive condensation of isopentenyl diphosphate with allylic diphosphate to produce (all-E)-C35 prenyl diphosphate as the ultimate product. We previously showed that the product specificity of short-chain prenyl diphosphate synthases is regulated by the structure around the first aspartate-rich motif (FARM). The FARM is also conserved in a subunit of heptaprenyl diphosphate synthase, component II', which suggests that the structure around the FARM of component II' regulates the elongation. To determine whether component II' regulates the product chain length by a mode similar to that of the short-chain prenyl diphosphate synthases, we replaced a bulky amino acid at the eighth position before the FARM of component II', isoleucine 76, by glycine and analyzed the product specificity. The mutated enzyme, I76G, can catalyze condensations of isopentenyl diphosphate beyond the native chain length of C35. Moreover, two mutated enzymes of A79Y and S80F, which have a single replacement to the aromatic residue at the fourth or the fifth position before the FARM, mainly yielded a C20 product. These results strongly suggest that a common mechanism controls the product chain length of both short-chain and medium-chain prenyl diphosphate synthases and that, in wild-type heptaprenyl diphosphate synthase, the prenyl chain can grow on the surface of the small residues at positions 79 and 80, and the elongation is precisely blocked at the length of C35 by isoleucine 76.


Assuntos
Alquil e Aril Transferases/química , Geobacillus stearothermophilus/enzimologia , Alquil e Aril Transferases/genética , Alquil e Aril Transferases/metabolismo , Sequência de Aminoácidos , Cromatografia em Camada Fina , Geobacillus stearothermophilus/genética , Modelos Biológicos , Modelos Moleculares , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Estrutura Terciária de Proteína , Homologia de Sequência de Aminoácidos , Especificidade por Substrato
19.
Clin Nephrol ; 53(2): 90-8, 2000 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-10711410

RESUMO

AIM: To facilitate the understanding of the transporter function of human renal tubular cells, we have developed a simple method using primary cultured proximal tubule (PT) cells isolated from voided urine. METHODS: PT cells grown to confluence on glass coverslips could be identified by parallel arrays of spindle cells and hemicyst formation. Brush-border gamma-glutamyl transpeptidase (gammaGTP) activity was histochemically identified. Apical membrane Na+/H+ exchanger (NHE) activity was measured by monitoring changes in intracellular pH (pHi) after an acid load in a single cell level using the pH-sensitive dye 2'7'-bis-(2-carboxyethyl)-5.6'carboxyfluorescein (BCECF). RESULTS: Amiloride and 5-(N-ethyl-N-isopropyl) amiloride (EIPA) inhibited the NHE activity with half-maximal inhibition values (IC50) of 15.3 and 4.0 microM, respectively. NHE-3 mRNA was detected by the RT-PCR technique in clonally proliferated PT cells. CONCLUSION: These results suggest that cultured PT cells isolated from human urine express amiloride-resistant NHE-3 activity on the apical membranes, which can be compared to functional properties of PT in vivo. Our experimental strategy offers a useful experimental approach to investigating human renal tubular transport function in vitro.


Assuntos
Túbulos Renais Proximais/citologia , Adulto , Sequência de Bases , Transporte Biológico , Técnicas de Cultura de Células/métodos , Células Cultivadas , Pré-Escolar , Primers do DNA , Histocitoquímica , Humanos , Lactente , Recém-Nascido , Túbulos Renais Proximais/metabolismo , Masculino , Microscopia Eletrônica , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Trocadores de Sódio-Hidrogênio/metabolismo , Urina/citologia , gama-Glutamiltransferase/metabolismo
20.
Methods Find Exp Clin Pharmacol ; 22(9): 667-70, 2000 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11294007

RESUMO

GalNAc alpha 1-3GalNAc-lipids induce an antidepressive-like effect in mice in the forced swimming test. In the present study, the relationship between the effects of several antidepressants and serum glycolipid reactivities were investigated. The antidepressants imipramine, maprotiline, paroxetine, trazodone and mianserin induced an effect in forced swimming in mice after 1 or 6 h but not 15 min after treatment. The GalNAc alpha 1-3GalNAc-lipid globopentaosylceramide dose-dependently induced the effect only 15 min after treatment, and the glycolipid reactivities were found in the serum dose-dependently at the period that the mice showed the antidepressive-like climbing behavior. The results strongly suggest that glycolipid production, and not the central neurological activities of the antidepressants, induce the antidepressive effect in mice.


Assuntos
Antidepressivos/farmacologia , Depressão/psicologia , Glicolipídeos/farmacologia , Animais , Antidepressivos/sangue , Relação Dose-Resposta a Droga , Globosídeos/farmacologia , Masculino , Camundongos , Peptídeos/sangue , Natação/psicologia
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