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The effectiveness of alternate light source (ALS) to fluoresce bone and other materials is well-attested to in a laboratory setting but rarely, if ever, has it been used in field excavation. This study examined the recovery rates of fragmentary bone, fabric, and metal, both with and without the use of an ALS, through practical and controlled excavation experiments with multiple users. All archaeology, including forensic archaeology and crime scene investigation more generally, should account for trace evidence. Currently, there is limited empirical data for the recovery of evidence from excavation, and those studies that do exist, highlight the short-comings in current methods. Six comparable test pits were created, representing empty graves in which only trace evidence remained. Each contained 20 fragments of bone (≤10mm), 16 hair fibres, two pieces of fabric and two lead pieces, which were back-filled and left for over 15 weeks. Three excavators were each tasked with excavating two test pits: one using ALS, one in daylight conditions. The results of the experiment identified some critical aspects of using blue 455nm wavelength ALS in the field, and the importance of experienced practitioners. Sample evidence was small in size and recovery rates were low. In daylight conditions, an average of 46% of trace evidence was identified, while just 40% was recovered using ALS. This excludes hair fibres which were almost undetectable in all conditions. When using ALS, smaller bone fragments were more than twice as likely to be recovered, but less non-fluorescent materials were found. The experience of each excavator had a positive correlation with excavation results. Excavation error rates were calculated, demonstrating that excavation is comparable using either technique, but daylight conditions lead to greater accuracy. The findings suggest that ALS can be used to increase recovery of some evidence types. Test pits provided none of the usual primary evidence associated with graves and excavators had no prior experience of ALS. While retrieval rates were low, almost all recovered items were found in situ and an accurate records maintained. Error rates in forensic archaeology are essential and it is hoped that the method outlined here can be developed towards the establishment of acceptable error rates. While ALS use in forensic archaeology should not be considered a panacea to issues of trace evidence recovery, a combination of well-tested archaeological excavation methods, alongside the implementation of such proven forensic techniques, would likely lead to improved recovery of evidence.
Assuntos
Arqueologia , Fluorescência , Luz , Osso e Ossos , Sepultamento , Ciências Forenses/métodos , Cabelo , Humanos , Metais , TêxteisRESUMO
BACKGROUND: Pain is one of the primary motivations for patients to seek medical advice. Pain location is one element in the process of formulating a diagnosis. AIMS: The purpose of the study is to determine if there is a correlation between the location of pain and the location of pathology in the knees of patients with a suspected meniscus tear. METHODS: From a possible 856 patients referred for arthroscopy, 213 patients consented to be included in the study and 193 (90 %) completed the study. The participating subjects located area of their symptoms on a diagram showing the four aspects of the knee joint. For analysis purposes symptoms were grouped into medial, lateral, posterior, or a combination of these areas. Pathology identified at arthroscopy was recorded on the International Knee Documentation Committee (IKDC) surgical form. The location of knee pathology was divided into medial compartment, lateral compartment or combinations of pathologies. Locations of pain were analysed for an association with the location of pathology found at arthroscopy. RESULTS: Of the 193 subjects who completed the study, 69 (35.7 %) of the subjects presented with one location of pain i.e. medial, lateral or posterior pain and the remaining 124 (64.3 %) had multiple areas. In correlating locations of reported pain with pathology, there was no significant correlation found (p = 0.98). CONCLUSIONS: This study found no direct correlation between the location of pain and the location of pathology in the knee in patients with a suspected meniscus tear.
Assuntos
Artralgia/diagnóstico , Artroscopia , Traumatismos do Joelho/diagnóstico , Articulação do Joelho/patologia , Meniscos Tibiais/patologia , Medição da Dor , Adulto , Idoso , Idoso de 80 Anos ou mais , Artralgia/etiologia , Artralgia/patologia , Artralgia/cirurgia , Feminino , Humanos , Traumatismos do Joelho/complicações , Traumatismos do Joelho/patologia , Traumatismos do Joelho/cirurgia , Articulação do Joelho/cirurgia , Masculino , Meniscos Tibiais/cirurgia , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Ruptura , Lesões do Menisco Tibial , Adulto JovemRESUMO
UNLABELLED: A 19-year-old woman was diagnosed with osteogenesis imperfecta (OI). She had sustained numerous low-trauma fractures throughout her childhood, including a recent pelvic fracture (superior and inferior ramus) following a low-impact fall. She had the classical blue sclerae, and dual energy X-ray absorptiometry (DEXA) bone scanning confirmed low bone mass for her age in the lumbar spine (Z-score was -2.6). However, despite these classical clinical features, the diagnosis of OI had not been entertained throughout the whole of her childhood. Sequencing of her genomic DNA revealed that she was heterozygous for the c.3880_3883dup mutation in exon 50 of the COL1A1 gene. This mutation is predicted to result in a frameshift at p.Thr1295, and truncating stop codon 3 amino acids downstream. To our knowledge, this mutation has not previously been reported in OI. LEARNING POINTS: OI is a rare but important genetic metabolic bone and connective tissue disorder that manifests a diverse clinical phenotype that includes recurrent low-impact fractures.Most mutations that underlie OI occur within exon 50 of the COL1A1 gene (coding for protein constituents of type 1 pro-collagen).The diagnosis of OI is easily missed in its mild form. Early diagnosis is important, and there is a need for improved awareness of OI among health care professionals.OI is a diagnosis of exclusion, although the key diagnostic criterion is through genetic testing for mutations within the COL1A1 gene.Effective management of OI should be instituted through a multidisciplinary team approach that includes a bone specialist (usually an endocrinologist or rheumatologist), a geneticist, an audiometrist and a genetic counsellor. Physiotherapy and orthopaedic surgery may also be required.
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CONTEXT: Dipeptidyl peptidase IV (DPP-IV) inactivates the incretin hormone glucagon-like peptide. It can also affect the orexigenic hormone neuropeptide Y (NPY(1-36)) which is truncated by DPP-IV to NPY(3-36), as a consequence NPY's affinity changes from receptor Y1, which mediates the antilipolytic function of NPY, to other NPY receptors. Little is known whether DPP-IV inhibitors for the treatment of type 2 diabetic (T2DM) patients could influence these pathways. AIMS: To investigate the in vitro effects of NPY with DPP-IV inhibition in isolated abdominal subcutaneous (AbdSc) adipocytes on fat metabolism, and assessment of NPY receptor and DPP-IV expression in adipose tissue (AT). METHODS: Ex vivo human AT was taken from women undergoing elective surgery (body mass index: 27.5 (mean +/- s.d.) +/- 5 kg/m2, age: 43.7 +/- 10 years, n = 36). Isolated AbdSc adipocytes were treated with human recombinant (rh)NPY (1-100 nM) with and without DPP-IV inhibitor (1 M); glycerol release and tissue distribution of DPP-IV, Y1 and Y5 messenger RNA (mRNA) were measured and compared between lean and obese subjects. RESULTS AND CONCLUSION: rhNPY reduced glycerol release, an effect that was further enhanced by co-incubation with a DPP-IV inhibitor [control: 224 (mean +/- s.e.) +/- 37 micromol/l; NPY, 100 nM: 161 +/- 27 micromol/l**; NPY 100 nM/DPP-IV inhibitor, 1 M: 127 +/- 14 micromol/l**; **p < 0.01, n = 14]. DPP-IV was expressed in AbdSc AT and omental AT with relative DPP-IV mRNA expression lower in AbdSc AT taken from obese [77 +/- 6 signal units (SU)] vs. lean subjects (186 +/- 29 SU*, n = 10). Y1 was predominantly expressed in fat and present in all fat depots but higher in obese subjects, particularly the AbdSc AT-depot (obese: 1944 +/- 111 SU vs. lean: 711 +/- 112 SU**, n = 10). NPY appears to be regulated by AT-derived DPP-IV. DPP-IV inhibitors augment the antilipolytic effect of NPY in AT. Further studies are required to show whether this explains the lack of weight loss in T2DM patients treated with DPP-IV inhibitors.
Assuntos
Adipócitos/efeitos dos fármacos , Inibidores da Dipeptidil Peptidase IV/farmacologia , Neuropeptídeo Y/farmacologia , Gordura Subcutânea Abdominal/efeitos dos fármacos , Adipócitos/metabolismo , Adulto , Células Cultivadas , Dipeptidil Peptidase 4/metabolismo , Dipeptidil Peptidase 4/fisiologia , Feminino , Glicerol/metabolismo , Humanos , Lipólise/efeitos dos fármacos , Pessoa de Meia-Idade , Obesidade/metabolismo , Obesidade/patologia , Proteínas Recombinantes/farmacologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Gordura Subcutânea Abdominal/metabolismo , Gordura Subcutânea Abdominal/patologiaRESUMO
OBJECTIVES: Ghrelin, an important central acting orexigenic hormone, is predominantly secreted in the gastrointestinal tract. However little is known about the action of ghrelin in human adipose tissue (AT). AIM: To study the expression of ghrelin in AT, the effects of octanoyl-(OTG) and des-acyl (DSG) ghrelin on lipolysis and lipogenesis, leptin release and potential peripheral signalling through the Y1 receptor. METHODS: Ex vivo human AT was obtained from women undergoing elective surgery (46 (mean +/- SD) 6.8 years, body mass index (BMI): 25.6 +/- 5.0 kg/m(2), n = 20). Abdominal-subcutaneous (AbdSc) adipocytes were isolated and treated with recombinant human (rh) OTG and DSG to assess lipid metabolism leptin release and the influence of Y1-receptor blocker. RESULTS: Ghrelin was expressed in AbdScAT and negatively correlated with BMI (lean: 3.6 +/- 0.74 optical-density-units (OD), obese: 1.64 +/- 0.45 OD, *P < 0.05). Only DSG significantly suppressed glycerol release (Control (C): 286 +/- 58 microl/l; DSG 1 nm: 224 +/- 38 microl/l downward arrow*; DSG 100 nm: 172 +/- 13 microl/l downward arrow*,* downward arrow P < 0.05, n = 7) and reduced hormone sensitive lipase expression (C: 1.0 +/- 0.3 OD; DSG 1 nm: 0.8 +/- 0.3 OD downward arrow*; DSG 100 nm: 0.6 +/- 0.1 OD downward arrow*, n = 4). However, both isoforms increased lipoprotein lipase expression (C: 1.0 +/- 0.3OD; DSG 100 nm: 0.2 +/- 0.4 OD upward arrow*; OTG 100 nm: 2.5 +/- 0.3 OD upward arrow*, n = 4), whilst blockade of Y1 eliminated this effect in both. Leptin was down-regulated by DSG only (DSG 1 nm: 5.3 +/- 0.7 ng/ml; DSG 100 nm: 4.1 +/- 0.7 ng/ml*) and was significant after BMI adjustment (P = 0.029). CONCLUSION: Ghrelin was expressed in human AbdSc AT. In vitro, both OGT and DSG appear to mediate fat deposition with the lipogenic effects in part mediated by the Y1 receptor, whilst the influence of DSG affected lipolysis, lipogenesis and leptin secretion. Taken together, these studies support a local action for ghrelin isoforms on lipid and adipokine metabolism that further supports a cross talk between organs.
Assuntos
Grelina/metabolismo , Gordura Subcutânea/metabolismo , Adipócitos/citologia , Adipócitos/efeitos dos fármacos , Adipócitos/metabolismo , Adulto , Arginina/análogos & derivados , Arginina/farmacologia , Índice de Massa Corporal , Células Cultivadas , Feminino , Grelina/farmacologia , Humanos , Leptina/metabolismo , Lipogênese/efeitos dos fármacos , Lipogênese/fisiologia , Lipólise/efeitos dos fármacos , Lipólise/fisiologia , Lipase Lipoproteica/metabolismo , Pessoa de Meia-Idade , Receptores de Neuropeptídeo Y/antagonistas & inibidores , Gordura Subcutânea/citologia , Gordura Subcutânea/efeitos dos fármacosRESUMO
CONTEXT: Visceral adipose tissue (AT) is known to confer a significantly higher risk of type 2 diabetes and cardiovascular disease. Epicardial AT has been shown to be related to cardiovascular disease and myocardial function through unidentified mechanisms. Epicardial AT expresses an inflammatory profile of proteins; however, the mechanisms responsible are yet to be elucidated. OBJECTIVES: The objectives of the study were to: 1) examine key mediators of the nuclear factor-kappaB (NFkappaB) and c-Jun N-terminal kinase (JNK) pathways in paired epicardial and gluteofemoral (thigh) AT from coronary artery disease (CAD) and control patients and 2) investigate circulating endotoxin levels in CAD and control subjects. DESIGN: Serums and AT biopsies (epicardial and thigh) were obtained from CAD (n = 16) and non-CAD (n = 18) patients. Inflammation was assessed in tissue and serum samples through Western blot, real-time PCR, ELISAs, and activity studies. RESULTS: Western blotting showed epicardial AT had significantly higher NFkappaB, inhibitory-kappaB kinase (IKK)-gamma, IKKbeta, and JNK-1 and -2 compared with thigh AT. Epicardial mRNA data showed strong correlations between CD-68 and toll-like receptor-2, toll-like receptor-4, and TNF-alpha. Circulating endotoxin was elevated in patients with CAD compared with matched controls [CAD: 6.80 +/- 0.28 endotoxin unit(EU)/ml vs. controls: 5.52 +/- 0.57 EU/ml; P<0.05]. CONCLUSION: Epicardial AT from patients with CAD shows increased NFkappaB, IKKbeta, and JNK expression compared with both CAD thigh AT and non-CAD epicardial AT, suggesting a depot-specific as well as a disease-linked response to inflammation. These studies implicate both NFkappaB and JNK pathways in the inflammatory profile of epicardial AT and highlight the role of the macrophage in the inflammation within this tissue.
Assuntos
Tecido Adiposo/fisiologia , Doença da Artéria Coronariana/complicações , Inflamação/etiologia , Proteínas Quinases JNK Ativadas por Mitógeno/fisiologia , NF-kappa B/fisiologia , Pericárdio/metabolismo , Idoso , Antígenos CD/genética , Antígenos de Diferenciação Mielomonocítica/genética , Endotoxinas/sangue , Feminino , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/análise , Masculino , Pessoa de Meia-Idade , NF-kappa B/análise , Fosforilação , RNA Mensageiro/análise , Receptor 4 Toll-Like/genética , Fator de Necrose Tumoral alfa/genéticaRESUMO
OBJECTIVES: To determine, first, the effects of menopausal status on circulating calcitonin gene-related peptide (CGRP) levels and, second, the correlation between circulating CGRP levels and biomarkers for cardiovascular disease. METHODS: Cross-sectional study of healthy premenopausal and postmenopausal women volunteers and women admitted for elective benign abdominal surgery in a district general hospital. All women were non-smokers, had no history of endocrinological problems and were not receiving any hormone therapy. Fasting blood samples (premenopausal (n = 45): follicle stimulating hormone (FSH) < 20 IU/l, estradiol (mean +/- SEM) 440.33 +/- 51.82 pmol/l; postmenopausal women (n = 28): FSH > 20 IU/l, estradiol 93.79 +/- 17.40 pmol/l) were analyzed for CGRP, resistin, leptin, adiponectin, insulin and lipids using ELISA and immunoassays. RESULTS: Mean circulating CGRP levels were higher in the postmenopausal women compared with premenopausal women (pre: 41.79 +/- 9.01 pg/ml, post: 138.14 +/- 45.75 pg/ml; p = 0.047). Among women who were experiencing hot flushes, the postmenopausal women had significantly higher CGRP levels than the premenopausal women (pre: 21.98 +/- 4.95 pg/ml, post: 171.08 +/- 61.80 pg/ml; p = 0.028). Serum CGRP levels positively correlated with serum insulin levels (r = 0.652, p = 0.016) and HOMA index (r = 0.54, p < 0.001). CONCLUSION: These data show that circulating CGRP levels are influenced by menopausal status and suggest additional mechanisms through which increased risk of hyperinsulinemia and cardiovascular disease may arise in postmenopausal women.
Assuntos
Adipocinas/sangue , Peptídeo Relacionado com Gene de Calcitonina/sangue , Pós-Menopausa/sangue , Pré-Menopausa/sangue , Adulto , Índice de Massa Corporal , Colesterol/sangue , Estudos Transversais , Estradiol/sangue , Feminino , Hormônio Foliculoestimulante/sangue , Fogachos/sangue , Humanos , Insulina/sangue , Resistência à Insulina , Pessoa de Meia-Idade , Triglicerídeos/sangueRESUMO
BACKGROUND: Orlistat has been shown to increase adiponectin and reduce progression to type 2 diabetes in obese Caucasians. Some effects of orlistat are thought to be independent of weight loss by altering gut flora and the production of endotoxin lipopolysaccharide (LPS). We studied the effect of dietary treatment with and without orlistat in South Asian individuals with impaired glucose tolerance (IGT) on adiponectin and inflammatory markers including LPS. METHODS: South Asian individuals were randomised to either dietary treatment with orlistat or dietary treatment alone. At the end of 12 months, a comparison was made between the two groups for differences in anthropomorphic measurements and serum markers. RESULTS: Three hundred and five individuals underwent oral glucose tolerance test of whom 40 had IGT. Complete baseline and 1-year data was available for 31 patients. After 1 year, patients in the orlistat group demonstrated a greater but insignificant decrease in weight (4.5 +/- 0.1 kg), and a significant increase in adiponectin (6.73 +/- 3.2 microg/ml) and decrease in LPS (4.55 +/- 1.98 EU/ml) compared with- the diet-alone group. In the orlistat group the reduction in LPS was correlated with the increase in adiponectin (p < 0.005). CONCLUSION: The increase in adiponectin levels in the orlistat group would suggest that orlistat may reduce the progression to type 2 diabetes in South Asian individuals by raising serum adiponectin. The finding that LPS levels are also reduced by orlistat and that this reduction correlates with the increase in adiponectin raises the possibility that the increase in adiponectin may be mediated via an effect on LPS levels.
Assuntos
Adipocinas/sangue , Intolerância à Glucose/tratamento farmacológico , Lactonas/uso terapêutico , Lipopolissacarídeos/sangue , Adulto , Terapia Combinada , Feminino , Seguimentos , Intolerância à Glucose/sangue , Intolerância à Glucose/dietoterapia , Humanos , Lipídeos/sangue , Masculino , OrlistateRESUMO
tRNA identity elements assure the correct aminoacylation of tRNAs by the aminoacyl-tRNA synthetases with the cognate amino acid. The tRNA(Gly)/glycyl-tRNA sythetase system is member of the so-called 'class II system' in which the tRNA determinants consist of rather simple elements. These are mostly located in the tRNA acceptor stem and in the glycine case additionally the discriminator base at position 73 is required. Within the glycine-tRNA synthetases, the archaebacterial/human and the eubacterial sytems differ with respect to their protein structures and the required tRNA identity elements, suggesting a unique evolutionary divergence. In this study, we present a comparison between the crystal structures of the eubacterial Escherichia coli and the human tRNA(Gly) acceptor stem microhelices and their surrounding hydration patterns.
Assuntos
Escherichia coli/genética , Glicina-tRNA Ligase/química , RNA de Transferência de Glicina/química , Humanos , Modelos Moleculares , Conformação de Ácido Nucleico , Água/químicaRESUMO
UNLABELLED: Type 2 diabetes (T2DM) is associated with chronic low-grade inflammation. Adipose tissue (AT) may represent an important site of inflammation. 3T3-L1 studies have demonstrated that lipopolysaccharide (LPS) activates toll-like receptors (TLRs) to cause inflammation. For this study, we 1) examined activation of TLRs and adipocytokines by LPS in human abdominal subcutaneous (AbdSc) adipocytes, 2) examined blockade of NF-kappaB in human AbdSc adipocytes, 3) examined the innate immune pathway in AbdSc AT from lean, obese, and T2DM subjects, and 4) examined the association of circulating LPS in T2DM subjects. The findings showed that LPS increased TLR-2 protein expression twofold (P<0.05). Treatment of AbdSc adipocytes with LPS caused a significant increase in TNF-alpha and IL-6 secretion (IL-6, CONTROL: 2.7+/-0.5 vs. LPS: 4.8+/-0.3 ng/ml; P<0.001; TNF-alpha, CONTROL: 1.0+/-0.83 vs. LPS: 32.8+/-6.23 pg/ml; P<0.001). NF-kappaB inhibitor reduced IL-6 in AbdSc adipocytes ( CONTROL: 2.7+/-0.5 vs. NF-kappaB inhibitor: 2.1+/-0.4 ng/ml; P<0.001). AbdSc AT protein expression for TLR-2, MyD88, TRAF6, and NF-kappaB was increased in T2DM patients (P<0.05), and TLR-2, TRAF-6, and NF-kappaB were increased in LPS-treated adipocytes (P<0.05). Circulating LPS was 76% higher in T2DM subjects compared with matched controls. LPS correlated with insulin in controls (r=0.678, P<0.0001). Rosiglitazone (RSG) significantly reduced both fasting serum insulin levels (reduced by 51%, P=0.0395) and serum LPS (reduced by 35%, P=0.0139) in a subgroup of previously untreated T2DM patients. In summary, our results suggest that T2DM is associated with increased endotoxemia, with AT able to initiate an innate immune response. Thus, increased adiposity may increase proinflammatory cytokines and therefore contribute to the pathogenic risk of T2DM.
Assuntos
Diabetes Mellitus Tipo 2/imunologia , Imunidade Inata/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Obesidade/imunologia , Gordura Subcutânea Abdominal/efeitos dos fármacos , Adipócitos Brancos/efeitos dos fármacos , Adipócitos Brancos/imunologia , Adipócitos Brancos/metabolismo , Adulto , Idoso , Células Cultivadas , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fator 88 de Diferenciação Mieloide/metabolismo , NF-kappa B/antagonistas & inibidores , Obesidade/sangue , Obesidade/patologia , Gordura Subcutânea Abdominal/imunologia , Gordura Subcutânea Abdominal/metabolismo , Fator 6 Associado a Receptor de TNF/metabolismo , Receptores Toll-Like/metabolismoRESUMO
The renin-angiotensin system is an important regulator of blood pressure, and blockade of this system improves blood pressure in obesity and type 2 diabetes. Recently, components of the system have been described in adipose tissue. However, to date no study has investigated the influence of varying insulin concentrations on angiotensinogen (AGT) protein expression in human subcutaneous abdominal fat. Isolated subcutaneous adipocytes were treated with insulin (1-1000 nm) for 48 h. As part of the studies, a novel AGT antibody was developed and validated by Western blotting and immunohistochemistry. Western blotting was performed on the protein extracted from the adipocytes treated with insulin to determine AGT expression. Increasing doses of insulin raised AGT protein expression in a dose-dependent manner (control 1.0 +/- 0.0 (mean +/- s.e.) - protein expression standardized relative to control; 1 nm insulin: 2.64 +/- 0.0.32 upward arrow ***; 100 nm insulin: 4.37 +/- 0.57 upward arrow ***; 1000 nm insulin: 6.50 +/- 0.97 upward arrow ***; ***p < 0.001, n = 3). In conclusion, increasing insulin doses stimulates AGT production. In this study, protein analysis suggests that hyperinsulinaemia may be an important factor in obesity-related hypertension.
Assuntos
Adipócitos/efeitos dos fármacos , Angiotensinogênio/metabolismo , Insulina/farmacologia , Abdome , Adipócitos/metabolismo , Tecido Adiposo/metabolismo , Adulto , Western Blotting , Células Cultivadas , Relação Dose-Resposta a Droga , Feminino , Humanos , Rim/metabolismo , Fígado/metabolismo , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
OBJECTIVE: The aim of this study was to investigate the effect of insulin and an insulin-sensitizing agent, rosiglitazone (RSG), on the production of plasminogen-activator inhibitor-1 (PAI-1) in isolated subcutaneous abdominal adipocytes. Human tissue-type plasminogen activator (t-PA) was also measured to assess changes in overall thrombotic risk. METHODS: The mean depot-specific expression of PAI-1 and t-PA mRNA (n = 42) in subcutaneous abdominal (n = 21), omental (n = 10) and thigh (n = 11) adipose tissue depots was examined. Furthermore, subcutaneous adipocytes were treated with insulin, RSG and insulin in combination with RSG (10-8 m) for 48 h. Conditioned media were collected and enzyme-linked immunosorbent assays performed for PAI-1 and t-PA (n = 12) antigen. PAI-1 and t-PA mRNA levels were also assessed. RESULTS: PAI-1 mRNA levels were significantly higher in subcutaneous and omental abdominal tissue than in thigh fat (p = 0.037 and p = 0.014). No change in t-PA mRNA expression between the adipose tissue depots was observed. Insulin stimulated PAI-1 protein secretion in a concentration-dependent manner in adipocytes (control: 68.3 +/- 1.2 ng/ml (s.e.m.); 10 nm insulin: 73.7 +/- 3.8 ng/ml upward arrow; 100 nm insulin: 86.8 +/- 4.1 ng/ml upward arrow **; 1000 nm insulin: 102.0 +/- 4.8 ng/ml upward arrow ***; **p < 0.01, ***p < 0.001). In contrast, insulin + RSG (10-8 m) reduced PAI-1 production relative to insulin alone (***p < 0.001), whilst RSG alone reduced PAI-1 protein secretion in a concentration-dependent manner (RSG at 10-10 m: 50.4 +/- 2.87 ng/ml downward arrow ***; RSG at 10-5 m: 30.3 +/- 2.0 ng/ml downward arrow ***; p < 0.001). No difference was observed between control and treatments for t-PA secretion (range 7-11 ng/ml). CONCLUSIONS: Insulin stimulated PAI-1 secretion, whilst RSG reduced both PAI-1 secretion alone and in combination with insulin. These data suggest that adipose tissue may contribute significantly to the elevated circulating PAI-1 in obesity. Therefore, RSG's effects on PAI-1 production in adipose tissue may contribute to the fall in circulating PAI-1 levels observed in patients receiving RSG therapy.
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Adipócitos/efeitos dos fármacos , Hipoglicemiantes/farmacologia , Inibidor 1 de Ativador de Plasminogênio/biossíntese , Tiazolidinedionas/farmacologia , Adipócitos/metabolismo , Tecido Adiposo/citologia , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/metabolismo , Adulto , Células Cultivadas , Técnicas de Cultura , Relação Dose-Resposta a Droga , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Insulina/farmacologia , Masculino , Pessoa de Meia-Idade , Inibidor 1 de Ativador de Plasminogênio/genética , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rosiglitazona , Ativador de Plasminogênio Tecidual/biossíntese , Ativador de Plasminogênio Tecidual/genéticaRESUMO
Clinical observations suggest a role for testosterone in the accumulation of central adiposity and with an associated increased risk of disease. To date, no human study has analysed the role of dihydrotestosterone (DHT) on adipose tissue mass regulation in vitro. This study investigated the role of DHT and androgen receptors (AR) in the regulation of lipolysis and lipogenesis by examining the key enzymes hormone sensitive lipase (HSL) and lipoprotein lipase (LPL) respectively. Isolated abdominal subcutaneous adipocytes (Scad) (n = 15) were treated with either DHT (10(-7)-10(-9) m), an antiandrogen, flutamide (FLT: 10(-7)-10(-9) m) or a combination of DHT (10(-7)-10(-9) m) with FLT (10(-8) m). Relative protein expression of HSL, LPL and AR was determined. In Scad, DHT inhibited HSL expression maximally at 10(-9) m (0.7 +/- 0.4**; p < 0.01**) compared with control (control: 1.0 +/- (s.e.m.) 0.0), whereas LPL protein expression was stimulated at DHT10(-9) m (2.22 +/- 0.48*; p < 0.05*). Glycerol release assay results correlated with HSL expression data. LPL expression was reduced at all doses with combinations of DHT + FLT compared with DHT alone. Androgen receptor expression studies showed an inverse correlation with DHT, whereas DHT + FLT reduced AR expression. These studies indicate that DHT may alter HSL and LPL expression, whereas only LPL expression appears mediated by AR. These findings suggest a physiological role for DHT in the control of adipose tissue mass in women, and indicate that androgens may also play an important role in regulating lipid metabolism.
Assuntos
Adipócitos/enzimologia , Tecido Adiposo/enzimologia , Di-Hidrotestosterona/farmacologia , Flutamida/farmacologia , Lipase Lipoproteica/metabolismo , Receptores Androgênicos/metabolismo , Esterol Esterase/metabolismo , Células Cultivadas , Feminino , Humanos , Pessoa de Meia-Idade , Receptores Androgênicos/efeitos dos fármacosRESUMO
Resistin, an adipocyte-derived cytokine, causes insulin resistance and glucose intolerance in mice. We investigated whether resistin expression was higher in human abdominal adipose tissue than other adipose tissue depots. We extracted RNA from 32 adipose tissue samples (13 subcutaneous abdominal, seven omentum, six thigh, and six breast). Quantitative PCR was used to determine resistin mRNA expression. Resistin mRNA concentrations were similar in both the subcutaneous abdominal and omental depots. The abdominal depots showed a 418% increase in resistin mRNA expression compared with the thigh. Increased resistin expression in abdominal fat could explain the increased risk of type 2 diabetes associated with central obesity.
Assuntos
Tecido Adiposo/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Hormônios Ectópicos/genética , Resistência à Insulina/genética , Peptídeos e Proteínas de Sinalização Intercelular , Obesidade/metabolismo , RNA Mensageiro/genética , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , RNA Mensageiro/isolamento & purificação , ResistinaRESUMO
Adiponectin is an adipocyte-derived hormone associated with insulin sensitivity and atherosclerotic risk. As central rather than gluteofemoral fat is known to increase the risk of type 2 diabetes and cardiovascular disease, we investigated the mRNA and protein expression of adiponectin in human adipose tissue depots. RNA was extracted from 46 human adipose tissue samples from non-diabetic subjects aged 44.33 +/- 12.4 with a BMI of 28.3 +/- 6.0 (mean +/- SD). The samples were as follows: 21 abdominal subcutaneous, 13 omentum, 6 thigh; samples were also taken from diabetic subjects aged 66.6 +/- 7.5 with BMI 28.9 +/- 3.17; samples were: 6 abdominal subcutaneous; 3 thigh. Quantitative PCR and Western analysis was used to determine adiponectin content. Protein content studies determined that when compared with non-diabetic abdominal subcutaneous adipose tissue (Abd Sc AT) (values expressed as percentage relative to Abd Sc AT -100 %). Adiponectin protein content was significantly lower in non-diabetic omental AT (25 +/- 1.6 %; p < 0.0001, n = 6) and in Abd Sc AT from diabetic subjects (36 +/- 1.5 %; p < 0.0001, n = 4). In contrast, gluteal fat maintained high adiponectin protein content from non-diabetic patients compared with diabetic patients. An increase in BMI was associated with lower adiponectin protein content in obese ND Abd Sc AT (25 +/- 0.4 %; p < 0.0001). These findings were in agreement with the mRNA expression data. In summary, this study indicates that adiponectin protein content in non-diabetic subjects remains high in abdominal subcutaneous fat, including gluteal fat, explaining the high serum adiponectin levels in these subjects. Omental fat, however, expresses little adiponectin. Furthermore, abdominal and gluteal subcutaneous fat appears to express significantly less adiponectin once diabetic status is reached. In conclusion, the adipose tissue depot-specific expression of adiponectin may influence the pattern of serum adiponectin concentrations and subsequent disease risk.
Assuntos
Tecido Adiposo/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular , Obesidade , Proteínas/metabolismo , Adipócitos/metabolismo , Adiponectina , Humanos , Imuno-Histoquímica , Técnicas In Vitro , Omento/citologia , Omento/metabolismo , RNA Mensageiro/análise , Tela Subcutânea/metabolismo , Coxa da Perna , Distribuição TecidualRESUMO
Body image perception was measured in 50 women with bulima nervosa and 19 age and weight matched female controls, using a visual size estimation apparatus. Both groups overestimated body widths, but not the width of a neutral object, and whilst there was a trend for bulimics to overestimate more than controls this did not reach significance. The part of the body most overestimated corresponded to the part most disliked in only a third of both groups. The bulimics without a previous history of anorexia nervosa overestimated body width more than those with such a history; this may be related to the fact that the former had a significantly greater weight index. Bulimics who were within 5% of mean-matched population weight overestimated body width less than the others, this difference reaching significance when compared with the heavier groups; a similar, but non-significant, trend was demonstrated in controls. This may be linked to a greater dissatisfaction with body size. Duration of illness, frequency of bingeing and self-induced vomiting were not shown significantly to alter body size estimation. The bulimics who completed a 10-session outpatient treatment programme subsequently demonstrated a significant decrease in overestimation of waist and hip width.
Assuntos
Imagem Corporal , Transtornos da Alimentação e da Ingestão de Alimentos/psicologia , Hiperfagia/psicologia , Adulto , Anorexia Nervosa/psicologia , Estatura , Peso Corporal , Feminino , HumanosRESUMO
A monoclonal antibody, MID 2, which reacts with an epitope common to all human leucocytes, was used to show that the leucocyte common antigen can be resolved into four glycoprotein components with molecular weights of 220 K, 200 K, 180 K and 160 K. The 200, 180 and 160 K peptides were all present to various extents in the T and B lymphoblastoid cell lines examined, but the 220 K component was only detected in the B-cell lines. The 220 K glycoprotein was also lacking in human thymocytes, but evidence of its expression in peripheral blood T lymphocytes was obtained, suggesting that it might be acquired as the cells differentiated. The four glycoproteins isolated from tonsil cells gave similar patterns on peptide mapping by the Cleveland enzymatic method or by cyanogen bromide cleavage, indicating extensive homology. It is conceivable that they differ from each other by the acquisition of similar repeating domain sequences of approximately 20 K. Alternatively, they may share a common peptide structure but differ in their electrophoretic behaviour in SDS gels owing to differences in carbohydrate; if this is the case, however, experiments with tunicamycin suggest that O-linked oligosaccharides must be involved.