Expression of cytokine and chemokine mRNA and secretion of tumor necrosis factor-α by gallbladder epithelial cells: Response to bacterial lipopolysaccharides

BACKGROUND: In addition to immune cells, many other cell types are known to produce cytokines. Cultured normal mouse gallbladder epithelial cells, used as a model system for gallbladder epithelium, were examined for their ability to express the mRNA of various cytokines and chemokines in response to bacterial lipopolysaccharide. The synthesis and secretion of the tumor necrosis factor-α (TNF-α) protein by these cells was also measured. RESULTS: Untreated mouse gallbladder cells expressed mRNA for TNF-α, RANTES, and macrophage inflammatory protein-2 (MIP-2). Upon treatment with lipopolysaccharide, these cells now produced mRNA for Interleukin-1β (IL-1β), IL-6, monocyte chemoattractant protein-1 (MCP-1), and showed increased expression of TNF-α and MIP-2 mRNA. Untreated mouse gallbladder cells did not synthesize TNF-α protein; however, they did synthesize and secrete TNF-α upon treatment with lipopolysaccharide. METHODS: Cells were treated with lipopolysaccharides from 3 strains of bacteria. Qualitative and semi-quantitative RT-PCR, using cytokine or chemokine-specific primers, was used to measure mRNA levels of TNFα, IL-1β, IL-6, IL-10, KC, RANTES, MCP-1, and MIP-2. TNF-α protein was measured by immunoassays. CONCLUSION: This research demonstrates that gallbladder epithelial cells in response to lipopolysaccharide exposure can alter their cytokine and chemokine RNA expression pattern and can synthesize and secrete TNFα protein. This suggests a mechanism whereby gallbladder epithelial cells in vivo may mediate gallbladder secretory function, inflammation and diseases in an autocrine/paracrine fashion by producing and secreting cytokines and/or chemokines during sepsis

Laparoscopic versus open splenectomy in the management of hematologic diseases Esplenectomia laparoscópica versus aberta no tratamento de doenças hematológicas

Splenectomy is the best available treatment for severe forms of hereditary spherocytosis, idiopathic thrombocytopenic purpura, and other hematologic conditions when these prove refractory to conservative management. It has been employed for many decades with low mortality and favorable remission rates. The use of laparoscopic splenectomy in recent years has been rapidly and even enthusiastically adopted in this field. However, the exact role of laparoscopic versus open surgery for hematologic diseases is still debated. In this study of 58 adult patients, laparoscopic procedures were compared with conventional splenectomies for similar indications. METHODS: All patients were operated on within an 8-year period. Subjects underwent similar procedures under the supervision of the same surgical school and were compared regarding age, gender, body mass index, and diagnosis. Laparoscopically managed cases (Group I, n = 30) were prospectively followed according to a written protocol, whereas the same investigation was retrospectively done with regard to traditional laparotomy (Group II, n = 28). Methods included general and demographic findings, duration and technical steps of operation, blood loss, weight of spleen, need for conversion (in minimally invasive subjects), intraoperative and postoperative complications, time until realimentation, postoperative hospitalization, mortality, and late follow-up including recurrence rate. RESULTS: Idiopathic thrombocytopenic purpura was the surgical indication in over 50% of the patients in both groups, but familial spherocytosis, thalassemia, myelodysplasia, and lymphomas were also represented in this series. Laparoscopic procedures took more time to perform (P = 0.004), and postoperative hospitalization was 2 days shorter, but this difference was not statistically significant. Postoperative hematocrit and volume of blood transfusions was equivalent, although the laparoscopic cases had a somewhat lower preoperative hematocrit (NS) and displayed better recovery for this measurement (P = 0.03). More patients in Group I were able to accept oral food on the first day than subjects undergoing conventional operations (P < 0.05). Relatively few conversions were necessary during the minimally invasive surgeries (13.3%), and postoperative early and late complications as well as recurrences occurred in similar proportions. Also, the mean weight of the spleen was not statistically different between the groups, although there was a marked numerical tendency toward larger masses in conventional procedures. No spleen in Group I exceeded 2.0 kg, whereas in Group II values up to 4.0 kg occurred, and the mean weight was 50% higher in the latter group. CONCLUSIONS: 1) Minimally invasive splenectomy was essentially comparable to open surgery with regard to safety, efficacy, and late results; 2) Advantages concerning shorter postoperative hospitalization could not be shown, despite earlier food intake and a non-significant tendency toward earlier discharge; 3) This new modality should be considered an option in cases of hematologic conditions whenever the spleen is not hugely enlarged.<br>A esplenectomia é o melhor tratamento disponível para formas graves de esferocitose hereditária, púrpura trombocitopênica idiopática e outras entidades hematológicas refratárias à abordagem conservadora. Ela tem sido empregada há muitas décadas com baixa mortalidade e taxas de remissão favoráveis. A alternativa de esplenectomia laparoscópica em anos recentes foi adotada rapidamente e até entusiasticamente, todavia o papel exato das intervenções abertas em contraposição às laparoscópicas para doenças hematológicas ainda é objeto de debate. Num estudo de 58 pacientes adultos, os procedimentos laparoscópicos foram comparados com as esplenectomias convencionais em indicações semelhantes. MÉTODOS: Todos os pacientes foram operados num período de 8 anos. Eles foram submetidos a operações análogas sob a supervisão da mesma escola cirúrgica e eram comparáveis no tocante a idade, sexo, índice de massa corporal e diagnóstico. Os casos abordados laparoscopicamente (Grupo I, n= 30) foram seguidos prospectivamente de acordo com protocolo escrito, ao passo que a mesma investigação foi aplicada retrospectivamente no que concerne aos doentes de esplenectomia aberta (Grupo II, n= 28). Os métodos incluíram achados gerais e demográficos, duração e passos técnicos da cirurgia, perda sangüínea, peso do baço, necessidade de conversão (nos casos minimamente invasivos), complicações intra e pós-operatórias, tempo para realimentação, hospitalização pós-operatória, mortalidade e seguimento tardio, incluindo recidivas. RESULTADOS: A púrpura trombocitopênica idiopática foi a indicação cirúrgica em mais de 50% dos enfermos de ambos os grupos, entretanto esferocitose familiar, talassemia, mielodisplasia e linfomas também estavam representados nesta série. As intervenções laparoscópicas demoraram mais (p=0,004) e sua hospitalização pós-operatória foi dois dias mais breve, porém esta diferença não foi estatisticamente significativa. O hematócrito pós-operatório e o volume de transfusões foram equivalentes, no entanto os casos laparoscópicos exibiam um hematócrito pré-operatório ligeiramente inferior (NS) e a recuperação desta variável foi melhor (p=0,03). Mais pacientes do Grupo I toleraram dieta oral no primeiro dia que casos abordados convencionalmente (p<0,05). Relativamente poucas conversões foram necessárias no decurso das laparoscopias (13,3%), e as complicações pós-operatórias precoces e tardias assim como as recidivas distribuíram-se de forma eqüitativa. Também não foi possível demonstrar-se diferenças no peso do baço, ainda que no Grupo I nenhum órgão excedesse a 2,0 kg, sendo que no Grupo II este valor chegou até 4,0 kg, notando-se ainda que o peso médio foi 50% mais elevado nesta última população. CONCLUSÕES: 1) A esplenectomia minimamente invasiva foi essencialmente comparável à variante aberta no tocante à segurança, eficácia e resultados tardios; 2) Não foi possível comprovar-se vantagens relativas à menor hospitalização pós-operatória, embora a realimentação fosse mais precoce e houvesse uma tendência não-significativa para alta hospitalar antecipada; 3) Esta nova modalidade deve ser considerada uma excelente opção para casos de moléstias hematológicas sempre que o baço não estiver fortemente aumentado