RESUMO
BACKGROUND: Early detection of autoimmune Addison's disease (AAD) is important as delay in diagnosis may result in a life-threatening adrenal crisis and death. The classical clinical picture of untreated AAD is well-described, but methodical investigations are scarce. OBJECTIVE: Perform a retrospective audit of patient records with the aim of identifying biochemical markers for early diagnosis of AAD. MATERIAL AND METHODS: A multicentre retrospective study including 272 patients diagnosed with AAD at hospitals in Norway and Sweden during 1978-2016. Scrutiny of medical records provided patient data and laboratory values. RESULTS: Low sodium occurred in 207 of 247 (84%), but only one-third had elevated potassium. Other common nonendocrine tests were largely normal. TSH was elevated in 79 of 153 patients, and hypoglycaemia was found in 10%. Thirty-three per cent were diagnosed subsequent to adrenal crisis, in whom electrolyte disturbances were significantly more pronounced (P < 0.001). Serum cortisol was consistently decreased (median 62 nmol L-1 [1-668]) and significantly lower in individuals with adrenal crisis (38 nmol L-1 [2-442]) than in those without (81 nmol L-1 [1-668], P < 0.001). CONCLUSION: The most consistent biochemical finding of untreated AAD was low sodium independent of the degree of glucocorticoid deficiency. Half of the patients had elevated TSH levels. Only a minority presented with marked hyperkalaemia or other nonhormonal abnormalities. Thus, unexplained low sodium and/or elevated TSH should prompt consideration of an undiagnosed AAD, and on clinical suspicion bring about assay of cortisol and ACTH. Presence of 21-hydroxylase autoantibodies confirms autoimmune aetiology. Anticipating additional abnormalities in routine blood tests may delay diagnosis.
Assuntos
Doença de Addison/diagnóstico , Diagnóstico Precoce , Doença de Addison/sangue , Doença de Addison/complicações , Adolescente , Adulto , Idoso , Biomarcadores/sangue , Glicemia/metabolismo , Criança , Pré-Escolar , Feminino , Humanos , Hidrocortisona/sangue , Hiperpotassemia/etiologia , Hipoglicemia/etiologia , Hiponatremia/etiologia , Masculino , Pessoa de Meia-Idade , Potássio/sangue , Estudos Retrospectivos , Sódio/sangue , Tireotropina/sangue , Adulto JovemRESUMO
BACKGROUND: Autoimmune disease is one of the leading causes of morbidity and mortality worldwide. In Addison's disease, the adrenal glands are targeted by destructive autoimmunity. Despite being the most common cause of primary adrenal failure, little is known about its aetiology. METHODS: To understand the genetic background of Addison's disease, we utilized the extensively characterized patients of the Swedish Addison Registry. We developed an extended exome capture array comprising a selected set of 1853 genes and their potential regulatory elements, for the purpose of sequencing 479 patients with Addison's disease and 1394 controls. RESULTS: We identified BACH2 (rs62408233-A, OR = 2.01 (1.71-2.37), P = 1.66 × 10-15 , MAF 0.46/0.29 in cases/controls) as a novel gene associated with Addison's disease development. We also confirmed the previously known associations with the HLA complex. CONCLUSION: Whilst BACH2 has been previously reported to associate with organ-specific autoimmune diseases co-inherited with Addison's disease, we have identified BACH2 as a major risk locus in Addison's disease, independent of concomitant autoimmune diseases. Our results may enable future research towards preventive disease treatment.
Assuntos
Doença de Addison/genética , Fatores de Transcrição de Zíper de Leucina Básica/genética , Exoma/genética , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Haplótipos , Antígenos de Histocompatibilidade Classe II/genética , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Análise de Sequência , Adulto JovemRESUMO
The adrenalitis found in autoimmune Addison's disease (AAD) is considered having a Th1-driven pathogenesis. Circulating Th1- and Th2-associated chemokines responsible for the trafficking of leukocytes to inflammatory sites are markers for the Th1/Th2 balance. The aim of the study was to assess if the same daily hydrocortisone dose of 30 mg given in either 2 or 4 doses to patients with AAD could affect the Th1/Th2 balance of circulating chemokines.Fifteen patients (6 women) with AAD were included in this randomised, placebo controlled, double blind cross-over study. Samples for chemokines, Th1-associated (CXCL10, CXCL11) and Th2-associated (CCL17, CCL22), were drawn 5 times during a 24-h period at the end of each treatment period and analysed with Luminex. Seven control subjects did the same diurnal blood sampling once. Subjects with AAD had higher median diurnal levels of the Th1-associated chemokines than controls, CXCL10 [43 (33-56) pg/ml vs. 22 (19-34) pg/ml, p<0.01] and CXCL11 [37 (29-48) pg/ml vs. 16 (9-24) pg/ml, p<0.001], whereas no significant difference was found regarding the Th2-related chemokines. Similar chemokine levels were found when the same hydrocortisone dose of 30 mg was divided in 2 or 4 doses. Levels of CXCL11 correlated negatively with scores of SF-36 domains (high score indicate better health) of General Health (GH) and total score for Physical Component Summary (PCS), and these negative correlations were most pronounced at 04:00 h on the 2-dose regimen. Patients with AAD have a dominant Th1 chemokine profile that partially correlates to reduced quality of life.
Assuntos
Doença de Addison/sangue , Doença de Addison/imunologia , Quimiocinas/sangue , Hidrocortisona/uso terapêutico , Qualidade de Vida , Células Th1/imunologia , Células Th2/imunologia , Doença de Addison/tratamento farmacológico , Doença de Addison/fisiopatologia , Adulto , Antropometria , Estudos de Casos e Controles , Ritmo Circadiano/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Humanos , Hidrocortisona/sangue , Hidrocortisona/farmacologia , Masculino , Pessoa de Meia-Idade , Inquéritos e QuestionáriosRESUMO
An impaired body balance has been found in Turner syndrome (TS) in clinical tests like Rombergs's test and walking on a balance beam. The aim of the study was to assess postural balance in TS subjects with specific balance testing using dynamic posturography and relate to body composition. Nineteen TS subjects (20-57 years) were included. Balance was measured with dynamic posturography (Equitest) and compared with 19 sex and age-matched controls (22-59 years). Equitest, visual, vestibular, and somatosensory systems were provoked with increasing difficulty (6 tests, SO1-SO6) and body sway was measured with a dual forceplate. Body composition was measured with DXA. No difference was found between the TS subjects and the controls on fixed platform with open eyes (SO1), with closed eyes (SO2), with stable platform and visual disorientation (SO3), or on unstable platform with open eyes (SO4). In the difficult tests on unstable platform the TS subjects did worse compared with controls both in the test with eyes closed (SO5), p<0.01, and in the test with visual disorientation (SO6), p<0.05. Composite (a merge of all six recordings) was significantly lower in the TS-group, p<0.05. In the TS group high total body weight was related to worse outcome on tests SO5, SO6, and composite, while total bone mass, age, height, or waist showed no significant association with balance scores. Our findings indicate that TS could have an increased risk for falling due to impaired ability to manage complex coordination tasks.
Assuntos
Equilíbrio Postural , Síndrome de Turner/fisiopatologia , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Sotrastaurin, a selective protein-kinase-C inhibitor, blocks early T-cell activation through a calcineurin-independent mechanism. In this study, de novo renal transplant recipients with immediate graft function were randomized 1:2 to tacrolimus (control, n = 44) or sotrastaurin (300 mg b.i.d.; n = 81). All patients received basiliximab, mycophenolic acid (MPA) and steroids. The primary endpoint was the composite of treated biopsy-proven acute rejection (BPAR), graft loss, death or lost to follow-up at month 3. The main safety assessment was estimated glomerular filtration rate (eGFR); modification of diet in renal disease (MDRD) at month 3. Composite efficacy failure at month 3 was higher for the sotrastaurin versus control regimen (25.7% vs. 4.5%, p = 0.001), driven by higher BPAR rates (23.6% vs. 4.5%, p = 0.003), which led to early study termination. Median (± standard deviation [SD]) eGFR was higher for sotrastaurin versus control at all timepoints from day 7 (month 3: 59.0 ± 22.3 vs. 49.5 ± 17.7 mL/min/1.73 m(2) , p = 0.006). The most common adverse events were gastrointestinal disorders (control: 63.6%; sotrastaurin: 88.9%) which led to study-medication discontinuation in two sotrastaurin patients. This study demonstrated a lower degree of efficacy but better renal function with the calcineurin-inhibitor-free regimen of sotrastaurin+MPA versus the tacrolimus-based control. Ongoing studies are evaluating alternative sotrastaurin regimens.
Assuntos
Transplante de Rim/fisiologia , Proteína Quinase C/antagonistas & inibidores , Pirróis/uso terapêutico , Quinazolinas/uso terapêutico , Adulto , Inibidores de Calcineurina , Feminino , Humanos , Masculino , Ácido Micofenólico/uso terapêutico , Tacrolimo/uso terapêuticoRESUMO
The Semliki Forest virus (SFV) directs the synthesis of a heterodimeric membrane protein complex which is used for virus membrane assembly during budding at the surface of the infected cell, as well as for low pH-induced membrane fusion in the endosomes when particles enter new host cells. Existing evidence suggests that the E1 protein subunit carries the fusion potential of the heterodimer, whereas the E2 subunit, or its intracellular precursor p62, is required for binding to the nucleocapsid. We show here that during virus uptake into acidic endosomes the original E2E1 heterodimer is destabilized and the E1 proteins form new oligomers, presumably homooligomers, with altered E1 structure. This altered structure of E1 is specifically recognized by a monoclonal antibody which can also inhibit penetration of SFV into host cells as well as SFV-mediated cell-cell fusion, thus suggesting that the altered E1 structure is important for the membrane fusion. These results give further support for a membrane protein oligomerization-mediated control mechanism for the membrane fusion potential in alphaviruses.
Assuntos
Fusão de Membrana , Vírus da Floresta de Semliki/fisiologia , Proteínas do Envelope Viral/fisiologia , Proteínas Virais de Fusão/fisiologia , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Antivirais/imunologia , Linhagem Celular , Cricetinae , Endocitose , Epitopos , Imunofluorescência , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Substâncias Macromoleculares , Monensin/farmacologia , Conformação Proteica , Vírus da Floresta de Semliki/imunologia , Vírus da Floresta de Semliki/ultraestrutura , Tripsina/farmacologia , Proteínas do Envelope Viral/imunologia , Proteínas do Envelope Viral/ultraestruturaRESUMO
The orientation of signal-anchor proteins in the endoplasmic reticulum membrane is largely determined by the charged residues flanking the apolar, membrane-spanning domain and is influenced by the folding properties of the NH2-terminal sequence. However, these features are not generally sufficient to ensure a unique topology. The topogenic role of the hydrophobic signal domain was studied in vivo by expressing mutants of the asialoglycoprotein receptor subunit H1 in COS-7 cells. By replacing the 19-residue transmembrane segment of wild-type and mutant H1 by stretches of 7-25 leucine residues, we found that the length and hydrophobicity of the apolar sequence significantly affected protein orientation. Translocation of the NH2 terminus was favored by long, hydrophobic sequences and translocation of the COOH terminus by short ones. The topogenic contributions of the transmembrane domain, the flanking charges, and a hydrophilic NH2-terminal portion were additive. In combination these determinants were sufficient to achieve unique membrane insertion in either orientation.
Assuntos
Glicoproteínas de Membrana/metabolismo , Sinais Direcionadores de Proteínas/química , Receptores de Superfície Celular/metabolismo , Sequência de Aminoácidos , Animais , Receptor de Asialoglicoproteína , Transporte Biológico , Células COS , Membrana Celular/metabolismo , Chlorocebus aethiops , Hexosaminidases/farmacologia , Leucina/química , Dados de Sequência Molecular , Peso Molecular , Solubilidade , Relação Estrutura-Atividade , TripsinaRESUMO
The Semliki Forest virus (SFV) spike is composed of three copies of a membrane protein heterodimer. The two subunits of this heterodimer (p62 and E1) are synthesized sequentially from a common mRNA together with the capsid (C) in the order C-p62-E1. In this work heterodimerization of the spike proteins has been studied in BHK 21 cells. The results indicate that: (a) the polyprotein is cotranslationally cleaved into individual chains; (b) the two membrane protein subunits are initially not associated with each other in the endoplasmic reticulum (ER); (c) heterodimerization occurs predominantly between subunits that originate from the same translation product (heterodimerization in cis); (d) the kinetics of subunit association are very fast (t1/2 = 4 min); and (e) this heterodimerization is highly efficient. To explain the cis-directed heterodimerization reaction we suggest that the p62 protein, which is made before E1 during 26S mRNA translation, is retained at its translocation site until also the E1 chain has been synthesized and translocated at this same site. The mechanism for p62 retention could either be that the p62 anchor sequence cannot diffuse out from an "active" translocation site or that the p62 protein is complexed with a protein folding facilitating machinery that is physically linked to the translocation apparatus.
Assuntos
Vírus da Floresta de Semliki/química , Proteínas Estruturais Virais/química , Animais , Biopolímeros , Linhagem Celular , Cricetinae , Cinética , Testes de Precipitina , Biossíntese de Proteínas , Processamento de Proteína Pós-Traducional , TransfecçãoRESUMO
The envelope of the Semliki Forest virus (SFV) contains two transmembrane proteins, E2 and E1, in a heterodimeric complex. The E2 subunit is initially synthesized as a precursor protein p62, which is proteolytically processed to the mature E2 form before virus budding at the plasma membrane. The p62 (E2) protein mediates binding of the heterodimer to the nucleocapsid during virus budding, whereas E1 carries the entry functions of the virus, that is, cell binding and low pH-mediated membrane fusion activity. We have investigated the significance of the cleavage event for the maturation and entry of the virus. To express SFV with an uncleaved p62 phenotype, BHK-21 cells were transfected by electroporation with infectious viral RNA transcribed from a full-length SFV cDNA clone in which the p62 cleavage site had been changed. The uncleaved p62E1 heterodimer was found to be used for the formation of virus particles with an efficiency comparable to the wild type E2E1 form. However, in contrast to the wild type virus, the mutant virus was virtually noninfectious. Noninfectivity resulted from impaired uptake into cells, as well as from the inability of the virus to promote membrane fusion in the mildly acidic conditions of the endosome. This inability could be reversed by mild trypsin treatment, which converted the viral p62E1 form into the mature E2E1 form, or by treating the virus with a pH 4.5 wash, which in contrast to the more mild pH conditions of endosomes, effectively disrupted the p62E1 subunit association. We conclude that the p62 cleavage is not needed for virus budding, but regulates entry functions of the E1 subunit by controlling the heterodimer stability in acidic conditions.
Assuntos
Fusão de Membrana , Vírus da Floresta de Semliki/fisiologia , Proteínas do Envelope Viral/fisiologia , Proteínas Virais de Fusão/fisiologia , Animais , Linhagem Celular , Cloroquina/farmacologia , Cricetinae , Análise Mutacional de DNA , Endocitose , Imunofluorescência , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Substâncias Macromoleculares , Conformação Proteica , Tripsina/farmacologia , Proteínas do Envelope Viral/ultraestrutura , Proteínas Virais de Fusão/ultraestrutura , Replicação ViralRESUMO
Unlike the wild-type asialoglycoprotein receptor subunit H1 which is transported to the cell surface, endocytosed and recycled, a mutant lacking residues 4-33 of the 40-amino acid cytoplasmic domain was found to be retained intracellularly upon expression in different cell lines. The mutant protein accumulated in the trans-Golgi, as judged from the acquisition of trans-Golgi-specific modifications of the protein and from the immunofluorescence staining pattern. It was localized to juxtanuclear, tubular structures that were also stained by antibodies against galactosyltransferase and gamma-adaptin. The results of further mutagenesis in the cytoplasmic domain indicated that the size rather than the specific sequence of the cytoplasmic domain determines whether H1 is retained in the trans-Golgi or transported to the cell surface. Truncation to less than 17 residues resulted in retention, and extension of a truncated tail by an unrelated sequence restored surface transport. The transmembrane segment of H1 was not sufficient for retention of a reporter molecule and it could be replaced by an artificial apolar sequence without affecting Golgi localization. The cytoplasmic domain thus appears to inhibit interaction(s) of the exoplasmic portion of H1 with trans-Golgi component(s) for example by steric hindrance or by changing the positioning of the protein in the membrane. This mechanism may also be functional in other proteins.
Assuntos
Assialoglicoproteínas/metabolismo , Complexo de Golgi/metabolismo , Proteínas de Membrana/metabolismo , Receptores de Superfície Celular/metabolismo , Sequência de Aminoácidos , Animais , Receptor de Asialoglicoproteína , Sequência de Bases , Linhagem Celular , Retículo Endoplasmático/metabolismo , Imunofluorescência , Humanos , Proteínas de Membrana/química , Proteínas de Membrana/genética , Dados de Sequência Molecular , Mutagênese , Receptores de Superfície Celular/química , Receptores de Superfície Celular/genética , Transfecção , Células Tumorais CultivadasRESUMO
Long-term testosterone replacement therapy is mainly monitored by trough levels of serum testosterone (S-T), while urinary testosterone (U-T) is used by forensic toxicology to evaluate testosterone doping. Testosterone in saliva (Sal-T) may provide additional information and simplify the sample collection. We aimed to investigate the relationships between testosterone measured in saliva, serum and urine during standard treatment with 1,000 mg testosterone undecanoate (TU) every 12th week during 1 year. This was an observational study. Males with primary and secondary hypogonadism (HG; n = 23), subjects with gender dysphoria (GD FtM; n = 15) and a healthy control group of men (n = 32) were investigated. Sal-T, S-T and U-T were measured before and after TU injections. Sal-T was determined with Salimetrics® enzyme immunoassay, S-T with Roche Elecsys® testosterone II assay and U-T by gas chromatography-mass spectrometry. Sal-T correlated significantly with S-T and calculated free testosterone in both controls and patients (HG men and GD FtM), while Sal-T to U-T showed weaker correlations. Trough values of Sal-T after 12 months were significantly higher in the GD FtM group (0.77 ± 0.35 nmol/L) compared to HG men (0.53 ± 0.22 nmol/L) and controls (0.46 ± 0.15 nmol/L), while no differences between S-T and U-T trough values were found. Markedly elevated concentrations of salivary testosterone, 7-14 days after injection, were observed, especially in the GD FtM group. This study demonstrates that Sal-T might be a useful clinical tool to monitor long-term testosterone replacement therapy and might give additional information in forensic cases.
Assuntos
Saliva/química , Testosterona/análise , Adolescente , Adulto , Idoso , Eunuquismo/tratamento farmacológico , Disforia de Gênero/tratamento farmacológico , Terapia de Reposição Hormonal/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Testosterona/uso terapêutico , Adulto JovemRESUMO
The quality of the organs harvested from a deceased donor is of critical importance for the outcome of the transplantations. During 2005, a quality assurance project was initiated to evaluate the donor management, harvest operation, and flow of information during the donation process. Three kinds of questionnaires were used in each donation. They were completed by the transplant coordinator, the harvesting surgeon, and the surgeons performing the liver and kidney transplantations. Feedback is given to the harvesting teams within 2 weeks after the procedures. The most important findings related to missed information concerning organ abnormalities or organ damage from the procurement operation. Procurement of organs from a deceased donor involves a complex chain of events. Based on our experiences in this 1-year project, we believe that standardized registration of the various parts of the process and structured feedback to the staff give possibilities to improve performance. After minor modifications, this method for quality assurance has been introduced as a permanent part of our donation procedure. We believe that this strategy can help to detect weaknesses and improve transplant outcomes.
Assuntos
Obtenção de Tecidos e Órgãos/normas , Atitude Frente a Morte , Família , Humanos , Relações Profissional-Família , Garantia da Qualidade dos Cuidados de Saúde , Inquéritos e Questionários , Suécia , Coleta de Tecidos e Órgãos/métodos , Coleta de Tecidos e Órgãos/normasRESUMO
Since 1990, the Organisation for Organ donation in Central Sweden has registered the numbers of donations at the various hospitals in the area. During this period, a significant decrease in donation rate was observed in the large hospitals, while there was an increase in donation rate in the smaller hospitals. Taken together, the small hospitals are now at least as important as the large hospitals. Possible reasons for the observed change in donation pattern are discussed.
Assuntos
Obtenção de Tecidos e Órgãos/tendências , Número de Leitos em Hospital , Mortalidade Hospitalar , Hospitais de Condado/estatística & dados numéricos , Hospitais de Distrito/estatística & dados numéricos , Hospitais Universitários/estatística & dados numéricos , Humanos , Unidades de Terapia Intensiva , SuéciaRESUMO
Increasing the efficiency of DNA sequencing necessitates the development of systems which reduce the need for manual operations by integrating template preparation, sequencing reactions, product separation and detection. A semi-automated system, whereby PCR-amplified biotinylated genomic or plasmid DNA is immobilized on streptavidin-coated magnetic beads, has been developed.
Assuntos
Sequência de Bases , Automação , Custos e Análise de Custo , DNA , Plasmídeos , Reação em Cadeia da Polimerase/métodosRESUMO
Arteriovenous cross-transfusion and parabiosis were used to study transfer of paraphenylenediamine (PPDA) hypersensitivity in guinea pigs. Nine out of 10 transfusion experiments were successful, resulting in definite positive test reactions in the recipient animals, and the remaining experiment was evaluated as possibly positive. Six out of 10 parabiosis experiments were successful, a further 2 were negative and the remaining 2 were evaluated as possibly positive. The mean test reactions in the recipients were stronger in the transfusion experiments than in the parabiosis experiments (0.001 less than P less than 0.01 for 0.5% PPDA). In control experiments, where the "donors" were nonsensitized guinea pigs negative test reactions were obtained. It was concluded that the transfusion technique was the most reliable of the 2 methods, when studying transfer of PPDA hypersensitivity.
Assuntos
Transfusão de Sangue , Hipersensibilidade Tardia/imunologia , Imunização Passiva , Parabiose , Animais , Transfusão de Sangue/métodos , Hipersensibilidade a Drogas/imunologia , Cobaias , Hipersensibilidade Tardia/induzido quimicamente , Fenilenodiaminas/imunologiaRESUMO
An animal model (the rabbit) was used to define which of 8 chemicals caused pustule formation on topical application. Large occlusive chambers (diameter 12 mm), petrolatum as the vehicle and wrapping contributed to efficient occlusion and pustulation. Sodium lauryl sulfate and mecuric chloride gave reproducible results and clear dose-responses indicating that this pustulation is an expression of primary irritancy. Ammonium fluoride pustulation was not reproducible; croton oil pustules were more difficult to evaluate due to simultaneous erythema and edema. Sodium arsentate, nickel sulfate and potassium iodide pustules developed at sites where the skin barriers had been damaged by a stab injury. Benzalkonium chloride caused yellow staining and edema but not pustules. Because of lack of epidemiologic data, we do not know how frequently similar findings occur in man.
Assuntos
Modelos Animais de Doenças , Irritantes , Dermatopatias/induzido quimicamente , Compostos de Amônio , Animais , Arseniatos/toxicidade , Compostos de Benzalcônio/toxicidade , Óleo de Cróton/toxicidade , Relação Dose-Resposta a Droga , Fluoretos/toxicidade , Cloreto de Mercúrio , Mercúrio/toxicidade , Níquel/toxicidade , Iodeto de Potássio/toxicidade , Compostos de Amônio Quaternário/toxicidade , Coelhos , Pele/efeitos dos fármacos , Dodecilsulfato de Sódio/toxicidade , Supuração/induzido quimicamenteRESUMO
The purpose of this study was to evaluate the value of additive components and colloid included in the University of Wisconsin (UW) solution. Therefore, this solution was compared with a solution consisting of the basic components of the UW solution (potassium lactobionate, raffinose, phosphate buffer and MgSO4). We employed a method of measuring the amount of chromium-51-labeled erythrocyte trapped in the medullary vasculature 20 min after reperfusion of kidney grafts cold-stored for 24-48 h in either the basic UW (bUW) or the original UW (oUW) solution. The amount of trapping has been shown to correlate well with the degree of cold ischemic injury. Both hemodiluted (hct 20-27%) and normal (hct 41-45%) recipients were used. Long-term viability of grafts stored in either bUW or oUW was investigated in survival experiments and the flow rates during in situ flush-out were also measured, as well as weight changes during the storage period. The results showed no significant difference between the two solutions, regardless of ischemia time or whether hemodiluted or normal recipients were used. However, the flow rate and weight measurements showed that flushing was more rapid and kidney swelling less pronounced using oUW. Survival rates in long-term transplantation experiments were similar. It was concluded that the inclusion of a colloid improves the rheological properties of the UW solution and that the additives besides the basic components did not offer any advantage.
Assuntos
Rim , Preservação de Órgãos/métodos , Animais , Soluções Tampão , Temperatura Baixa , Hematócrito , Isquemia , Rim/irrigação sanguínea , Masculino , Ratos , Ratos Endogâmicos , Fatores de TempoRESUMO
The role of recipient hemodilution for postischemic renal medullary red cell trapping was investigated after different periods of cold storage in a conventional cold storage solution (Sacks'). At all cold storage times investigated (4, 12, 24, and 48 hr) medullary red cell trapping was reduced by isovolemic hemodilution, with about 50% reduction of recipient hematocrit. Trapping was also reduced when a modification of a new preservation solution (University of Wisconsin solution [UW]) was used and compared with flush-out and storage in a standard preservation solution (Sacks'). The combination of hemodilution and preservation in modified UW solution had additional capacity to reduce medullary red cell trapping. Thus, even after 48 hr of cold storage, only a moderate trapping was observed. The results also indicate that measurements of medullary red cell trapping offers an accurate method of grading postischemic renal damage.
Assuntos
Eritrócitos/metabolismo , Hemodiluição , Isquemia/complicações , Transplante de Rim , Soluções para Preservação de Órgãos , Preservação de Órgãos , Adenosina , Alopurinol , Animais , Temperatura Baixa , Glutationa , Insulina , Rim/irrigação sanguínea , Rim/fisiologia , Masculino , Preservação de Órgãos/métodos , Rafinose , Ratos , Ratos Endogâmicos , SoluçõesRESUMO
Inbred male Sprague-Dawley rats were transplanted with syngeneic pancreaticoduodenal grafts. Diabetes was induced in some of the recipient animals by an i.v. injection of streptozotocin one week before transplantation, while the remaining control rats were untreated, and thus had both an intact native and a transplanted pancreas. The left kidney of the recipient was removed and its blood vessels were used for the graft vascular anastomosis. The graft duodenum was sutured end-to-side to the ileum of the recipient. Two weeks after transplantation the rates of blood flow through both the native and transplanted pancreas and duodenum were measured. The blood perfusion of the whole pancreas and the islets was higher in the tx than in the native gland, although no difference was seen with regard to the duodenum. Administration of glucose to control animals had no effect on either native or tx whole pancreatic or intestinal blood flow. However, the islet blood flow through the native pancreas was increased in response to glucose, while that of the tx gland remained unchanged. The combined data show that the blood flow of a tx pancreas is slightly higher than that of the native organ, and that the response of the islet blood perfusion to glucose administration differs between the native and tx pancreas. This suggests that islets in the tx pancreas 14 days after transplantation are not subject to the same blood flow regulatory mechanisms as native islets.
Assuntos
Duodeno/transplante , Ilhotas Pancreáticas/irrigação sanguínea , Transplante de Pâncreas , Animais , Glicemia/análise , Duodeno/irrigação sanguínea , Insulina/sangue , Masculino , Ratos , Ratos Endogâmicos , Fluxo Sanguíneo RegionalRESUMO
A new flushout solution for preservation of the pancreas was tested in the dog model of segmental pancreas autotransplantation. The solution osmolality was 320 mOsm/L, K+ = 120 mM, Na+ = 30 mM, and it contained the anion, lactobionate, and raffinose as impermeants to the cell. Preservation times studied were 48 and 72 hr. The pancreas was flushed out with about 250 ml of the new solution and stored at 0 degrees C. Dogs were monitored postoperatively for blood glucose and intravenous glucose tolerance (IVGTT). Results were compared with control (no preservation) segmental pancreas autotransplants. Dogs receiving pancreases stored for 48 or 72 hr were normoglycemic on day one and remained normoglycemic for at least 28 days, or until time of sacrifice. Two of four dogs with pancreases stored for 48 hr were sacrificed on day 14 with normal IVGTT for histology. The remaining two dogs had normal pancreatic function for 28 days. Two of eight dogs receiving pancreas grafts after 72-hr cold storage died of causes unrelated to the pancreas graft, which was still functioning at the time of death. Six dogs remained normoglycemic and had a normal IVGTT at least for 28 days. This study demonstrates the feasibility of preserving the pancreas for three days for transplantation.