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1.
Tech Coloproctol ; 27(1): 75-81, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36029385

RESUMO

The management of low rectal cancer is a perennial challenge for colorectal surgeons. The benefits of transanal total mesorectal excision (TaTME) in low rectal cancer are to secure the distal margin and avoid surgical space constraints within the deep pelvis. However, anastomotic leak remains an important concern. We report our technique and results combining TaTME with delayed coloanal anastomosis (DCAA) without bowel diversion. First, the splenic flexure, left colon and rectum are laparoscopically mobilized to mid-rectum. TaTME is performed to complete the distal rectal mobilization, and the specimen is delivered transanally and transected. The abdominoperineal colonic pull-through is secured to the anal canal and hypertonic dressing is applied regularly in the ward. The handsewn DCAA is performed one week later. An accompanying video demonstrates this technique. Five consecutive patients with low rectal cancer underwent TaTME with DCAA. All had upfront surgical resection except one who underwent total neoadjuvant therapy. Mean operative duration, blood loss, and length of hospital stay was 290 (250-375) min, 142 (10-200) ml and 11.6 (10-14) days respectively. One patient (20%) suffered a postoperative complication of persistent urinary retention, requiring an indwelling urinary catheter on discharge. There were no cases of open conversion and no instances of anastomotic leakage. Two patients (40%) had minor low anterior resection syndrome (LARS) and one (20%) had major LARS. TaTME and DCAA without stoma are complimentary techniques that augment the minimally invasive effects of laparoscopic sphincter-sparing low rectal cancer surgery, with good perioperative outcomes.


Assuntos
Laparoscopia , Neoplasias Retais , Cirurgia Endoscópica Transanal , Humanos , Neoplasias Retais/cirurgia , Canal Anal/cirurgia , Complicações Pós-Operatórias/etiologia , Complicações Pós-Operatórias/cirurgia , Tratamentos com Preservação do Órgão , Reto/cirurgia , Anastomose Cirúrgica/métodos , Laparoscopia/métodos , Fístula Anastomótica/etiologia , Fístula Anastomótica/cirurgia , Síndrome de Ressecção Anterior Baixa , Cirurgia Endoscópica Transanal/métodos , Resultado do Tratamento
2.
Hong Kong Med J ; 28(3): 215-222, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35686472

RESUMO

INTRODUCTION: Compared with previous waves of the coronavirus disease 2019 (COVID-19) pandemic in Hong Kong, the third wave involved a greater number of frail older patients. Because local healthcare policy required hospitalisation for all older adults with COVID-19, we aimed to investigate the clinical course and outcomes in such patients. METHODS: This retrospective observational study included all patients aged ≥65 years who were admitted to Tuen Mun Hospital for management of COVID-19 between 1 July 2020 and 31 August 2020. We reviewed baseline characteristics, clinical presentation, laboratory results, complications, and outcomes. We also investigated the associations of age and Clinical Frailty Scale (CFS) score with in-patient mortality. RESULTS: In total, 101 patients were included (median age, 73 years); 52.5% were men and 85% had at least co-morbid chronic disease. The most common symptoms were fever (80.2%) and cough (63.4%). Fifty-two patients (51.5%) developed hypoxia, generally on day 8 (interquartile range, 5-11) after symptom onset. Of the 16 patients who required intensive care unit support, 13 required mechanical ventilation. The overall mortality rate was 16.8%. Patients aged 65-69, 70-79, 80-89, and ≥90 years had mortality rates of 9.1%, 10%, 30%, and 25%, respectively. Patients with CFS scores of 1-2, 3-4, 5-6, and ≥7 had mortality rates of 5.7%, 14.7%, 23.5%, and 40%, respectively. A linear relationship was confirmed between the two mortality trends. CONCLUSION: Clinical deterioration was common in older patients with COVID-19; their overall mortality rate was 16.8%. Mortality increased linearly with both age and CFS score.


Assuntos
COVID-19 , Fragilidade , Idoso , COVID-19/terapia , Feminino , Hong Kong/epidemiologia , Mortalidade Hospitalar , Humanos , Masculino , Estudos Observacionais como Assunto , Pandemias
3.
Gastroenterology ; 159(1): 81-95, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32251668

RESUMO

BACKGROUND & AIMS: Infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19), which has been characterized by fever, respiratory, and gastrointestinal symptoms as well as shedding of virus RNA into feces. We performed a systematic review and meta-analysis of published gastrointestinal symptoms and detection of virus in stool and also summarized data from a cohort of patients with COVID-19 in Hong Kong. METHODS: We collected data from the cohort of patients with COVID-19 in Hong Kong (N = 59; diagnosis from February 2 through February 29, 2020),and searched PubMed, Embase, Cochrane, and 3 Chinese databases through March 11, 2020, according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. We analyzed pooled data on the prevalence of overall and individual gastrointestinal symptoms (loss of appetite, nausea, vomiting, diarrhea, and abdominal pain or discomfort) using a random effects model. RESULTS: Among the 59 patients with COVID-19 in Hong Kong, 15 patients (25.4%) had gastrointestinal symptoms, and 9 patients (15.3%) had stool that tested positive for virus RNA. Stool viral RNA was detected in 38.5% and 8.7% among those with and without diarrhea, respectively (P = .02). The median fecal viral load was 5.1 log10 copies per milliliter in patients with diarrhea vs 3.9 log10 copies per milliliter in patients without diarrhea (P = .06). In a meta-analysis of 60 studies comprising 4243 patients, the pooled prevalence of all gastrointestinal symptoms was 17.6% (95% confidence interval [CI], 12.3-24.5); 11.8% of patients with nonsevere COVID-19 had gastrointestinal symptoms (95% CI, 4.1-29.1), and 17.1% of patients with severe COVID-19 had gastrointestinal symptoms (95% CI, 6.9-36.7). In the meta-analysis, the pooled prevalence of stool samples that were positive for virus RNA was 48.1% (95% CI, 38.3-57.9); of these samples, 70.3% of those collected after loss of virus from respiratory specimens tested positive for the virus (95% CI, 49.6-85.1). CONCLUSIONS: In an analysis of data from the Hong Kong cohort of patients with COVID-19 and a meta-analysis of findings from publications, we found that 17.6% of patients with COVID-19 had gastrointestinal symptoms. Virus RNA was detected in stool samples from 48.1% patients, even in stool collected after respiratory samples had negative test results. Health care workers should therefore exercise caution in collecting fecal samples or performing endoscopic procedures in patients with COVID-19, even during patient recovery.


Assuntos
Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/prevenção & controle , Diarreia/virologia , Fezes/virologia , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , Carga Viral , Betacoronavirus/genética , Betacoronavirus/patogenicidade , COVID-19 , Teste para COVID-19 , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/transmissão , Diarreia/diagnóstico , Diarreia/epidemiologia , Endoscopia Gastrointestinal/normas , Trato Gastrointestinal/diagnóstico por imagem , Trato Gastrointestinal/virologia , Hong Kong/epidemiologia , Humanos , Controle de Infecções/normas , Transmissão de Doença Infecciosa do Paciente para o Profissional/prevenção & controle , Pneumonia Viral/diagnóstico , Pneumonia Viral/epidemiologia , Pneumonia Viral/transmissão , Prevalência , RNA Viral/isolamento & purificação , SARS-CoV-2
4.
J Clin Pharm Ther ; 40(2): 240-4, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25487647

RESUMO

WHAT IS KNOWN AND OBJECTIVE: Dosage adjustment of 500 mg ertapenem daily is recommended for patients with advanced kidney disease. 30% of ertapenem is cleared by a session of haemodialysis (HD). However, because most published carbapenems studies have excluded patients on dialysis, little is known about the dosing of ertapenem to avoid central nervous system (CNS) toxicity in regular HD patients. We report of four patients who developed CNS toxicity in such patients. CASE SUMMARY: The 4 HD patients developed unexplained CNS toxicity manifested as seizures, hallucination and cognitive dysfunction after receiving 3-7 consecutive recommended doses of ertapenem. Their symptoms of CNS toxicity were completely resolved within 8 days after discontinuation of ertapenem. In one of our presented cases, we demonstrated the very high level of plasma ertapenem accumulating with several consecutive doses. Cognitive function gradually recovered in line with a corresponding decline in blood level of ertapenem. WHAT IS NEW AND CONCLUSIONS: This is the first report of ertapenem-associated CNS toxicity in patients on regular HD and utilizing the plasma ertapenem concentration to demonstrate the causal relationship. The recommended dosage of 500 mg ertapenem daily may be still too high in regular HD patients, especially in Asians, owing to their relatively small body size. An increased awareness of ertapenem-associated CNS toxicity would avoid unnecessary examinations, hospitalization, and potentially catastrophic complications.


Assuntos
Antibacterianos/efeitos adversos , Antibacterianos/farmacocinética , Doenças do Sistema Nervoso Central/induzido quimicamente , Diálise Renal , beta-Lactamas/efeitos adversos , beta-Lactamas/farmacocinética , Idoso , Povo Asiático , Relação Dose-Resposta a Droga , Ertapenem , Feminino , Humanos , Masculino , Insuficiência Renal
5.
Front Allergy ; 3: 974138, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36133403

RESUMO

Introduction: Penicillin allergy testing has been traditionally performed by allergists, but there remains a huge deficit of specialists. A multidisciplinary effort with nonallergists would be invaluable to overcome the magnitude of penicillin allergy labels via the Hong Kong Drug Allergy Delabelling Initiative (HK-DADI). These consensus statements (CSs) offer recommendations and guidance to enable nonallergists to screen for low-risk (LR) patients and perform penicillin allergy testing. Methods: CSs were formulated by the HK-DADI Group using the Delphi method. An agreement was defined as greater than or equal to 80% consensus. Results: A total of 26 CSs reached consensus after multiple rounds of Delphi. CSs were categorized into risk assessment, skin testing, drug provocation testing (DPT), and post-testing management. For risk assessment, the essentials of allergy history and exclusion criteria were detailed. Patients with only LR features can proceed with testing by nonallergists. Skin tests should be performed prior to DPT. Details regarding the timing, preparation, and interpretation of skin tests were elaborated. DPT remains the gold standard to diagnose genuine allergy or tolerance and should be performed when there is a low pretest probability following negative skin testing. Details of DPT preparations, dosing protocols, and interpretation were elaborated. For post-testing management, inaccurate allergy labels should be delabeled following negative DPT with proper patient counseling. Conclusion: CSs support penicillin allergy testing by nonallergists in Hong Kong. LR cases can be managed by nonallergists at Spoke Clinics, with training and support of an allergist-led Hub.

6.
Nat Protoc ; 16(10): 4650-4675, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34400840

RESUMO

Ca2+ handling within cardiac myocytes underpins coordinated contractile function within the beating heart. This protocol enables high spatial and temporal Ca2+ imaging of ex vivo multicellular myocardial strips. The endocardial surface is retained, and strips of 150-300-µm thickness are dissected, loaded with Ca2+ indicators and mounted within 1.5 h. A list of the equipment and reagents used and the key methodological aspects allowing the use of this technique on strips from any chamber of the mammalian heart are described. We have successfully used this protocol on human, pig and rat biopsy samples. On use of this protocol with intact endocardial endothelium, we demonstrated that the myocytes develop asynchronous spontaneous Ca2+ events, which can be ablated by electrically evoked Ca2+ transients, and subsequently redevelop spontaneously after cessation of stimulation. This protocol thus offers a rapid and reliable method for studying the Ca2+ signaling underpinning cardiomyocyte contraction, in both healthy and diseased tissue.


Assuntos
Sinalização do Cálcio , Miocárdio , Miócitos Cardíacos , Animais , Contração Miocárdica , Ratos , Suínos
7.
Artigo em Inglês | MEDLINE | ID: mdl-34527820

RESUMO

BACKGROUND: Weekly toxicity assessments for patients undergoing head and neck (HN) radiotherapy are essential to ensure that acute side effects are appropriately managed in order for patients to complete their treatment in a safe and timely manner. The incorporation of Advanced Practice Radiation Therapist (APRT) led treatment reviews has been reported for various subsites, but there is currently a lack of published literature regarding this role for patients with HN cancer. The purpose of this study is to assess the concordance of toxicity assessments performed during weekly radiotherapy treatment reviews for patients undergoing HN radiotherapy between the HN APRT and Radiation Oncologist (RO). METHODS: Twenty-three patients with nasopharyngeal cancer (NPC) under the care of 3 ROs were recruited from June to December 2018; weekly assessments were independently performed by HN APRT and ROs. The HN toxicity assessment was graded according to the Common Terminology Criteria for Advanced Events v4.0. Both assessors were blinded to each other's assessments. The percentage agreement of concordance and agreement level were interpreted by Cohen's Kappa statistic (κ), with the ROs' assessments deemed as the 'gold standard'. RESULTS: The overall concordance for all graded toxicity assessments between HN APRT and ROs was 78.4%. Xerostomia, dysgeusia, pharyngeal pain and dermatitis assessment were evaluated as 'Good' with agreement ranging from κ = 0.608-0.640 between the HN APRT and ROs while dysphagia scored an 'Almost Perfect' agreement of κ = 0.834. 'Moderate' agreement between the HN APRT and ROs was observed for oral pain and mucositis assessment. A scoring discrepancy of 1 and 2 grades was observed in 21.2% and 0.4% for these two toxicities. CONCLUSION:  There was high concordance in scoring of acute toxicity between the HN APRT and ROs. The results support the continuing involvement of HN APRT in weekly assessments for NPC patients.

9.
Resuscitation ; 149: 39-46, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32027981

RESUMO

BACKGROUND: Our study aimed to identify a strategy that maximizes survival upon hospital discharge or 30-days post out-of-hospital cardiac arrest (OHCA) in Singapore for fixed investments of S$1, S$5, or S$10 million. Four strategies were compared: (1) no additional investment; (2) reducing response time via leasing of more ambulances; (3) increasing number of people trained in cardiopulmonary resuscitation (CPR); and (4) automated external defibrillators (AED). METHODS: We estimated the effect of ambulance response time, bystander CPR and AED on survival based on Singapore's 2010-2015 OHCA registry data. We simulated the changes in ambulance response times and likelihood of (1) CPR and (2) AED usage as a function of their increased availability, which was then combined with the effect of each factor to determine the increase in survival for each strategy. RESULTS: Survival given no additional investment was 4.03% (95% CI: 3.96%, 4.10%). The investments in ambulances, CPR training and AEDs for a given budget of S$1M changed survival to 4.03% (95% CI: 3.96%, 4.10%), 4.04% (95% CI: 3.98%, 4.11%), and 4.44% (95% CI: 4.35%, 4.54%), respectively. This generated 0, 2 and 102 additional life years saved respectively. Given a budget of S$5M or S$10M, 509 or 886 additional life years could be saved, by investing in an additional 10,000 or 20,000 AEDs respectively. The strategies reached a saturation effect whereby improvement in survival was marginal when the budget was increased to ≥S$5M for investment in ambulances and CPR training. CONCLUSIONS: Investing in AEDs had the most gain in survival.


Assuntos
Reanimação Cardiopulmonar , Serviços Médicos de Emergência , Parada Cardíaca Extra-Hospitalar , Desfibriladores , Humanos , Parada Cardíaca Extra-Hospitalar/terapia , Singapura/epidemiologia
10.
J Appl Microbiol ; 106(4): 1268-79, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19187146

RESUMO

AIMS: Preconditioning of stainless steel with aqueous cod muscle extract significantly impedes subsequent bacterial adhesion most likely due to repelling effects of fish tropomyosin. The purpose of this study was to determine if other food conditioning films decrease or enhance bacterial adhesion to stainless steel. METHODS AND RESULTS: Attachment of Pseudomonas fluorescens AH2 to stainless steel coated with water-soluble coatings of animal origin was significantly reduced as compared with noncoated stainless steel or stainless steel coated with laboratory substrate or extracts of plant origin. Coating with animal extracts also decreases adhesion of other food-relevant bacteria. The manipulation of adhesion was not attributable to growth inhibitory effects. Chemical analysis revealed that the stainless steels were covered by homogenous layers of adsorbed proteins. The presence of tropomyocin was indicated by appearance of proteins with similar molecular weight based in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, in several extracts that reduced adhesion but also extracts not containing this protein reduced bacterial adhesion, indicating that several molecular species may be involved in the phenomenon. CONCLUSIONS: It is a common perception that food materials facilitate bacterial adhesion to surfaces; however, this study demonstrates that aqueous coatings of food origin may actually reduce bacterial adhesion. SIGNIFICANCE AND IMPACT OF THE STUDY: Compounds from food extracts may potentially be used as nontoxic coatings to reduce bacterial attachment to inert surfaces.


Assuntos
Antibacterianos/farmacologia , Bactérias/crescimento & desenvolvimento , Aderência Bacteriana/fisiologia , Biofilmes/crescimento & desenvolvimento , Microbiologia de Alimentos , Proteínas/farmacologia , Aço Inoxidável , Animais , Contagem de Colônia Microbiana , Eletroforese em Gel de Poliacrilamida , Peixes , Extratos Vegetais/farmacologia , Pseudomonas fluorescens/crescimento & desenvolvimento , Aço Inoxidável/química , Propriedades de Superfície , Tropomiosina/análise , Espectroscopia por Absorção de Raios X
11.
Resuscitation ; 132: 85-89, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30171975

RESUMO

INTRODUCTION: The Global Resuscitation Alliance (GRA) was established in 2015 to improve survival for Out- of-Hospital Cardiac Arrest (OHCA) using the best practices developed by the Seattle Resuscitation Academy. However, these 10 programs were recommended in the context of developed Emergency Care Systems (ECS). Implementing these programs can be challenging for ECS at earlier stages of development. We aimed to explore barriers faced by developing ECS and to establish pre-requisites needed. We also developed a framework by which developing ECS may use to build their emergency response capability. METHOD: A consensus meeting was held in Singapore on 1st-2nd August 2017. The 74 participants were key stakeholders from 26 countries, including Emergency Medical Services (EMS) directors, physicians and academics, and two Physicians who sit on the World Health Organisation (WHO) panel for development of Emergency Care Systems. Five discussion groups examined the chain of survival: community, dispatch, ambulance and hospital; a separate group considered perinatal resuscitation. Discussion points were voted upon to reach a consensus. RESULTS: The answers and discussion points from each groupwere classified into a table adapted from WHO's framework of development for Emergency Services. After which, it was used to construct the modified survival framework with the chain of survival as the backbone. Eleven key statements were then derived to describe the pre-requisites for achieving the GRA 10 programs. The participants eventually voted on the importance and feasibility of these 11 statements as well as the GRA 10 programs using a matrix that is used by organisations to prioritise their action steps. CONCLUSION: In this paper, we propose a modified framework of survival for developing ECS systems. There are barriers for developing ECS systems to improve OHCA survival rates. These barriers may be overcome by systematic prioritisation and cost-effective innovative solutions.


Assuntos
Serviços Médicos de Emergência/organização & administração , Parada Cardíaca Extra-Hospitalar/mortalidade , Reanimação Cardiopulmonar/normas , Participação da Comunidade , Conferências de Consenso como Assunto , Saúde Global , Humanos , Parada Cardíaca Extra-Hospitalar/terapia
12.
Ann Acad Med Singap ; 34(9): 586-90, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16284685

RESUMO

The Singapore Humanitarian Assistance Support Group deployed a team of 32 medical relief workers to Meulaboh, Indonesia to provide medical assistance for victims of the 26 December earthquake and tsunami disaster. The team was deployed at a primary healthcare clinic at an internally displaced persons' (IDP) camp and at the sole hospital's emergency and surgical departments. The team saw a total of 1841 patients, 1371 at the clinic and 446 at the hospital's emergency department, and performed surgery on 24 patients. Tsunami-related trauma cases accounted for 31.8% (142) of cases at the emergency department, 1.6% (22) of cases at the clinic, and 91.7% (22) of surgeries. This paper details the difficulties and lessons learnt by the team, including the lack of important resources for healthcare delivery. Water, sanitation, hygiene, and vector control were some of the problems faced, with the goal to provide the most effective public health for the greatest number of people given the limited resources available.


Assuntos
Desastres , Socorro em Desastres , Humanos , Indonésia , Socorro em Desastres/organização & administração , Socorro em Desastres/estatística & dados numéricos
13.
Ann Acad Med Singap ; 34(9): 539-4, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16284674

RESUMO

INTRODUCTION: A common approach to the surveillance of emerging infectious diseases and agents of bioterrorism is to analyse electronically captured data for disease syndromes. The Patient Care Enhancement System (PACES) is a form of electronic medical records presently in service in the Singapore Armed Forces (SAF). We assess the feasibility of PACES data for surveillance, describe time-trends, and identify methods of sub-analysis which could improve performance. MATERIALS AND METHODS: Medical consults from July 2000 to June 2003 were extracted. Diagnosis codes were mapped to 7 infectious disease syndromes according to the categorisation in the Electronic Surveillance System for the Early Notification of Community-based Epidemics (ESSENCE): gastrointestinal (GI), fever (FEVER), respiratory, (RESP), coma (COMA), neurological (NEURO), dermatologic-haemorrhagic (DERMHEM) and dermatologic- infectious (DERM-INF). RESULTS: A total of 732,233 episodes of care were analysed. Weekly periodicity was observed, with decreased weekend consults; there were no obvious seasonal trends in any of the syndromes. RESP, FEVER and GI syndromes were common events. Sub-analyses, either by restricting to cases with a repeated consultation, or grouping the data by medical centres, could dramatically lower thresholds used to flag outbreaks. CONCLUSION: In spite of the level of background noise inherent in a system consisting mainly of primary care consults, sub-analysis by medical centre, or restriction to cases with repeated consults were able to yield sensitive thresholds for outbreak detection.


Assuntos
Infecções/epidemiologia , Sistemas Computadorizados de Registros Médicos , Adolescente , Adulto , Humanos , Vigilância da População , Síndrome
14.
Cancer Gene Ther ; 7(4): 624-8, 2000 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10811481

RESUMO

A major goal in retroviral-based gene therapy is to establish methods that allow for the selection and tracking of transduced cell populations. Ex vivo gene marking of normal and malignant hemopoietic cells allows the cells to be followed subsequently in vivo. For in vivo applications, a neutral marker gene that is nonimmunogenic is desirable. To track two distinctively treated cell populations in a single individual, we designed and constructed two retroviral vectors; both of these vectors encode a truncated form of the human low-affinity nerve growth factor receptor, a neutral gene that does not transduce signals and is expected to be nonimmunogenic in humans. The two vectors, named Frape-1 and Frape-3, are identical at the protein level but differ at the DNA level, containing restriction sites that allow easy detection by polymerase chain reaction analysis. We show that cell lines and primary CD34+ cells can be readily transduced with these vectors and that transduced cells can be distinguished by polymerase chain reaction- and vector-specific restriction sites. These vectors will be useful for toxicity studies on in vivo gene therapy and for determining the source of relapse in hematological malignancies.


Assuntos
Vetores Genéticos , Células-Tronco Hematopoéticas/citologia , Receptores de Fator de Crescimento Neural/genética , Retroviridae/genética , Transfecção/métodos , Antígenos CD34 , Células da Medula Óssea/citologia , Células Cultivadas , Marcadores Genéticos , Células-Tronco Hematopoéticas/efeitos dos fármacos , Células-Tronco Hematopoéticas/fisiologia , Humanos , Células Jurkat , Proteínas de Membrana/farmacologia , Mieloma Múltiplo , Receptores de Fator de Crescimento Neural/análise , Mapeamento por Restrição , Fator de Células-Tronco/farmacologia , Trombopoetina/farmacologia , Células Tumorais Cultivadas
15.
J Histochem Cytochem ; 44(3): 281-7, 1996 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-8648089

RESUMO

The digestion of fixed tissue sections is a critical step in the optimization of any in situ hybridization protocol. We describe a novel application of microwave oven heating to optimize mRNA detection in paraformaldehyde-fixed tissues by in situ hybridization using digoxigenin-labeled probes. This technique replaces protease digestion of fixed tissue sections with 10 min of microwave pretreatment, followed by either conventional hybridization or hybridization involving microwave incubation. This new technique has several advantages over the standard protease treatment-based methods presently in use. (a) Microwave oven heating is a simple, rapid, and highly reproducible technique. (b) Microwave pretreatment significantly increased the hybridization signal and reduced the background compared to conventional protease digestion. Consequently, the hybridization time required to obtain optimal mRNA detection was reduced to 30 min. (c) Ten minutes of microwave pretreatment produced an optimal hybridization signal in six different tissues using a variety of probes, demonstrating the general applicability of this technique. (d) Microwave heating of the probe during the hybridization step itself further reduced the hybridization time and substantially enhanced the hybridization signal obtained from proteinase K-digested tissue. (e) Microwave pretreatment caused no discernible loss of fine cell structure and tissue morphology compared to untreated tissue sections. In conclusion, microwave oven heating can replace the complicated strategies and poor reproducibility of protease treatment of tissue sections, resulting in a simple, rapid, more reliable and sensitive method that has general applicability for in situ hybridization.


Assuntos
Hibridização In Situ/métodos , RNA Mensageiro/análise , Animais , Temperatura Alta , Micro-Ondas , Ratos , Sensibilidade e Especificidade , Inclusão do Tecido , Fixação de Tecidos
16.
Am J Kidney Dis ; 37(4): 820-31, 2001 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-11273883

RESUMO

Interleukin-1 (IL-1) has been shown to exert profibrotic activity in a number of disease models, including crescentic glomerulonephritis and pulmonary fibrosis, but the mechanisms by which this operates are poorly understood. Recent studies have identified a novel mechanism promoting renal fibrosis: tubular epithelial-myofibroblast transdifferentiation (TEMT). The present study examined whether IL-1 can stimulate TEMT in vitro. Cells of the normal rat kidney tubular epithelial cell line (NRK52E) were grown to confluence on collagen-coated plates and cultured for 5 days in the presence 1 to 20 ng/mL of IL-1alpha. Doses of 10 to 20 ng/mL of IL-1 caused transdifferentiation of NRK52E cells into myofibroblast-like cells. Scanning electron microscopy identified IL-1-induced morphological changes as a loss of apical-basal polarity and microvilli, cell hypertrophy, and the development of an elongated and invasive appearance. Phenotypically, IL-1-induced TEMT was characterized by de novo messenger RNA and protein expression of the mesenchymal marker alpha-smooth muscle actin, shown by Northern blotting, immunohistochemistry, and Western blotting. This was accompanied by loss of the epithelial marker E-cadherin. The addition of an excess of IL-1-receptor antagonist completely inhibited IL-1-induced TEMT. IL-1 was shown to stimulate the secretion of active transforming growth factor-beta1 (TGF-beta1) by NRK52E cells. Furthermore, the addition of a neutralizing anti-TGF-beta1 antibody inhibited IL-1-induced TEMT. In conclusion, IL-1 is a profibrogenic cytokine capable of inducing TEMT through a TGF-beta1-dependent mechanism. This may represent a novel mechanism by which IL-1 induces renal fibrosis in vivo.


Assuntos
Fibrose/fisiopatologia , Interleucina-1/fisiologia , Nefropatias/fisiopatologia , Túbulos Renais/fisiologia , Actinas/biossíntese , Actinas/metabolismo , Análise de Variância , Animais , Northern Blotting , Western Blotting , Caderinas/biossíntese , Caderinas/metabolismo , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Linhagem Celular , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/fisiologia , Fibrose/etiologia , Imuno-Histoquímica , Interleucina-1/farmacologia , Nefropatias/etiologia , Falência Renal Crônica/etiologia , Falência Renal Crônica/fisiopatologia , Túbulos Renais/citologia , Microscopia Eletrônica , Músculo Liso/química , Ratos
17.
Resuscitation ; 10(1): 13-8, 1982 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-6291114

RESUMO

Transcutaneous carbon dioxide tensions (tcPCO2) measured with a heated electrode were compared with arterial carbon tensions (PaCO2) in 28 patients. Seventy-eight observations were made. At a skin electrode temperature of 44 degrees C the tcPCO2 was significantly higher than PaCO2 but changes in PCO2 detected by both measurements were closely correlated. The correlation coefficient between tcPCO2 and PaCO2 was 0.92 (P less than 0.001). The 90% response time of the electrode in vitro was less than 1 min, and in vivo stabilization of the recording occurred in less than 15 min. It was found that the transcutaneous PCO2 electrode can be employed usefully in intensive care monitoring of adult patients especially when weaning them from artificial ventilation.


Assuntos
Dióxido de Carbono/sangue , Cuidados Críticos/métodos , Monitorização Fisiológica/métodos , Adulto , Idoso , Eletrodos , Humanos , Pessoa de Meia-Idade , Pressão Parcial , Pele
18.
Clin Nephrol ; 54(2): 157-60, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10968694

RESUMO

We report an unusual case of inappropriate antidiuresis with undetectable vasopressin in an elderly man presenting with confusion due to severe hyponatremia. Further investigations led to the diagnosis of non-functional pituitary macroadenoma. The patient had normal thyroid and adrenal function. The abnormal water balance resolved promptly after transsphenoidal removal of the tumor, confirmed by a repeat water loading test. We conclude that inappropriate antidiuresis in the absence of excess vasopressin secretion may implicate mass effect from an underlying pituitary tumor.


Assuntos
Adenoma/complicações , Síndrome de Secreção Inadequada de HAD/etiologia , Neoplasias Hipofisárias/complicações , Vasopressinas/metabolismo , Adenoma/fisiopatologia , Idoso , Confusão/etiologia , Humanos , Hiponatremia/etiologia , Síndrome de Secreção Inadequada de HAD/fisiopatologia , Masculino , Neoplasias Hipofisárias/fisiopatologia
19.
Clin Nephrol ; 57(4): 289-95, 2002 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-12005245

RESUMO

AIMS: To investigate the relation of leukopenia and thrombocytopenia in hemodialysis (HD) patients with hepatitis C virus (HCV) infection. MATERIALS AND METHODS: The study included 86 HD patients with hepatitis B surface antigen-negative and hepatitis C antibody-negative, 28 HD patients with hepatitis C antibody-positive, 22 HD patients with hepatitis B surface antigen-positive, 78 non-HD patients with hepatitis B-induced liver cirrhosis and 38 non-hemodialysis patients with hepatitis C-induced liver cirrhosis. The following parameters were checked: anti-HCV, hepatitis B surface antigen, hemoglobin, hematocrit, white blood cells, platelets, calcium, phosphate, iron, ferritin, albumin, globulin, aspartate transaminase (AST), alanine transaminase (ALT) and C-reactive protein. The history of blood transfusions, medications, erythropoietin doses and adequate dialysis (KTNV) for 6 consecutive months was also recorded from charts. RESULTS: The HD patients with positive serum anti-HCV and non-HD patients with hepatitis B- or C-induced liver cirrhosis had higher prevalences of leukopenia (39.3%, 43.6% and 50% vs. 15.1%; p < 0.001) and thrombocytopenia (67.9%, 89.7% and 81.6% vs. 34.9%: p < 0.001) than HD patients with serum anti-HCV(-)HbsAg(-). The WBC (4,432 +/- 1,394, 4,792 +/- 2,263 and 4,624 2,446 vs. 5,590 +/- 1,500/mm3; p < 0.001) and platelet counts (140 +/- 45, 80 +/- 50 and 89 +/- 65 vs. 186 +/- 62 x 10(3)/mm3; p < 0.001) of HD patients with positive serum anti-HCV and non-HD patients with hepatitis B- or C-induced cirrhosis were also lower than HD patients without anti-HCV antibody. The liver cirrhosis patients had more thrombocytopenia than the HD patients with anti-HCV(+). The WBC and platelet counts did not vary between HD patients with HbsAg(+) and HD patients with anti-HCV(-)HBsAg(-). The durations of HD, hepatitis and liver cirrhosis were not related to the leukopenia or thrombocytopenia (p > 0.05). CONCLUSIONS: HCV infection associated with leukopenia and/or thrombocytopenia in HD patients is as common as in non-HD patients with liver cirrhosis. This may be due to the direct effect of hemopoiesis rather than the hyperspleenism of liver cirrhosis patients. There is a need for further prospective investigation to ascertain the clinical significance of leukopenia and thrombocytopenia in HD patients with anti-HCV(+). The prevalence of leukopenia and thrombocytopenia was higher in HD patients with hepatitis C than in HD patients with hepatitis B and HD patient without hepatitis.


Assuntos
Hepatite B/complicações , Hepatite C/complicações , Leucopenia/complicações , Cirrose Hepática/complicações , Diálise Renal , Trombocitopenia/complicações , Adulto , Idoso , Alanina Transaminase/sangue , Aspartato Aminotransferases/sangue , Biomarcadores/sangue , Feminino , Hepatite B/sangue , Antígenos de Superfície da Hepatite B/sangue , Hepatite C/sangue , Anticorpos Anti-Hepatite C/sangue , Humanos , Falência Renal Crônica/sangue , Falência Renal Crônica/complicações , Falência Renal Crônica/terapia , Contagem de Leucócitos , Cirrose Hepática/sangue , Masculino , Pessoa de Meia-Idade , Contagem de Plaquetas , Taiwan , Trombocitopenia/sangue
20.
Perit Dial Int ; 18(5): 516-21, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9848631

RESUMO

OBJECTIVE: To investigate the prevalence of nodular goiter and thyroid dysfunction in uremic patients undergoing hemodialysis (HD) and peritoneal dialysis. DESIGN: Cross-sectional study. SETTING: Single dialysis unit and outpatient clinic. PATIENTS: The study included 221 patients [143 HD and 78 continuous ambulatory peritoneal dialysis (CAPD) patients] along with 135 consecutively selected outpatients as controls. MAIN OUTCOME MEASURES: Ultrasonography was used to detect patients' thyroid function and nodular goiter. RESULTS: Nodular goiter was detected in 54.8% of the uremic patients and in 21.5% of the controls. Uremic patients had higher prevalence of thyroid dysfunction, which included reduced serum concentration of total T3, total T4, and free T4, and increased serum level of TSH. Hypothyroidism was also observed more frequently in uremic patients than in the control group (5.4% vs 0.7%, p < 0.05). Nodular goiter was more frequently found in females than in males (63.5% vs 48%, p < 0.05). Moreover, the prevalence of nodular goiter increased with age (p < 0.02) in uremic patients. Hemodialysis patients had a higher frequency of reduced total T3 level (46.9% vs 29.5%, p < 0.02). However, CAPD patients had lower T4 levels (6.23+/-1.82 microg/dL vs 7.15+/-1.99 microg/dL, p < 0.05). CONCLUSION: Because of the high incidence of hypothyroidism and nodular goiter in uremic patients, screening of thyroid function and goiter detection with ultrasound should be considered in evaluation of end-stage renal disease patients.


Assuntos
Bócio Nodular/epidemiologia , Hipertireoidismo/epidemiologia , Hipotireoidismo/epidemiologia , Diálise Peritoneal Ambulatorial Contínua , Diálise Renal , Estudos de Casos e Controles , Estudos Transversais , Feminino , Humanos , Incidência , Falência Renal Crônica/epidemiologia , Falência Renal Crônica/terapia , Masculino , Pessoa de Meia-Idade , Prevalência , Hormônios Tireóideos/sangue , Uremia/epidemiologia , Uremia/terapia
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