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1.
Vet Res Commun ; 45(4): 293-304, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34219191

RESUMO

Wounds are damaging to quality life of confined animals, causing dysfunction in spinal, members injuries, and reduction in productive performance. This research investigated the clove antimicrobial and antioxidant activity on the healing of decubitus wounds (pododermatitis) of rabbits (Oryctolagus cuniculus). Adult animals were treated for 21 days every three days with a fluid gel spray in the wound region: control fluid gel without addition of clove (FGC0), fluid gel with addition of 1% clove powder (FGC1), and fluid gel with 2% clove powder (FGC2). Microbiological analysis for Escherichia coli and Pseudomonas spp. were performed during 21 days of experimental period. After this period, samples from treated skin were evaluated for histological analysis and evaluation of the healing process by spectroscopy (FTIR-ATR). Rabbits treated with FGC2 showed advanced healing and decreased tissue inflammation similar to healthy rabbits, while FGC0 rabbits showed a decrease in bacterial contamination without signs of healing. Both FGC1 and FGC2 rabbits demonstrated antimicrobial and antioxidant action against both bacteria tested, favoring the wound healing process. Considering the results, the use of fluid gel with 2% of clove powder (Syzigium aromaticum) based on the best antimicrobial, antioxidant and anti-inflammatory activities on healing of decubitus wounds (pododermatitis) of rabbits in commercial farming system.


Assuntos
Antibacterianos/farmacologia , Antioxidantes/farmacologia , Doenças do Pé/veterinária , Géis/uso terapêutico , Úlcera por Pressão/veterinária , Coelhos , Syzygium/química , Animais , Dermatite/etiologia , Dermatite/prevenção & controle , Dermatite/veterinária , Feminino , Doenças do Pé/etiologia , Doenças do Pé/prevenção & controle , Masculino , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Úlcera por Pressão/etiologia , Úlcera por Pressão/prevenção & controle , Coelhos/lesões , Cicatrização
2.
Lymphology ; 49(2): 93-106, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29906367

RESUMO

Patients with breast cancer-related lymphedema (BCRL) need a life-long self-care program that they can adhere to enable them to manage their lymphedema. The objective of this study was to assess the effectiveness of a holistic BCRL self-care program that patients could easily adhere to and comply with. A prospective, longitudinal, comparative study between affected arms and unaffected arms in unilateral breast cancer patients was implemented over a six-month period. Both the lymphedematous and unaffected arms of 23 patients with unilateral BCRL were followed and measured. The daily 10-minute holistic BCRL self-care program consisted of modified Japanese rajio taiso (Japanese radio calisthenics), a gentle arm exercise combined with deep breathing, skin moisturizing care using a traditional lymphatic drainage technique, and basic self-care education. Arm and edema volume, relative volume change, resistance of the skin to compression (fibrosis), lymphedema-related symptoms, skin condition, and self-care were assessed. At the end of six-months the volume of all limb segments and resistance of the tissues to compression at all measurement points of the affected arm were significantly reduced. On the unaffected side, only the volume of the forearm and the whole arm was significantly reduced and fibrosis significantly reduced only in the forearm. There was no significant difference in edema volume and relative volume change. Lymphedema-related symptoms significantly improved. Perceived adherence, effectiveness, burden, score and average time for self-care significantly increased. Our results demonstrate that this 10-minute self-care program may improve BCRL and its self-care.


Assuntos
Linfedema Relacionado a Câncer de Mama/terapia , Neoplasias da Mama/cirurgia , Exercícios Respiratórios/métodos , Terapia por Exercício/métodos , Excisão de Linfonodo , Drenagem Linfática Manual/métodos , Mastectomia , Autocuidado/métodos , Higiene da Pele/métodos , Atividades Cotidianas , Adulto , Idoso , Aromaterapia , Axila , Quimioterapia Adjuvante , Citrus paradisi , Feminino , Humanos , Japão , Estudos Longitudinais , Mastectomia Segmentar , Pessoa de Meia-Idade , Óleos Voláteis/uso terapêutico , Cooperação do Paciente , Educação de Pacientes como Assunto , Estudos Prospectivos , Radioterapia Adjuvante , Fatores de Tempo , Resultado do Tratamento
3.
Phytochemistry ; 56(7): 649-55, 2001 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-11314949

RESUMO

S-Adenosyl-L-methionine (SAM): coclaurine N-methyltransferase (CNMT), which catalyzes the transfer of a methyl group from S-adenosyl-L-methionine to the amino group of the tetrahydrobenzylisoquinoline alkaloid coclaurine. was purified 340-fold from Coptis japonica cells in 1% yield to give an almost homogeneous protein. The purified enzyme, which occurred as a homotetramer with a native Mr of 160 kDa (gel-filtration chromatography) and a subunit Mr of 45 kDa (SDS-polyacrylamide gel electrophoresis), had an optimum pH of 7.0 and a pI of 4.2. Whereas (R)-coclaurine was the best substrate for enzyme activity, Coptis CNMT had broad substrate specificity and no stereospecificity CNMT methylated norlaudanosoline, 6,7-dimethoxyl-1,2,3,4-tetrahydroisoquinoline and 1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline. The enzyme did not require any metal ion. p-Chloromercuribenzoate and iodoacetamide did not inhibit CNMT activity, but the addition of Co2+, Cu2+ or Mn2+ at 5 mM severely inhibited such activity by 75, 47 and 57%, respectively. The substrate-saturation kinetics of CNMT for norreticuline and SAM were of the typical Michaelis-Menten-type with respective Km values of 0.38 and 0.65 mM.


Assuntos
Metiltransferases/isolamento & purificação , Metiltransferases/metabolismo , Plantas Medicinais/enzimologia , Alcaloides/metabolismo , Células Cultivadas , Cromatografia em Gel , Cromatografia por Troca Iônica , Isoquinolinas/metabolismo , Cinética , Metiltransferases/química , Peso Molecular , Plantas Medicinais/citologia , Estereoisomerismo , Especificidade por Substrato
4.
Int J Urol ; 7(2): 35-40, 2000 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-10710245

RESUMO

BACKGROUND: Prediction of the extent of calcium supplement will facilitate safe and efficient management of hypocalcemia in the early postoperative stage of total parathyroidectomy with autotransplantation (PTXa) in patients with renal osteodystrophy. METHODS: The correlation between the extent of calcium deficiency, estimated by the amount of calcium supplement over 48 h after PTXa and using various parameters such as carboxy terminal parathyroid hormone (c-PTH), intact PTH (i-PTH), alkaline phosphatase (ALP), serum calcium, serum phosphorus, duration of hemodialysis, total weight of resected parathyroid glands and degree of subperiosteal resorption of the middle phalanx was examined in 49 patients who underwent PTX with subcutaneous autotransplantation. Bone mineral density (BMD) was also determined before, 3 months and 1 year after PTXa with dual energy X-ray absorptiometry (DEXA) in 13 patients. RESULTS: There was a positive correlation between pre-operative i-PTH level (r=0.56, P<0.0005) or ALP level (r=0.50, P<0.0005) and the amount of calcium supplement over 48 h after PTXa in these patients. Furthermore, the degree of subperiosteal resorption, determined by Jensen's classification, was significantly correlated with the amount of calcium supplement after PTX (P<0.05). Bone mineral density 3 months after (P<0.0005) and 1 year after PTXa (P<0.001) significantly increased compared with BMD before PTXa in all patients examined. CONCLUSION: These findings suggest that the pre-operative determination of i-PTH, ALP levels and degree of subperiosteal resorption allow the management of hypocalcemia safely and efficiently in renal osteodystrophy patients after PTXa.


Assuntos
Gluconato de Cálcio/uso terapêutico , Distúrbio Mineral e Ósseo na Doença Renal Crônica/cirurgia , Hipocalcemia/tratamento farmacológico , Paratireoidectomia , Complicações Pós-Operatórias/tratamento farmacológico , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
5.
J Vet Med Sci ; 61(3): 291-4, 1999 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10331206

RESUMO

The entire cDNA sequences were determined by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) techniques for equine copper/zinc superoxide dismutase (Cu/Zn-SOD) and manganese superoxide dismutase (Mn-SOD) through the use of total RNA extracted from the testis of an adult Thoroughbred. The results revealed a protein coding region for equine Cu/Zn-SOD with bases totaling 465 bp, accompanied by an estimated 154 residues of amino acids. As for equine Mn-SOD, its coding region contained a total of 669 bp and an estimated 222 residues of amino acids. Further, the expression of Cu/Zn-SOD and Mn-SOD genes were confirmed in the equine tissues by RT-PCR and in situ hybridization.


Assuntos
Regulação Enzimológica da Expressão Gênica , Cavalos/genética , Superóxido Dismutase/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA Complementar/química , Dados de Sequência Molecular
6.
Liver ; 19(6): 481-8, 1999 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-10661681

RESUMO

AIMS/BACKGROUND: We report in this study that, 10 mM nicotinamide can stimulate the proliferation of primary rat hepatocytes in serum-free Dulbecco's modified Eagle's medium supplemented with 10 ng/ml epidermal growth factor and that small hepatocyte colonies appear from 4 to 5 days after plating. We examined the effects of nicotinamide-related agents on the growth and differentiation of primary rat hepatocytes and on the appearance of small hepatocyte colonies. METHODS: As nicotinamide is an aqueous vitamin named niacin and known to act as an inhibitor of poly (ADP-ribose) polymerase (PARP), we therefore chose to examine the effects on hepatocytes of three nicotinamide-related agents, nicotinic acid (NA) which is also a niacin, 3-aminobenzamide (3-AB) which is a strong inhibitor of PARP but is not a niacin, and 3-acetylpyridine (3-AP) which is a weak inhibitor of PARP and also not a niacin. To examine their effects on the growth of the cells and on the formation of the colony, immunocytochemistry for BrdU was carried out. Expression of albumin, tryptophan 2,3-dioxygenase (TO), and connexin 32 (Cx32) mRNAs were used as marks of hepatic differentiation. Intracellular NAD+ content was also measured. RESULTS: At concentration of 10 mM, NA could not enhance the proliferation of mature hepatocytes but induced the appearance of small hepatocyte colonies. At concentration of 5 mM, 3-AB enhanced the proliferation of the hepatocytes but did not induce small hepatocyte colonies. On the other hand, although 10 mM 3-AP remarkably inhibited the DNA synthesis of the cells, the expression not only of albumin but also of TO and Cx32 mRNAs in the cells was well maintained for more than one week. The intracellular NAD+ concentration was correlated with the proliferation of the hepatocytes. CONCLUSION: These results suggest that the intracellular NAD+ content may be correlated with the proliferation of primary hepatocytes and that the supplementation of niacin in the medium may be important for the appearance of small hepatocyte colonies.


Assuntos
Benzamidas/farmacologia , Inibidores Enzimáticos/farmacologia , Fígado/citologia , Fígado/efeitos dos fármacos , Niacina/farmacologia , Inibidores de Poli(ADP-Ribose) Polimerases , Piridinas/farmacologia , Albuminas/metabolismo , Animais , Northern Blotting , Bromodesoxiuridina/metabolismo , Contagem de Células , Divisão Celular/efeitos dos fármacos , Células Cultivadas , Conexinas/metabolismo , Fígado/metabolismo , Masculino , NAD/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Triptofano Oxigenase/metabolismo , Proteína beta-1 de Junções Comunicantes
7.
Hepatology ; 29(1): 111-25, 1999 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-9862857

RESUMO

Hepatic cells isolated from an adult rat liver, consisting of small hepatocytes (SHs), mature hepatocytes (MHs), liver epithelial cells (LECs), Kupffer cells, sinusoidal endothelial cells, and stellate cells, were cultured in a medium supplemented with 10% fetal bovine serum, 10 mmol/L nicotinamide, 1 mmol/L ascorbic acid 2-phosphate, 10 ng/mL epidermal growth factor, and 1% dimethyl sulfoxide. The SHs rapidly proliferated and formed a colony. About 10% of cytokeratin 8 (CK8)-positive cells formed SH colonies. All SHs at day 10 immunocytochemically showed positivity for albumin, transferrin, CK8, and CK18, which are markers for hepatocytes. In contrast, alpha-fetoprotein (AFP)-, CK14-, OC2-, and glutathione S-transferase placental type (GST-P)-positive cells, which are thought to be markers for hepatic immature cells, were rarely observed. At day 20 some cells in the colonies were positive for AFP, CK7, CK19, and GST-P. LECs and stellate cells proliferated and surrounded the colonies. About 2 weeks after plating, piled up cells were often observed on the SH colonies. In those colonies LECs and stellate cells invaded under the colonies. The invasion of the cells and gradual deposits of extracellular matrix (ECM) such as type I collagen, type IV collagen, and laminin induced alteration of the shape of the SHs from relatively flat to cuboidal or rectangular. With the cellular structural changes, the expression of albumin, connexin 32 (Cx32), and tryptophan 2,3-dioxygenase (TO) messenger RNAs increased. In addition, overlapping nonparenchymal cells (NPCs) on the piled up cells induced the formation of duct- or cyst-like structures consisting of MHs. In the present experiment we showed that SHs could differentiate to MHs by interacting with NPCs and ECM. Thus, SHs may be "committed progenitor cells" that can further differentiate into MHs.


Assuntos
Fígado/ultraestrutura , Organoides/ultraestrutura , Animais , Bile/metabolismo , Canalículos Biliares/citologia , Northern Blotting , Western Blotting , Diferenciação Celular , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Matriz Extracelular/ultraestrutura , Fígado/citologia , Fígado/metabolismo , Masculino , Microscopia de Fluorescência , Organoides/metabolismo , Ratos , Ratos Sprague-Dawley
8.
J Gastroenterol Hepatol ; 13 Suppl: S70-7, 1998 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-9792037

RESUMO

Proliferation of adult rat hepatocytes is observed in serum-free Dulbecco's modified Eagle's medium (DMEM) supplemented with 10 mmol/L nicotinamide and 10 ng/mL epidermal growth factor (EGF). The proliferating cells are mainly mononucleate and form small cell colonies surrounded by mature hepatocytes. Although these cells in focal colonies have a less-differentiated appearance, immunocytochemically and ultrastructurally they possess hepatic characteristics. The size of small hepatocytes is one-third to half that of mature hepatocytes. Therefore, we call the cells forming a colony, small hepatocytes. The small hepatocytes can be subcultured for several passages. Furthermore, the cells are rich in the supernatant following 50 g centrifugation for 1 min after collagenase liver perfusion. When the cells are cultured in DMEM supplemented with 10% foetal bovine serum, 10 mmol/L nicotinamide, 1 mmol/L ascorbic acid 2-phosphate, 10 ng/mL EGF and 1% dimethyl sulphoxide, each small hepatocyte can clonally proliferate for more than 3 months. A small hepatocyte divides to form a colony and the number of cells reaches more than 100 within 20 days. With time in culture, cells with a large cytoplasm appear within a colony. They have many mitochondria and large peroxisomes with crystalline nucleoids and are typical, mature hepatocytes. Immunoreactivity to connexin 32 and well-developed bile canaliculus structures are often observed in the cell-cell borders. Thus, we suggest that small hepatocytes may be considered to be 'committed progenitor cells' that can further differentiate into mature hepatocytes.


Assuntos
Fígado/citologia , Animais , Diferenciação Celular , Células Cultivadas , Células Clonais , Fígado/ultraestrutura , Niacinamida/farmacologia , Ratos
9.
Aliment Pharmacol Ther ; 11(6): 1131-6, 1997 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-9663841

RESUMO

BACKGROUND: Primary and acquired resistance to antibiotics is an important factor in determining the reason for treatment failure in Helicobacter pylori infection. We examined the relationship between the susceptibility of H. pylori isolates and the efficacy of chemotherapy. METHODS: The minimal inhibitory concentrations (MICs) of metronidazole (MNZ), clarithromycin (CLAR) and amoxycillin (AMOX) of 320 H. pylori pre-treatment isolates were determined by the agar dilution method. In 290 patients with peptic ulcers. H. pylori infection was treated by dual or triple combination therapies for 2 weeks: one proton pump inhibitor (30 mg/day lansoprazole or 20 mg/day omeprazole) and one or two antibiotics (500 mg AMOX, 200 mg CLAR or 250 mg MNZ twice a day). MICs were also determined after the treatment failure. RESULTS: Among the drugs tested, for MNZ and CLAR, 8.1% and 9.1% of the isolates, respectively, were resistant, while no isolate was resistant to AMOX. After unsuccessful treatment using MNZ and CLAR, 66.7% and 70.61% of the isolates changed from sensitive to resistant, respectively. All isolates were sensitive to AMOX after treatment failure. CONCLUSIONS: The failure of the H. pylori treatment results in the induction of resistance to CLAR and/or MNZ. Regimens with a high cure rate should be used in order to prevent the generation of acquired resistance to antibiotics.


Assuntos
Quimioterapia Combinada/uso terapêutico , Inibidores Enzimáticos/uso terapêutico , Helicobacter pylori/isolamento & purificação , Inibidores da Bomba de Prótons , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Amoxicilina/uso terapêutico , Claritromicina/uso terapêutico , Suscetibilidade a Doenças , Resistência Microbiana a Medicamentos , Humanos , Japão , Metronidazol/uso terapêutico , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Resultado do Tratamento
10.
Metabolism ; 45(11): 1330-4, 1996 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8931635

RESUMO

Hypersecretion of prolactin (PRL) has been implicated as one of the factors that mediate ethanol-induced hypogonadism, but the site(s) in the central nervous system where ethanol acts to lead to the stimulation of PRL secretion is unknown. To clarify the site(s) of ethanol action, medial basal hypothalamic deafferentation (MBHD) or medial basal hypothalamic ablation (MBHA) were performed stereotaxically in male rats, and their PRL secretory capacity in response to acute ethanol administration was compared with that of intact or sham-operated controls. In intact control rats, plasma immunoreactive PRL concentration increased markedly (P < .001 v saline injection) following ethanol 400 to 500 mg/100 g body weight (BW) intraperitoneally (IP). The PRL response was dose-related and reached a maximum plateau level at 15 minutes. Plasma PRL returned to a near-basal level by 60 minutes. The response was blocked completely (P < .001) by pretreatment with dopamine (1 mg per rat), a specific inhibitor of adenohypophyseal PRL secretion. In sham-operated rats and in MBHD and MBHA rats, ethanol (500 mg/100 g BW IP) induced a significant (P < .001 to .05) elevation of PRL relative to the respective saline treatment. The basal level was significantly (P < .005) lower in the MBHD group (5.3 +/- 0.9 ng/mL) and significantly (P < .001) higher in the MBHA group (101.1 +/- 15.7 ng/mL) than in the sham group (17.2 +/- 5.9 ng/mL). These results suggest the following: (1) acute ethanol administration stimulates PRL secretion from the pituitary in a dose-related manner, (2) ethanol appears to have direct stimulatory effects on adenohypophyseal PRL secretion, and (3) extrahypothalamic brain areas exert a stimulatory influence and the hypothalamus an inhibitory influence on basal PRL secretion.


Assuntos
Etanol/farmacologia , Hipotálamo/efeitos dos fármacos , Prolactina/metabolismo , Animais , Dopamina/farmacologia , Feminino , Hipotálamo/fisiologia , Masculino , Prolactina/sangue , Ratos , Ratos Wistar , Fatores Sexuais
11.
Neurosci Lett ; 196(1-2): 85-8, 1995 Aug 18.
Artigo em Inglês | MEDLINE | ID: mdl-7501264

RESUMO

The protective effects of Rooibos tea (RT), Aspalathus linearis, against damage to the central nervous system (CNS) accompanying aging were examined by both the thiobarbituric acid reaction (TBA) and magnetic resonance imaging (MRI) methods in brains of chronically RT-treated rats. Ad libitum administration of RT was begun with 3-month-old Wistar female rats and continued for 21 months. The contents of TBA reactive substances (TBARS) in the frontal cortex, occipital cortex, hippocampus and cerebellum in 24-month-old rats after administration with water were significantly higher than those in young rats (5 weeks old). However, no significant increase of TBARS was observed in RT-administered aged rats. When MR images of the brains of 24-month-old rats with and without RT as well as 5-week-old rats were taken, a decrease of the signal intensity was observed in the cerebral cortex, hippocampus and cerebellum in MR images of aged rats without RT, whereas little change of the signal intensity was observed in MR images of the same regions of 24-month-old rats treated with RT, whose images were similar to those of young rats. These observations suggested that (1) the age-related accumulation of lipid peroxides in the brain was closely related to the morphological changes observed by MRI, and (2) chronic RT-administration prevented age-related accumulation of lipid peroxides in several regions of rat brain.


Assuntos
Envelhecimento/efeitos dos fármacos , Antioxidantes/farmacologia , Peróxidos Lipídicos/metabolismo , Chá , Administração Oral , Fatores Etários , Animais , Feminino , Imageamento por Ressonância Magnética , Ratos , Ratos Wistar , Tiobarbitúricos/farmacologia
12.
Plant Cell Physiol ; 36(1): 29-36, 1995 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-7719631

RESUMO

S-Adenosyl-L-methionine:scoulerine-9-O-methyltransferase (SMT) catalyzes the transfer of the S-methyl group of S-adenosyl-L-methionine to the 9-hydroxyl group of scoulerine during the biosynthesis of berberine. We have isolated functionally active cDNA clones (pCJSMTs) from a cDNA library prepared from cultured cells of Coptis japonica. The longest cDNA insert (pCJSMT1) had an open reading frame that encoded 351 amino acids, but the calculated molecular mass (38,364 Da) of the deduced product was slightly lower than the experimentally determined molecular mass of purified SMT. Rapid amplification of the 5' end of the cDNA indicated that the full-length cDNA of SMT consisted of 1,458 nucleotides that encoded 381 amino acids. When the full-length cDNA was expressed in E. coli, the molecular mass of the expressed SMT was greater than that of native SMT in Coptis cells. This result suggests that SMT might be produced in a pre-mature form and processed post-translationally. SMT was also found to exhibit sequence homology to other O-methyltransferases from plants and N-terminal region of the SMT polypeptide appeared to be necessary for enzymatic activity.


Assuntos
Metiltransferases/genética , Plantas Medicinais/enzimologia , Sequência de Aminoácidos , Sequência de Bases , Células Cultivadas , Clonagem Molecular , DNA Complementar , DNA de Plantas , Escherichia coli/genética , Dados de Sequência Molecular , Plantas Medicinais/citologia , Homologia de Sequência de Aminoácidos
13.
Eur J Biochem ; 225(1): 125-31, 1994 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-7925429

RESUMO

S-adenosyl-L-methionine:norcoclaurine 6-O-methyltransferase (norcoclaurine 6-O-methyltransferase), which catalyzes the transfer of the S-methyl group of S-adenosyl-L-methionine to the 6-hydroxyl group of 1,2,3,4-tetrahydro-1-[(4-hydroxyphenyl)methyl]-6,7- isoquinolinediol (norcoclaurine), was purified from cultured Coptis japonica cells and its enzymic properties were characterized. Purified norcoclaurine 6-O-methyltransferase had apparent pI 4.7, a native molecular mass of 95 kDa (determined by gel filtration) and subunit molecular mass of 40 kDa (SDS/PAGE). The enzyme did not require a divalent cation for activity, and the addition of Fe2+, Cu2+, Co2+, Zn2+, Mn2+, or Ni2+ at 5 mM severely inhibited enzyme activity. Neither p-chloromercuribenzoate, N-methylmaleimide nor iodoacetamide inhibited enzyme activity at 1 mM. 5,6-Dihydro-9,10-dimethoxybenzo[g]-1,3-benzodioxolo[5,6-a]qu inolizinium (berberine, the end-product of the biosynthetic pathway in which norcoclurine 6-O-methyltransferase catalyzes an intermediate step) also inhibited the activity by 50% at 10 mM. Norcoclaurine 6-O-methyltransferase methylated both (S)-norcoclaurine and (R)-norcoclaurine and (R,S)-norlaudanosoline. Further characterization of substrate-saturation kinetics and product inhibition of the purified enzyme indicated that norcoclaurine 6-O-methyltransferase follows a bi-bi ping-pong mechanism with Km values of 2.23 mM and 3.95 mM for (R,S)-norlaudanosoline and S-adenosyl-L-methionine, respectively, while Ki values for S-adenosylhomocysteine versus S-adenosyl-L-methionine and (R,S)-norlaudanosoline were 2.1 mM and 0.18 mM, respectively.


Assuntos
Metiltransferases/isolamento & purificação , Metiltransferases/metabolismo , Plantas Medicinais/enzimologia , Sequência de Aminoácidos , Células Cultivadas , Cromatografia , Cromatografia por Troca Iônica , Durapatita , Cinética , Metiltransferases/química , Dados de Sequência Molecular , Peso Molecular , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/isolamento & purificação , Especificidade por Substrato
14.
J Vet Med Sci ; 56(5): 933-8, 1994 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-7865597

RESUMO

Using a homemade MR imaging probe (Helmoholtz coil), MR images of brains of 5-week-old and 23- or 24-month-old Wistar rats were taken under a magnetic field of 7.05 T (Tesla). The probe was designed to fit the rat head and made by winding thin copper film round an acrylic tube with a 5-cm i.d., 10-cm length and 2-mm thickness. This was adjusted to resonate with the 300 MHz radiofrequency corresponding to the resonance frequency of 1H under a magnetic field of 7.05 T. MR images were obtained by T1-weighted and two-dimensional Fourier transformation techniques. The sagittal and coronal sections were imaged in 1-mm-thick slices. The size of the data matrix was 128 phase-encoded steps. Each image was obtained through eight acquisitions. A comparison of the MR images with those semi-microscopically taken at the same position of the coronal section revealed that the cerebral cortex, hippocampus, hypothalamus and thalamus were clearly imaged by this probe. With aging, MR images of cerebral cortices were observed with decreased signal intensities. Enlargement of the third ventricles and hypertrophy of cranical parietal bones were also recognized in sagittal MR images of aged rats. These observations were more marked in males than in females. From these observations it was concluded that this probe was applicable for MR imaging of rat brains under a magnetic field of 7.05 T.


Assuntos
Envelhecimento/fisiologia , Encéfalo/anatomia & histologia , Animais , Encéfalo/crescimento & desenvolvimento , Córtex Cerebral/anatomia & histologia , Córtex Cerebral/crescimento & desenvolvimento , Feminino , Hipocampo/anatomia & histologia , Hipocampo/crescimento & desenvolvimento , Hipotálamo/anatomia & histologia , Hipotálamo/crescimento & desenvolvimento , Imageamento por Ressonância Magnética/instrumentação , Imageamento por Ressonância Magnética/métodos , Masculino , Ratos , Ratos Wistar , Tálamo/anatomia & histologia , Tálamo/crescimento & desenvolvimento
15.
Biochem Biophys Res Commun ; 204(2): 578-84, 1994 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-7980517

RESUMO

Flavonoids are pigments of edible plants. We have recently reported that most flavonoids induce G1 arrest in human cancer cells, and that genistein (an isoflavone) specifically inhibits their cell cycle at G2-M phase. In the present study, apigenin (a flavone) was found to inhibit the proliferation of B104 rat neuronal cells, and flow-cytometric analysis showed that apigenin arrested their cell cycle at G2-M phase. This effect was dose-dependent and reversible when apigenin was removed from the culture medium. Microscopic observation showed that apigenin did not significantly increase the mitotic index compared with the control. Further, apigenin induced morphological differentiation, that is, elongation and arborization of neurites in B104 cells. This is the first report to show that apigenin inhibited the proliferation of malignant tumor cells by G2-M arrest and induced morphological differentiation.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Flavonoides/farmacologia , Fase G2/efeitos dos fármacos , Mitose/efeitos dos fármacos , Neurônios/citologia , Óleos Voláteis/farmacologia , Animais , Divisão Celular/efeitos dos fármacos , Linhagem Celular , Camomila , Índice Mitótico/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Plantas Medicinais , Ratos
16.
Biochim Biophys Acta ; 1156(2): 123-7, 1993 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-8381302

RESUMO

The relationship between the activity of pyrophosphate:fructose-6-phosphate 1-phosphotransferase (PFP) and the capacity for biosynthesis of macromolecules was examined in segments from different parts of hypocotyls of etiolated seedlings of Vigna mungo and V. radiata. The relative ratio of the maximum activity of PFP to that of ATP-dependent phosphofructokinase (PFK) (PFP/PFK ratio) was high in young tissues and decreased with differentiation and ageing of the tissues. The highest level of fructose-2,6-bisphosphate was observed in the youngest part of hypocotyls of V. mungo. The level was markedly decreased with ageing of tissues. The levels of PPi and ATP were also higher in younger parts than in older parts of the hypocotyls, but the ratio of the level of PPi to that of ATP was almost constant in all parts of the hypocotyl. A good correlation was found between the PFP/PFK ratio and the biosynthetic capacity, as estimated from the rate of incorporation of [U-14C]sucrose into ethanol-insoluble macromolecules.


Assuntos
Fabaceae/enzimologia , Fosfotransferases/metabolismo , Plantas Medicinais , Envelhecimento , Diferenciação Celular , Frutosedifosfatos/biossíntese , Fosfofrutoquinase-1/metabolismo
17.
Arteriosclerosis ; 10(5): 732-7, 1990.
Artigo em Inglês | MEDLINE | ID: mdl-2403301

RESUMO

The effect of dietary magnesium (Mg) on the development of atherosclerosis in cholesterol-fed rabbits was investigated. Male New Zealand White rabbits (n = 31) were placed on five kinds of diets: regular, 1% cholesterol, and 1% cholesterol diets supplemented with either 300, 600, or 900 mg (as Mg) of Mg sulfate. The regular and 1% cholesterol diets contained 400 mg of Mg per 100 g. Each rabbit received 100 g daily of the appropriate diet. Additional Mg was well tolerated and did not affect blood pressure or body weight. The rabbits were sacrificed after 10 weeks, and the oil red O-positive atherosclerotic area that covered the aortic intima and the cholesterol content of the aorta was measured. Additional Mg decreased both the area of the aortic lesions and the cholesterol content of the aortas in a dose-dependent manner. The 1% cholesterol diet significantly increased plasma cholesterol and triglyceride concentrations and decreased high density lipoprotein (HDL) cholesterol concentration. Additional Mg had no further effect on cholesterol and HDL cholesterol concentrations, but it slightly decreased the rise in triglyceride concentration. These results indicate that dietary Mg prevents the development of atherosclerosis in cholesterol-fed rabbits by inhibiting lipid accumulation in the aortic wall.


Assuntos
Arteriosclerose/sangue , Magnésio/farmacologia , Animais , Aorta/metabolismo , Aorta/patologia , Doenças da Aorta/patologia , Arteriosclerose/induzido quimicamente , Arteriosclerose/patologia , Análise Química do Sangue , Pressão Sanguínea , Peso Corporal , Cálcio/metabolismo , Colesterol/metabolismo , Colesterol na Dieta , Dieta , Magnésio/administração & dosagem , Magnésio/metabolismo , Masculino , Coelhos
18.
Jpn J Antibiot ; 39(3): 815-22, 1986 Mar.
Artigo em Japonês | MEDLINE | ID: mdl-3016359

RESUMO

SMANCS is a conjugate protein of copolymer of styrene-maleic acid [SMA] (molecular weight: 1,500) and an antitumor protein neocarzinostatin [NCS] (molecular weight: 11,700). It has an approximate molecular weight of 15,000. We report here stability of SMANCS in oil and in water, and NCS in water, under various physical conditions such as exposure to heat, UV, pH, and ultrasonic treatment. Then, we carried out an experiment of transfer of SMANCS in lipid contrast medium [lipiodol] (oil phase) to water phase (blood and physiological saline) in vitro. Results are summarized as follows: In aqueous condition, SMANCS is far more stable than NCS against the exposure to heat and UV, though it is inactivated by excessive exposures. SMANCS in an oily medium was found much more stable even at higher temperatures than in the aqueous phase. Both SMANCS and NCS are the most stable at pH 4.9-6.0. SMANCS dissolved in oil transferred to water phase slowly, having T1/10 of 24 hours (in case of lipiodol). This helps maintaining the anticancer effect of the drug in vivo for a long period of time. SMANCS in lipiodol was found to exert its action against cultured tumor cells as in an aqueous solution.


Assuntos
Antibióticos Antineoplásicos , Antineoplásicos , Furanos , Anidridos Maleicos , Poliestirenos , Zinostatina , Antibióticos Antineoplásicos/efeitos da radiação , Antineoplásicos/efeitos da radiação , Sangue , Estabilidade de Medicamentos , Furanos/efeitos da radiação , Temperatura Alta , Concentração de Íons de Hidrogênio , Óleo Iodado , Anidridos Maleicos/efeitos da radiação , Poliestirenos/efeitos da radiação , Cloreto de Sódio , Soluções , Ultrassom , Raios Ultravioleta , Água , Zinostatina/análogos & derivados , Zinostatina/efeitos da radiação
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