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1.
J Obstet Gynaecol ; 37(4): 487-491, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28421907

RESUMO

Laparoscopic entry techniques vary amongst surgeons and gynaecologists, with gynaecologists favouring Veress needle entry. Recent RCOG/BSGE recommendations have recommended retrieval of ovarian masses via the umbilical port with resultant less postoperative pain and a faster retrieval time than with retrieval through lateral ports of the same size. This is a prospective observational study reviewing the Hasson entry technique and the introduction of retrieval of specimens via the umbilicus in patients scheduled for a laparoscopy procedure at our day surgery unit. We found no immediate or major surgical complications and all specimens were successfully retrieved through the umbilicus. Pain scores were low. We recommend the technique for extirpative gynaecological surgery. Impact Statement This study confirms that conversion from the closed Veress to the open Hasson technique is achievable in a university hospital setting. Operator confidence during the learning phase is enhanced by the use of ultrasound to locate and measure the depth of the umbilical ligament. Retrieval of benign adnexal specimens through the umbilicus was very satisfactory.


Assuntos
Doenças dos Anexos/cirurgia , Procedimentos Cirúrgicos em Ginecologia/métodos , Laparoscopia/métodos , Umbigo/cirurgia , Adolescente , Adulto , Idoso , Feminino , Humanos , Laparoscopia/efeitos adversos , Pessoa de Meia-Idade , Neoplasias Ovarianas/cirurgia , Dor Pós-Operatória/etiologia , Estudos Retrospectivos , Ultrassonografia , Umbigo/diagnóstico por imagem , Adulto Jovem
2.
Diabetes Obes Metab ; 18(6): 628-32, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-26889911

RESUMO

The aim of the present study was to assess the hypoglycaemia risk and safety of dapagliflozin compared with sulphonylurea during the fasting month of Ramadan. In this 12-week, randomized, open-label, two-arm parallel group study, 110 patients with type 2 diabetes who were receiving sulphonylurea and metformin were randomized either to receive 10 mg (n = 58) of dapagliflozin daily or to continue receiving sulphonylurea (n = 52). The primary outcome was to compare the effects of dapagliflozin and sulphonylurea on the proportions of patients with at least one episode of hypoglycaemia during Ramadan, as well as to assess the safety of dapagliflozin when used to treat patients observing Ramadan. A lower proportion of patients had reported or documented hypoglycaemia in the dapagliflozin group than in the sulphonylurea group: 4 (6.9%) versus 15 (28.8%); p = 0.002. The relative risk of any reported or documented hypoglycaemia in the 4th week of Ramadan was significantly lower in the dapagliflozin group: RR=0.24, 95%CI: 0.09, 0.68; p=0.002. No significance differences were observed between the two groups regarding postural hypotension (13.8 vs 3.8%; p = 0.210) or urinary tract infections (10.3 vs 3.8%; p = 0.277). In conclusion, fewer patients exhibited hypoglycaemia in the dapagliflozin group than in the sulphonylurea group.


Assuntos
Compostos Benzidrílicos/administração & dosagem , Diabetes Mellitus Tipo 2/tratamento farmacológico , Substituição de Medicamentos/métodos , Jejum , Glucosídeos/administração & dosagem , Hipoglicemia/prevenção & controle , Hipoglicemiantes/administração & dosagem , Islamismo , Metformina/administração & dosagem , Compostos de Sulfonilureia/administração & dosagem , Adulto , Idoso , Compostos Benzidrílicos/efeitos adversos , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/complicações , Quimioterapia Combinada , Jejum/sangue , Feminino , Glucosídeos/efeitos adversos , Humanos , Hipoglicemia/induzido quimicamente , Hipoglicemiantes/efeitos adversos , Masculino , Metformina/efeitos adversos , Pessoa de Meia-Idade , Inibidores do Transportador 2 de Sódio-Glicose , Compostos de Sulfonilureia/efeitos adversos , Adulto Jovem
3.
Eur J Gynaecol Oncol ; 37(5): 632-637, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-29787000

RESUMO

Secondary healing of complicated vulvar and groin wounds is a major challenge due to its moist condition and at risk of contamination by colonic flora. Vacuum assisted closure is the controlled application of sub-atmospheric pressure to the local wound environment using a sealed dressing connected to a vacuum pump. MATERIALS AND METHODS: The NPWT consists of an open-pore polyurethane ether foam sponge, an adhesive cover, fluid collection system, and suction pump that generates negative pressure. Direct application of sponge to blood vessels, bone, nerves or intact skin is avoided. The dressing and tubing are changed every 48-72 hours. RESULTS: Eight patients had NPWT following the vulva and/or groin surgery. Pain during removal of the sponge was the main adverse event requiring narcotic analgesia. All wounds healed completely. One patient is dead of disease progression. Others are alive without disease at four to 48 months. CONCLUSION: Wound breakdown in vulvar and groin surgery is an infrequent occurrence because of the rarity of full radical excision for vulva cancer and infralevator pelvic exenterative surgery. The present experience with NPWT was favourable.


Assuntos
Virilha/cirurgia , Tratamento de Ferimentos com Pressão Negativa/métodos , Exenteração Pélvica , Vulva/cirurgia , Neoplasias Vulvares/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Pessoa de Meia-Idade , Neoplasias Vulvares/fisiopatologia , Cicatrização
4.
Eur J Gynaecol Oncol ; 35(2): 143-8, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24772916

RESUMO

UNLABELLED: There is considerable variation within and between cancer centers in the practice of appendectomy as part of cytoreductive surgery for ovarian carcinoma and in the surgical staging of endometrial carcinoma. The purpose of this study was to determine the prevalence and the type of appendiceal pathology, the morbidity associated with appendectomy in gynaecologic cancer surgery. MATERIALS AND METHODS: This is a retrospective review of all cytoreductive surgery for ovarian carcinoma and surgical staging for endometrial carcinoma with appendectomy over a four year period. RESULTS: Two hundred and fifty-one patients (38 patients for endometrial carcinoma surgery and 213 patients for ovarian cytoreduction) had an appendectomy performed. Metastases to the appendix was present in 46 (23.2%) of primary ovarian carcinoma and one (2.6%) primary endometrial carcinosarcoma. The appendix was more likely to be involved in advanced stage ovarian cancer with positive peritoneal washings, omental deposits, grade 3 differentiation, and papillary serous histology. Sixteen (6.4%) co-incidental primary appendiceal tumours were detected. No postoperative morbidity specific to appendectomy was identified. One case of ovarian carcinoma was upstaged from IC to IIIA by the appendiceal metastases. There was no upstaging of disease in the endometrial carcinoma group. DISCUSSION: Appendectomy is an integral part of ovarian cytoreductive surgery but the authors found it did not upstage the disease in a clinically significant manner. The incidence of co-incidental appendiceal primary tumours was high in this series and may add value to the procedure in preventing further surgeries. The absence of procedure related morbidity is reassuring. The authors recommend appendectomy for all ovarian staging surgery and its consideration in type 2 endometrial cancer.


Assuntos
Adenocarcinoma/secundário , Neoplasias do Apêndice/secundário , Apêndice/patologia , Neoplasias do Endométrio/patologia , Neoplasias Ovarianas/patologia , Adenocarcinoma/cirurgia , Adenocarcinoma Mucinoso/secundário , Adenocarcinoma Mucinoso/cirurgia , Adenocarcinoma Papilar/secundário , Adenocarcinoma Papilar/cirurgia , Idoso , Idoso de 80 Anos ou mais , Apendicectomia , Neoplasias do Apêndice/patologia , Neoplasias do Apêndice/cirurgia , Apêndice/cirurgia , Tumor Carcinoide/patologia , Carcinoma Endometrioide/secundário , Carcinoma Endometrioide/cirurgia , Carcinoma de Células em Anel de Sinete/patologia , Cistadenoma Mucinoso/patologia , Neoplasias do Endométrio/cirurgia , Feminino , Humanos , Pessoa de Meia-Idade , Mixoma/patologia , Neoplasias Primárias Múltiplas/patologia , Neoplasias Ovarianas/cirurgia , Estudos Retrospectivos
5.
Nutr Diabetes ; 4: e107, 2014 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-24535618

RESUMO

Post-gestational diabetes mellitus (GDM) women are recommended weight loss to manage increased cardio-metabolic risks. We investigated the effects of lowering diet glycaemic index (GI) on fasting blood glucose (FBG), serum lipids, body weight and composition of post-GDM women with varying fasting insulin levels (INS). Seventy-seven Asian, non-diabetic women with previous GDM (aged 20-40 years, mean BMI: 26.4±4.6 kg m(-2)) were recruited. At baseline, 20 subjects with INS <2 µIU ml(-1) and 18 with INS 2 µIU ml(-1) received conventional dietary recommendations (CHDR) only. CHDR emphasised energy and fat intake restriction and encouraged increase in dietary fibre intakes. Twenty-four subjects with INS <2 µIU ml(-1) and 15 with INS 2 µIU ml(-1), in addition to CHDR, received low-GI education (LGI). Changes in FBG, serum lipids, body weight and body composition were evaluated. Subjects with INS <2 µIU ml(-1) had similar outcomes with both diets. After 1 year, subjects with INS 2 µIU ml(-1) who received LGI education had reductions in FBG and triglycerides. Subjects who received CHDR observed increase in both FBG and triglycerides (P<0.05). Among all subjects, diet GI was lower and dietary fibre intakes were higher in LGI compared with CHDR subjects (all P<0.05). Thus, in Asian post-GDM women with normal/higher INS, adding low-GI education to CHDR improved management of FBG and triglycerides.

6.
Eur J Obstet Gynecol Reprod Biol ; 170(1): 251-4, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23880597

RESUMO

OBJECTIVE: Salpingectomy is proposed as a prophylactic measure to reduce the incidence of tubo-ovarian/pelvic serous cancers. We surveyed the attitudes of obstetrician/gynecologists to incorporating salpingectomy opportunistically into surgery for benign conditions, and electively for young BRCA mutation carriers. STUDY DESIGN: A questionnaire, designed to assess current standard clinical practice and willingness to perform salpingectomy for female sterilization at abdominal hysterectomy for benign disease (ABH), vaginal benign hysterectomy (VBH) and electively for women with BRCA mutations who wish to postpone oophorectomy was mailed to obstetrician/gynecologists working in Irish hospitals. RESULTS: In their current practice of interval female sterilization 96% of gynecologists applied clips at laparoscopy and 4% performed salpingectomy, and 73% were willing to consider salpingectomy. Eighty-one percent were willing to consider salpingectomy for sterilization at cesarean section. Gynecologists performing hysterectomy (without oophorectomy) for benign conditions did salpingectomy in 26% at ABH and 5.4% at VBH, and now 90% would consider salpingectomy at ABH and 66% at VBH. Two-thirds of respondents would consider salpingectomy for women at genetic risk of ovarian cancer who want to postpone oophorectomy. CONCLUSION: Changing general gynecological practice to include more opportunistic salpingectomy has the potential to reduce the incidence of serous cancers. The majority of gynecologists are willing to incorporate more salpingectomies into their surgical practices and consider elective salpingectomy as an interim measure for women with defined genetic risk of pelvic serous cancer.


Assuntos
Atitude do Pessoal de Saúde , Cistadenocarcinoma Seroso/prevenção & controle , Neoplasias dos Genitais Femininos/prevenção & controle , Salpingectomia/psicologia , Feminino , Humanos
7.
Phys Rev Lett ; 108(2): 020603, 2012 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-22324666

RESUMO

We demonstrate theoretically and experimentally the phenomenon of vibrational resonance in a periodic potential, using cold atoms in an optical lattice as a model system. A high-frequency (HF) drive, with a frequency much larger than any characteristic frequency of the system, is applied by phase modulating one of the lattice beams. We show that the HF drive leads to the renormalization of the potential. We used transport measurements as a probe of the potential renormalization. The very same experiments also demonstrate that transport can be controlled by the HF drive via potential renormalization.


Assuntos
Luz , Modelos Teóricos , Fenômenos Ópticos , Transporte Biológico , Vibração
8.
Phys Rev E Stat Nonlin Soft Matter Phys ; 84(2 Pt 1): 021127, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21928969

RESUMO

Motivated by recent work [D. Cubero et al., Phys. Rev. E 82, 041116 (2010)], we examine the mechanisms which determine current reversals in rocking ratchets as observed when varying the frequency of the drive. We found that a class of these current reversals in the frequency domain is precisely determined by dissipation-induced symmetry breaking. Our experimental and theoretical work thus extends and generalizes the previously identified relationship between dynamical and symmetry-breaking mechanisms in the generation of current reversals.

9.
J Obstet Gynaecol ; 30(7): 675-8, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20925608

RESUMO

This study was undertaken to determine the presence of thyroid autoantibodies and associated pregnancy complications from 49 pregnant women with thyroid disease. There were 31 (63%) women with Graves' disease (GD) and 18 (37%) with primary hypothyroidism (PHT). A total of 26 (53.1%) women, 19 (61%) with GD and seven (39%) with PHT, had positive antibodies. Six had thyroid peroxidase antibodies (TPO), one with thyroglobulin antibody (TG) and eight had TSH receptor antibodies (TR). Two had a mixture of antibodies involving TG/TPO (one GD vs one PHT), four with TG/TPO/TR (all had GD) and five with TPO/TR (four with GD vs one with PHT). There were associations in women with positive thyroid antibodies and pre-eclampsia (15.4%), abruptio placenta (4%), caesarean deliveries (31%), postpartum thyroiditis (19.2%) and abnormal neonatal thyroid function (15.4%). Women with positive thyroid antibodies in pregnancy need close care during and after pregnancy, as they can develop complications affecting both mother and fetus.


Assuntos
Autoanticorpos/sangue , Hipertireoidismo/imunologia , Hipotireoidismo/imunologia , Imunoglobulinas Estimuladoras da Glândula Tireoide/sangue , Complicações na Gravidez/imunologia , Descolamento Prematuro da Placenta/epidemiologia , Descolamento Prematuro da Placenta/imunologia , Adulto , Cesárea/estatística & dados numéricos , Feminino , Humanos , Hipertireoidismo/epidemiologia , Hipotireoidismo/epidemiologia , Iodeto Peroxidase/imunologia , Pré-Eclâmpsia/epidemiologia , Pré-Eclâmpsia/imunologia , Gravidez , Complicações na Gravidez/epidemiologia , Receptores da Tireotropina/imunologia , Estudos Soroepidemiológicos , Tireoglobulina/imunologia , Tireoidite/epidemiologia , Tireoidite/imunologia , Adulto Jovem
10.
Singapore Med J ; 50(11): 1077-9, 2009 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-19960163

RESUMO

INTRODUCTION: The present study aimed to obtain preliminary tympanometric data of young Malay adults and to compare the results between genders. METHODS: 96 undergraduate students (49 males and 47 females), aged 19-25 (mean and standard deviation 21.14 +/- 1.31) years, participated in this study. Otoscopic examination, pure tone audiometry, qualitative tympanogram and ipsilateral acoustic reflex were measured to ensure a clear ear canal, normal hearing and normal middle ear function, prior to tympanometric measurement. As a result, a total of 154 ears (80 ears from males and 74 ears from females) were selected for further statistical analyses. The tympanometric parameters measured were peak compensated static acoustic admittance (Peak Y(tm)), tympanometric width (TW) and equivalent ear canal volume (V(ea)). RESULTS: The results showed that the mean Peak Y(tm), V(ea) and TW for males were 0.81 mmhos, 1.48 cubic cm and 113.67 daPa, respectively. The mean Peak Y(tm), V(ea) and TW for females were 0.63 mmhos, 1.12 cubic cm and 98.04 daPa, respectively. Males were found to have significantly higher mean V(ea) and mean Peak Y(tm) than females. However no significant gender difference was observed in the mean TW. CONCLUSION: The current study suggests that young Malay adults may require gender-specific Peak Y(tm) and V(ea) values when implementing a quantitative approach in tympanogram interpretation.


Assuntos
Testes de Impedância Acústica/métodos , Acústica , Audiometria/métodos , Audiometria de Tons Puros , Meato Acústico Externo/patologia , Etnicidade , Feminino , Humanos , Malásia , Masculino , Otoscopia/métodos , Valores de Referência , Fatores Sexuais , Adulto Jovem
11.
Med J Malaysia ; 61(5): 641-3, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17623971

RESUMO

We report a case of an adult who presented with progressive swelling in the right eye with suggestive of intracranial lesion on imaging. Histopathological revealed a lacrimal gland malignant mixed tumour.


Assuntos
Neoplasias Encefálicas/cirurgia , Neoplasias Oculares/cirurgia , Doenças do Aparelho Lacrimal/cirurgia , Aparelho Lacrimal/patologia , Adulto , Neoplasias Encefálicas/diagnóstico , Neoplasias Encefálicas/secundário , Neoplasias Oculares/diagnóstico , Neoplasias Oculares/patologia , Evolução Fatal , Humanos , Doenças do Aparelho Lacrimal/diagnóstico , Doenças do Aparelho Lacrimal/patologia , Masculino
12.
Diabetologia ; 48(12): 2650-60, 2005 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-16270194

RESUMO

AIMS/HYPOTHESIS: We quantified the glomerular expression of thrombospondin-1 (THBS1, also known as TSP-1), transforming growth factor beta 1 (TGFB1, also known as TGF-beta1) and connective tissue growth factor (CTGF) at each stage of diabetic nephropathy. We also examined the roles of THBS1 and CTGF in mediating high-glucose- and glycated-albumin-induced synthesis of the matrix protein, fibronectin, by mesangial cells. METHODS: THBS1, latent and active TGFB1, and CTGF, were detected by immunohistochemistry and in situ hybridisation in biopsies from 19 insulin-dependent diabetic patients with incipient, manifest and advanced diabetic nephropathy, and in 11 control kidneys. Findings were quantified by image analysis. Human mesangial cells were cultured with normal or high glucose, albumin or glycated albumin (Amadori product), +/-THBS1 or CTGF antisense oligonucleotides, or with peptide W, an inhibitor of TGFB1 bioactivation by THBS1. Proteins were measured by western blot analysis or ELISA. RESULTS: In glomeruli of normal kidneys, mRNA and protein levels for THBS1, latent-TGFB1 and CTGF were low. They were increased in the incipient stage of diabetic nephropathy, predominantly in mesangial areas, with further increases at later stages of the disease. Little or no active TGFB1 immunostaining was detected prior to manifest diabetic nephropathy. In contrast to high-glucose conditions, increases in fibronectin synthesis that were stimulated by glycated albumin were not dependent on THBS1 activation of latent TGFB1. However, increased fibronectin synthesis in both conditions required CTGF. CONCLUSIONS/INTERPRETATION: Increased glomerular expression of all three factors occurs from the earliest stage of diabetic nephropathy. In contrast to THBS1, CTGF is required for mesangial synthesis of fibronectin stimulated by high glucose or glycated albumin, and is thus a potential therapeutic target.


Assuntos
Nefropatias Diabéticas/metabolismo , Mesângio Glomerular/metabolismo , Hiperglicemia/metabolismo , Proteínas Imediatamente Precoces/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Trombospondina 1/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Adulto , Idoso , Western Blotting , Estudos de Casos e Controles , Fator de Crescimento do Tecido Conjuntivo , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/metabolismo , Nefropatias Diabéticas/genética , Nefropatias Diabéticas/patologia , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Feminino , Fibronectinas/genética , Fibronectinas/metabolismo , Regulação da Expressão Gênica , Mesângio Glomerular/patologia , Glucose/farmacologia , Glucose/fisiologia , Produtos Finais de Glicação Avançada , Humanos , Proteínas Imediatamente Precoces/genética , Imuno-Histoquímica , Peptídeos e Proteínas de Sinalização Intercelular/genética , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/análise , RNA Mensageiro/genética , Albumina Sérica/farmacologia , Trombospondina 1/genética , Fator de Crescimento Transformador beta/genética , Albumina Sérica Glicada
13.
Med J Malaysia ; 59(4): 495-501, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15779582

RESUMO

The albumin globulin ratio (A/G ratio) is meant to represent the ratio of alterations in serum proteins, since, in liver disease, globulins (G) rise following serum albumin (SA) decrease. pathophysiological value, its' use has been limited. Alternatively, we have developed an index, the globulin compensation index (GCI) to measure the changes in serum globulins when albumin is decreased. The index is calculated as follows: G - 25 / 35 - SA. The GCI has been tested using retrospective patients' data from the Hospital Universiti Sains Malaysia. Analysis of the data shows that the GCI may be of potential value in showing the actual serum protein status, especially in cases where globulins are decreased along with albumin. Furthermore, globulin rise in cases with reduced albumin was found in 72.3% of cases of hepatic diseases, whereas this finding occurred in up to 32.3% of cases of non-hepatic, systemic diseases.


Assuntos
Albumina Sérica/análise , Soroglobulinas/análise , Humanos , Nefropatias/sangue , Estudos Retrospectivos
14.
Malays J Med Sci ; 10(1): 46-51, 2003 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23365500

RESUMO

The utilization of the chemical laboratory resources at the Hospital Sains Malaysia was evaluated. More than 100,000 test requests received and performed over a 12-month period, were analyzed retrospectively. The analysis conducted included the abnormal results obtained, the degree of duplication of tests, and the extent of test-panel ordering. It was found that a relatively moderate degree of over-ordering was evident. The findings suggested that the main reasons for over-ordering were the use of panel tests of ordering, in addition to a small, yet significant degree of duplication. Strategies for cutting down the test ordering have been reviewed and discussed.

15.
Kidney Int ; 60(5): 1756-64, 2001 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11703593

RESUMO

BACKGROUND: Accumulation of mesangial extracellular matrix is a major characteristic of diabetic nephropathy (DN). Expression of several extracellular matrix proteins is up-regulated in human mesangial cells (HMC) cultured in high glucose. One protein, fibronectin (FN), associates to form an insoluble disulfide-linked matrix and possesses inherent protein-disulfide isomerase (PDI) activity. Bacitracin is a known PDI inhibitor. We tested the hypothesis that inhibiting FN-PDI activity with bacitracin would disrupt excessive FN-matrix assembly by cultured HMCs grown under high glucose conditions. METHODS: The effect of bacitracin on FN-PDI activity was tested using an RNase-refolding assay. High glucose cultures of HMC were labeled with (3)H-leucine, with and without bacitracin, and (3)H-FN immunoprecipitated from the medium and sequential extracts of cell layers to distinguish insoluble FN. FN transcription was assessed by reverse transcription-polymerase chain reaction (RT-PCR). Pericellular FN-matrix was examined by immunohistology. RESULTS: Bacitracin inhibited the PDI activity of FN, with maximal inhibition at 1.0 mmol/L. Treatment of HMC cultures grown in high glucose with bacitracin brought about changes in the distribution of newly synthesized FN. With increasing concentrations of bacitracin there was a significant reduction in the level of FN present as an insoluble matrix of HMC cultures maintained in high glucose, and a corresponding increase in FN in medium. Decreases in FN matrix laid down by HMCs treated with different concentrations of bacitracin were seen by immunohistology. FN mRNA levels were unchanged. CONCLUSION: PDI inhibition of FN reduces its association into an insoluble matrix and potentially provides a new approach to reduce excessive matrix deposition in DN.


Assuntos
Antibacterianos/farmacologia , Fibronectinas/química , Mesângio Glomerular/química , Glucose/farmacologia , Macrolídeos , Polienos/farmacologia , Células Cultivadas , Fibronectinas/genética , Humanos , Isomerases de Dissulfetos de Proteínas/antagonistas & inibidores , RNA Mensageiro/análise
16.
Biochem J ; 359(Pt 1): 89-97, 2001 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-11563972

RESUMO

Connective tissue growth factor (CTGF) is a secreted cysteine-rich protein now considered as an important effector molecule in both physiological and pathological processes. An increasing amount of evidence indicates that CTGF plays a key role in the pathogenesis of different fibrotic disorders including diabetic nephropathy. However, the molecular mechanisms by which CTGF exerts its effects are not known. Here we provide the first evidence for the existence of an intracellular transport pathway for the growth factor in human mesangial cells. Our results demonstrate that CTGF is internalized from the cell surface in endosomes and accumulates in a juxtanuclear organelle from which the growth factor is then translocated into the cytosol. In the cytosol CTGF is phosphorylated by protein kinase C and PMA treatment can enhance this phosphorylation. Phosphorylated CTGF may have an important role in the cytosol, but it is also translocated into the nucleus where it may directly affect transcription.


Assuntos
Mesângio Glomerular/metabolismo , Proteínas Imediatamente Precoces/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Transporte Biológico , Western Blotting , Carcinógenos/farmacologia , Células Cultivadas , Fator de Crescimento do Tecido Conjuntivo , Endocitose , Técnica Indireta de Fluorescência para Anticorpo , Mesângio Glomerular/citologia , Humanos , Fosforilação , Testes de Precipitina , Proteína Quinase C/metabolismo , Transporte Proteico , RNA Mensageiro/metabolismo , Frações Subcelulares , Acetato de Tetradecanoilforbol/farmacologia , Transfecção , Fator de Crescimento Transformador beta/metabolismo
17.
Biochem J ; 359(Pt 1): 77-87, 2001 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-11563971

RESUMO

We characterized a rabbit polyclonal antibody raised against human recombinant connective tissue growth factor (CTGF). The antibody recognised a higher molecular mass form (approx. 56 kDa) of CTGF in mesangial cell lysates as well as the monomeric (36-38 kDa) and lower molecular mass forms (<30 kDa) reported previously. Immunohistochemistry detected CTGF protein in glomeruli of kidneys of non-obese diabetic mice 14 days after the onset of diabetes, and this was prominent by 70 days. CTGF protein is also present in glomeruli of human patients with diabetic nephropathy. No CTGF was detected in either normal murine or human glomeruli. Transient transfection of a transformed human mesangial cell line with a CTGF-V5 epitope fusion protein markedly increased fibronectin and plasminogen activator inhibitor-1 synthesis in cultures maintained in normal glucose (4 mM) conditions; a CTGF-antisense construct reduced the elevated synthesis of these proteins in high glucose (30 mM) cultures. Culture of primary human mesangial cells for 14 days in high glucose, or in low glucose supplemented with recombinant CTGF or transforming growth factor beta1, markedly increased CTGF mRNA levels and fibronectin synthesis. However, whilst co-culture with a CTGF-antisense oligonucleotide reduced the CTGF mRNA pool by greater than 90% in high glucose, it only partially reduced fibronectin mRNA levels and synthesis. A chick anti-CTGF neutralizing antibody had a similar effect on fibronectin synthesis. Thus both CTGF and CTGF-independent pathways mediate increased fibronectin synthesis in high glucose. Nevertheless CTGF expression in diabetic kidneys is likely to be a key event in the development of glomerulosclerosis by affecting both matrix synthesis and, potentially through plasminogen activator inhibitor-1, its turnover.


Assuntos
Diabetes Mellitus Experimental/etiologia , Nefropatias Diabéticas/etiologia , Proteínas Imediatamente Precoces/fisiologia , Peptídeos e Proteínas de Sinalização Intercelular/fisiologia , Animais , Biópsia , Western Blotting , Células Cultivadas , Clonagem Molecular , Fator de Crescimento do Tecido Conjuntivo , DNA Complementar , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patologia , Nefropatias Diabéticas/metabolismo , Nefropatias Diabéticas/patologia , Ensaio de Imunoadsorção Enzimática , Fibronectinas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Mesângio Glomerular/metabolismo , Mesângio Glomerular/patologia , Glucose/metabolismo , Glucose/farmacologia , Humanos , Técnicas Imunoenzimáticas , Imunoglobulina G/farmacologia , Rim/metabolismo , Camundongos , Camundongos Endogâmicos NOD , Oligonucleotídeos Antissenso/farmacologia , Inibidor 1 de Ativador de Plasminogênio/metabolismo , RNA Mensageiro , Proteínas Recombinantes de Fusão/metabolismo , Transfecção , Fator de Crescimento Transformador beta/metabolismo , Fator de Crescimento Transformador beta1
18.
Int J Mol Med ; 8(2): 193-8, 2001 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11445874

RESUMO

Amplification of int-2/FGF-3 gene was investigated by differential polymerase chain reaction (dPCR) in 440 archival primary breast carcinoma tissues. Of these, 23 were comedo ductal carcinoma in situ (DCIS), 18 were non-comedo DCIS, 41 were comedo DCIS with adjacent invasive ductal carcinomas, 19 were non-comedo DCIS with adjacent invasive ductal carcinomas, 270 were invasive ductal carcinomas, 33 were invasive lobular carcinomas, 21 were colloid carcinomas and 15 were medullary carcinomas. Int-2 was amplified in 22% (96/440) of the primary breast carcinomas. It was shown that int-2 was amplified in 13% (3/23) of the comedo DCIS, 17% (7/41) of the comedo DCIS and 29% (12/41) of the adjacent invasive ductal carcinomas, 26% (71/270) of the invasive ductal carcinomas, 18% (6/33) of the invasive lobular carcinomas, 10% (2/21) of the colloid carcinomas and 13% (2/15) of the medullary carcinomas. In contrast, int-2 was not amplified in non-comedo DCIS and invasive ductal carcinomas with adjacent non-comedo DCIS lesions. A significant association was observed between int-2 amplification in the in situ components and adjacent invasive lesion (P<0.05). All tumors with int-2 amplification in the in situ lesions (7/7) also demonstrated same degree of amplification in the adjacent invasive components. However, 9% (5/53) of the tumors with no amplified int-2 gene in the in situ components showed int-2 amplification in the adjacent invasive lesions. A significant relationship was noted between amplification of int-2 and lymph node metastases (P<0.05) and poorly differentiated tumors (P<0.05) but not with estrogen receptor status (P>0.05) and proliferation index (Ki-67 and PCNA) (P>0.05). In Malaysia, majority of the patients belong to younger age group (<50 years old) but a comparison of the age groups showed that the amplification of int-2 was not statistically associated with patient age (P>0.05). These observations indicate that amplification of int-2 tends to strengthen the view that int-2 may have the potential to be an indicator of poor prognosis regardless of the age of the patient. Moreover, the presence of int-2 amplification in preinvasive, preinvasive and adjacent invasive lesions, and invasive carcinomas suggest that int-2 could be a marker of genetic instability occurring in early and late stages of tumor development.


Assuntos
Neoplasias da Mama/genética , Carcinoma Intraductal não Infiltrante/genética , Carcinoma Lobular/genética , Carcinoma Medular/genética , Fatores de Crescimento de Fibroblastos/genética , Proteínas Proto-Oncogênicas/genética , Adulto , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Carcinoma Intraductal não Infiltrante/metabolismo , Carcinoma Intraductal não Infiltrante/patologia , Carcinoma Lobular/metabolismo , Carcinoma Lobular/patologia , Carcinoma Medular/metabolismo , Carcinoma Medular/patologia , Feminino , Fator 3 de Crescimento de Fibroblastos , Humanos , Antígeno Ki-67/metabolismo , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase/métodos , Antígeno Nuclear de Célula em Proliferação/metabolismo , Receptores de Estrogênio/genética
19.
Genomics ; 60(1): 105-9, 1999 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-10458916

RESUMO

We report the cDNA cloning and characterization of ZNF236, a novel Kruppel-like zinc-finger gene initially identified by its glucose-regulated expression in human mesangial cells using mRNA differential display. Using the differential display fragment as a probe, we screened a human fetal kidney cDNA library and isolated several clones representing two differently spliced mRNA transcripts, designated ZNF236a and -b. Both transcripts were identical apart from the presence of an additional exon in ZNF236a that truncates the open reading frame. RT-PCR analysis confirmed the expression of both transcripts to be upregulated in human mesangial cells in response to elevated levels of d-glucose. ZNF236a and -b cDNAs encode polypeptides of 174 and 204 kDa, containing 25 and 30 C(2)H(2) zinc-finger motifs, respectively. Northern blot analysis showed that ZNF236 is ubiquitously expressed in all human tissues tested. Expression levels were highest in skeletal muscle and brain, intermediate in heart, pancreas, and placenta, and lowest in kidney, liver, and lung. Southern zoo blot analysis indicated that ZNF236 is conserved in the genomes of all mammalian species tested, but not in yeast. The mapping of ZNF236 to human chromosome 18q22-q23, close to the IDDM6 locus, coupled with the glucose-regulated expression of the gene in human mesangial cells, suggests that ZNF236 may be a candidate gene for diabetic nephropathy.


Assuntos
Proteínas de Transporte/genética , Cromossomos Humanos Par 18/genética , Proteínas de Ligação a DNA/genética , Proteínas Repressoras , Fatores de Transcrição/genética , Dedos de Zinco/genética , Adulto , Processamento Alternativo , Sequência de Aminoácidos , Animais , Northern Blotting , Mapeamento Cromossômico , Clonagem Molecular , DNA Complementar/química , DNA Complementar/genética , DNA Complementar/isolamento & purificação , Feminino , Glucose/fisiologia , Humanos , Hibridização in Situ Fluorescente , Fatores de Transcrição Kruppel-Like , Dados de Sequência Molecular , Isoformas de Proteínas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Distribuição Tecidual
20.
Biochem J ; 316 ( Pt 3): 985-92, 1996 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-8670179

RESUMO

Post-mitotic cultures of human mesangial cells were maintained in media containing 4-30 mM D-glucose for up to 28 days. Changes in mRNA and protein levels for specific macromolecules occurred between 7 and 14 days after initiating hyperglycaemic conditions. Slot blot analysis showed 2-3-fold increases in mRNAs for collagen type I, fibronectin, versican and perlecan, whereas mRNA for decorin was increased by up to 20-fold. Levels of mRNAs for biglycan and syndecan were unaffected by hyperglycaemic culture. Reverse transcriptase PCR (RT-PCR) confirmed that decorin mRNA levels are greatly elevated and also showed increased transcription of the TGF-beta 1 gene in hyperglycaemic cultures. Western analysis and ELISA indicated accumulations of collagen types I and III, laminin and fibronectin in the cell layers and media of hyperglycaemic cultures with increasing time. Type IV collagen did not accumulate in either compartment of hyperglycaemic mesangial cell cultures. Collagen types I, III, and fibronectin did not accumulate in the cell layers of hyperglycaemic human dermal fibroblasts, indicating a cell-specific response in mesangial cultures. Decorin and versican, but not biglycan, were increased in the hyperglycaemic mesangial cell culture media. There were no apparent changes in core proteins for decorin and biglycan in fibroblast media. Transforming growth factor beta 1 (TGF-beta 1) in hyperglycaemic mesangial cell cultures increased 5-fold after 7 days, but decreased thereafter to only approx. 2-fold after 28 days. The changes in TGF-beta 1 mRNA, as detected by RT-PCR, and protein followed one another closely.


Assuntos
Proteínas da Matriz Extracelular/biossíntese , Mesângio Glomerular/metabolismo , Glucose/farmacologia , Hiperglicemia , Córtex Renal/metabolismo , Transcrição Gênica , Sequência de Bases , Células Cultivadas , Primers do DNA , Feminino , Fibroblastos/citologia , Fibroblastos/metabolismo , Mesângio Glomerular/citologia , Humanos , Córtex Renal/citologia , Cinética , Pessoa de Meia-Idade , Mitose , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , RNA Mensageiro/biossíntese , Pele/citologia , Pele/metabolismo , Fatores de Tempo , Transcrição Gênica/efeitos dos fármacos , Fator de Crescimento Transformador beta/biossíntese
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