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1.
Am J Transplant ; 15(6): 1708-15, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25754742

RESUMO

Improved understanding of risk factors associated with carbapenem-resistant-Klebsiella pneumoniae (CR-KP) infection after liver transplantation (LT) can aid development of effective preventive strategies. We performed a prospective cohort study of all adult patients undergoing LT at our hospital during 30-month period to define risk factors associated with CR-KP infection. All patients were screened for CR-KP carriage by rectal swabs before and after LT. No therapy was administered to decolonize or treat asymptomatic CR-KP carriers. All patients were monitored up to 180 days after LT. Of 237 transplant patients screened, 41 were identified as CR-KP carriers (11 at LT, 30 after LT), and 20 developed CR-KP infection (18 bloodstream-infection, 2 pneumonia) a median of 41.5 days after LT. CR-KP infection rates among patients non-colonized, colonized at LT, and colonized after LT were 2%, 18.2% and 46.7% (p < 0.001). Independent risk factors for CR-KP infection identified by multivariate analysis, included: renal-replacement-therapy; mechanical ventilation > 48 h; HCV recurrence, and colonization at any time with CR-KP. Based on these four variables, we developed a risk score that effectively discriminated patients at low versus higher risk for CR-KP infection (AUC 0.93, 95% CI 0.86-1.00, p < 0.001). Our results may help to design preventive strategies for LT recipients in CR-KP endemic areas.


Assuntos
Carbapenêmicos/uso terapêutico , Portador Sadio/microbiologia , Farmacorresistência Bacteriana , Infecções por Klebsiella/tratamento farmacológico , Infecções por Klebsiella/epidemiologia , Klebsiella pneumoniae/isolamento & purificação , Transplante de Fígado , Adulto , Idoso , Estudos de Coortes , Contagem de Colônia Microbiana , Feminino , Humanos , Incidência , Infecções por Klebsiella/diagnóstico , Masculino , Programas de Rastreamento/métodos , Pessoa de Meia-Idade , Modelos Estatísticos , Análise Multivariada , Estudos Prospectivos , Fatores de Risco , Fatores de Tempo
2.
Case Rep Transplant ; 2014: 262953, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25177510

RESUMO

An adult male underwent a bowel transplant for tufting enteropathy, receiving alemtuzumab, tacrolimus, and steroids as immunosuppressants. Five years later, he developed an autoimmune hemolytic anemia (AIHA), anti-IgG positive, with reduced reticulocyte count, leukopenia, and thrombocytopenia with antiplatelet antibodies. After an unsuccessful initial treatment with high dose steroids, reduction in tacrolimus dose, and intravenous immunoglobulin (IVIG), a bone marrow biopsy revealed absence of erythroid maturation with precursor hyperplasia. The patient was switched to sirolimus and received four doses of rituximab plus two courses of plasmapheresis, which decreased his transfusion requirements. After a febrile episode one month later, the AIHA relapsed with corresponding decreases in platelet and leukocyte count: cyclosporine A (CsA) was started with a second course of rituximab and IVIG without response, even though repeat bone marrow biopsy did not reveal morphology correlated to an acquired pure red cell aplasia (APRCA). Considering the similarity in his clinical and laboratory findings to APRCA, alemtuzumab was added (three doses over a week) with CsA followed by steroids. The patient was eventually discharged transfusion-independent, with increasing hemoglobin (Hb) levels and normal platelet and leukocyte count. One year later he is still disease-free with functioning graft.

3.
J Econ Entomol ; 103(3): 816-22, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20568628

RESUMO

The pear psylla, Cacopsylla pyri L. (Hemiptera: Psyllidae), is a relevant pest of pear, Pyrus communis L., trees in Emilia-Romagna region (northern Italy). The susceptibility to the insecticide abamectin was evaluated at different times of the year on C. pyri populations undergoing different control strategies within conventional, integrated, and organic farms. The tests performed were the egg spray and the topic and dip bioassay on adults. The larval mortality was evaluated by dip bioassay on treated leaves. The activity of P450-dependent monooxygenases, a relevant enzyme system involved in insecticide resistance of C. pyri, was also determined in adults by 7-ethoxycoumarin O-deethylation (ECOD assay). Tests on treated eggs and on larvae showed no significant differences in LC50 and LC90, although these values were always lower in individuals collected from organic farms in comparison with all other farms. Tests on overwintering adults revealed differences among populations, probably more related to collection time than to field pest control strategies. Unexpectedly, the ECOD assay on adults showed a slightly higher cytochrome P450 monooxygenase activity in the population undergoing organic control in comparison to others. Our results indicate that egg spray is the most reliable bioassay to verify data of open-field applications. Apparently, no resistance to abamectin has yet been developed by C. pyri in Emilia-Romagna.


Assuntos
Hemípteros , Inseticidas , Ivermectina/análogos & derivados , Animais , Sistema Enzimático do Citocromo P-450/metabolismo , Hemípteros/enzimologia , Itália , Larva , Dose Letal Mediana , Óvulo , Pyrus
4.
J Hazard Mater ; 171(1-3): 524-34, 2009 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-19619943

RESUMO

Sites contaminated with hazardous material are a topical and urgent problem all over the world. In accordance with recent Italian regulations, appropriate risk assessment is required in order to determine health risks associated with contaminated sites. The paper presents a case study regarding a disused industrial plant contaminated with polycyclic aromatic hydrocarbons, heavy hydrocarbons and polychlorinated biphenyls. The site is characterized by three different topographical levels. Therefore both the characterization and the conceptual model had to be adapted to the site conditions: we divided the site into three discrete areas and we developed a separate risk assessment for each area. Besides health risk assessment, we performed ecological risk assessment for both groundwater and surface water targets, as required by Italian regulations. The future reuse scenario has not yet been defined and, consequently, risk assessment results will be useful for the remediation program. Risk assessment was supported by leaching tests and hydrocarbon "finger printing". Leaching tests allowed us to determine site-specific soil-water partition coefficient. Hydrocarbon "finger printing" allowed us to differentiate the mobility of the different hydrocarbon groups in migration analyses. We found the site required remediation based on Italian standard. We propose a simple risk-based remediation action consisting in the replacement of the upper 1m with "clean" soil and the placement of a barrier to vapors.


Assuntos
Resíduos Industriais , Monitoramento Ambiental/métodos , Geografia , Humanos , Hidrocarbonetos/toxicidade , Itália , Bifenilos Policlorados/análise , Risco , Medição de Risco , Solo , Poluentes do Solo/análise , Poluentes do Solo/toxicidade , Movimentos da Água , Poluentes Químicos da Água/toxicidade , Abastecimento de Água
5.
Minerva Chir ; 58(3): 341-4, 2003 Jun.
Artigo em Italiano | MEDLINE | ID: mdl-12955053

RESUMO

BACKGROUND: The availability of drugs which can effectively inhibit gastric secretion allowed the control of the peptic disease in almost all the patients, but the incidence of complications is substantially unchanged. METHODS: In the present study, we performed a retrospective evaluation of 153 patients treated for complicated peptic ulcer (hemorrhage and perforation) in the last 6 years. Hemor-rhage is a common onset of the peptic disease and it rarely requires a surgical treatment, because medical and endoscopic therapies are successful in a high number of patients. RESULTS: Surgery was necessary in 17.2% of the cases in the group of patients studied. CONCLUSIONS: Our experience confirms the high operative mortality in the patients who underwent surgery for hemorrhage related to complicated peptic ulcer because of the frequent presence of several risk factors. Surgical therapy is the main procedure of treatment of perforated peptic ulcer and in the group of patient examined all the cases of perforation underwent surgical operation. In conclusion, despite the progress of pharmacological and endoscopic therapies, surgical treatment is the best therapy of complicated peptic ulcer.


Assuntos
Úlcera Péptica/complicações , Úlcera Péptica/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos
6.
Bull Entomol Res ; 93(4): 335-42, 2003 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-12908919

RESUMO

Peach shoot volatiles were attractive to mated female oriental fruit moth, Cydia molesta (Busck), in a dual choice arena. No preference was observed between leaf odours from the principle host plant, peach, and the secondary host plant, apple. Twenty-two compounds were identified in headspace volatiles of peach shoots using gas chromatography-mass spectrometry. Green leaf volatiles accounted for more than 50% of the total emitted volatiles. A bioassay-assisted fractionation using different sorbent polymers indicated an attractant effect of compounds with a chain length of 6-8 carbon atoms. The major compounds of this fraction were tested either singly or in combinations for behavioural response of females. Significant bioactivity was found for a three-component mixture of (Z)-3-hexen-1-yl acetate, (Z)-3-hexen-1-ol and benzaldehyde in a 4:1:1 ratio. This synthetic mixture elicited a similar attractant effect as the full natural blend from peach shoots as well as the bioactive fraction.


Assuntos
Lepidópteros/fisiologia , Malus/química , Odorantes/análise , Prunus/química , Animais , Bioensaio/veterinária , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Malus/parasitologia , Brotos de Planta/química , Prunus/parasitologia , Volatilização
7.
Ann Ital Chir ; 73(2): 173-7; discussion 178-80, 2002.
Artigo em Italiano | MEDLINE | ID: mdl-12197291

RESUMO

AIM: In elderly people colorectal cancer has an high incidence of emergency presentation. In this article we report our experience on colorectal emergency surgery for cancers in patients older than 80 yrs. METHODS: From October 1995 to December 2000 323 patients were submitted to emergency surgical procedure for colorectal cancer at the Dpt of Emergency Surgery of University of Bologna. 37 (11%) subjects were over 80 years of age. A retrospective analysis was performed: the data collected included the mode of presentation of the patients, the location and Dukes' classification of the tumors, the incidence of radical and palliative operations, the causes of death and 30-day mortality. RESULTS: The most common emergency presentation was an intestinal occlusion (21 cases); the other different presentations were peritonitis (7 cases) and hemorrhage (9 cases). 22 patients underwent resection of the primary growth and anastomosis and 11 subjects had palliative intervention by creating a stoma only or bypass anastomosis without resection, 2 patients had an explorative laparotomy and 2 were inoperable. Our overall postoperative mortality was 14.2% and we registered 1 anastomotic leakage. DISCUSSION AND CONCLUSIONS: Emergency surgery for colorectal cancer in patients over 80 years of age can be performed safely without restrictions related to the age.


Assuntos
Idoso , Neoplasias Colorretais/cirurgia , Fatores Etários , Idoso de 80 Anos ou mais , Causas de Morte , Neoplasias Colorretais/complicações , Neoplasias Colorretais/mortalidade , Emergências , Feminino , Humanos , Masculino , Cuidados Paliativos , Estudos Retrospectivos , Fatores Sexuais , Fatores de Tempo
8.
Minerva Chir ; 57(3): 363-9, 2002 Jun.
Artigo em Inglês, Italiano | MEDLINE | ID: mdl-12029232

RESUMO

BACKGROUND: Incisional hernia (IH) repair with conventional techniques is associated with high recurrence rate. Surgical repair using prosthetic biomaterials is becoming increasingly popular. On the basis of the good results an increasing number of surgeons have begun to use this technique. However prosthesis use in contaminated fields is still debated. In complicated IH contaminating surgical procedure are often performed and the use of meshes can be hazardous. The aim of this study was to report our experience about the treatment of complicated IH with prosthetic materials in an emergency surgery setting. METHODS: From November 1995 to November 2001 55 patients were submitted to emergency surgical treatment for complicated IH. Eleven patients were treated using a prosthetic device. Patients mean age was 71.3 (range 38-91). About 70% of patients had concomitant major cardiac or pulmonary diseases. RESULTS: Thirty-two IH were strangulated, 23 were incarcerated. In all patients a viscerolysis was carried out. In 6 patients a small bowel resection was done and in 4 subjects a large bowel resection was performed. Eight patients were submitted to omental resection. There were not differences in morbidity and mortality between the studied group. Recurrence rate was significantly lower in prosthetic treated group. CONCLUSIONS: We concluded that prosthetic repair of complicated IH is feasible in selected cases allowing abdominal wall anatomy re-establishment.


Assuntos
Tratamento de Emergência , Hérnia Ventral/complicações , Hérnia Ventral/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Perfuração Intestinal/etiologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Telas Cirúrgicas , Resultado do Tratamento
9.
EMBO J ; 20(21): 5950-61, 2001 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-11689435

RESUMO

To obtain a better understanding of how hippocampal neurons selectively target proteins to axons, we assessed whether any of the large cytoplasmic regions of neuronal sodium channel Na(v)1.2 contain sufficient information for axonal compartmentalization. We show that addition of the cytoplasmic C-terminal region of Na(v)1.2 restricted the distribution of a dendritic-axonal reporter protein to axons. The analysis of mutants revealed that a critical segment of nine amino acids encompassing a di-leucine-based motif mediates axonal compartmentalization of chimera. In addition, the Na(v)1.2 C-terminus is recognized by the clathrin endocytic pathway both in non-neuronal cells and the somatodendritic domain of hippocampal neurons. The mutation of the di-leucine motif located within the nine amino acid sequence to alanines resulted in the loss of chimera compartmentalization in axons and of internalization. These data suggest that selective elimination by endocytosis in dendrites may account for the compartmentalized distribution of some proteins in axons.


Assuntos
Axônios/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Neurônios/metabolismo , Canais de Sódio/metabolismo , Motivos de Aminoácidos/fisiologia , Sequência de Aminoácidos , Animais , Células COS , Compartimento Celular/fisiologia , Células Cultivadas , Dendritos/metabolismo , Genes Reporter , Células HeLa , Hipocampo/citologia , Hipocampo/metabolismo , Humanos , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Canal de Sódio Disparado por Voltagem NAV1.2 , Proteínas do Tecido Nervoso/genética , Neurônios/citologia , Transporte Proteico/fisiologia , Ratos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Canais de Sódio/genética , Relação Estrutura-Atividade
10.
Dig Surg ; 17(5): 503-7, 2000.
Artigo em Inglês | MEDLINE | ID: mdl-11124556

RESUMO

BACKGROUND/AIMS: Gastrointestinal stromal tumors (GISTs) are a rare group of neoplasias of the gastrointestinal tract that have not yet been fully investigated. In this article the authors present the experience of an emergency surgery department in the diagnosis and treatment of patients with such neoplasms and discuss the approaches to these 'strange' tumors. METHODS: A review of our 4-year experience in emergency surgery was performed and 9 patients were found with the diagnosis of GIST. The median follow-up was 32.3 (range 18-45) months. RESULTS: 7 patients had evidence of gastrointestinal blood loss, 1 patient had abdominal pain, and the last patient had anorexia, vomiting and fever. Five tumors were located in the stomach and 4 in the small bowel. All the patients underwent complete resection. On histological examination 5 tumors were of myogenic origin, 1 was a gastrointestinal autonomic nerve tumor and 1 was a mixed neural-myoid tumor. The remaining 2 could not be differentiated. Of the 9 patients who underwent curative resections, 1 had a recurrence and died. CONCLUSION: GIST treatment mainly involves surgical resection with the goal of complete removal which can be curative. The histologic grade and tumor size are the most important prognostic factors.


Assuntos
Neoplasias do Íleo/cirurgia , Neoplasias do Jejuno/cirurgia , Neoplasias Gástricas/cirurgia , Adulto , Idoso , Serviço Hospitalar de Emergência , Feminino , Humanos , Neoplasias do Íleo/diagnóstico , Neoplasias do Jejuno/diagnóstico , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Retrospectivos , Neoplasias Gástricas/diagnóstico
11.
J Cell Biochem ; 79(4): 628-47, 2000 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-10996854

RESUMO

Bile salt-dependent lipase (BSDL) was detected in human SOJ-6 and rat AR4-2J pancreatic cells. Whereas AR4-2J cells actively secreted the enzyme, BSDL was retained within the Golgi compartment of SOJ-6 cells. Because Rab6 is involved in vesicle transport in the Golgi apparatus and the trans-Golgi network, we confirmed the presence of Rab6 in these cells. In rat AR4-2J cells, Rab6 as well as Rab1A/B and Rab2, partitioned between the cytosol and microsomes. In SOJ-6 cells Rab1A/B and Rab2 also partitioned between the cytosol and microsomes, but Rab6 was strictly associated with microsome membranes, suggesting a specific defect of Rab6 cycling in human SOJ-6 cells. The apparent defect of cycling in these cells is not due to the expression of a defective Rab6 since its correct sequence was confirmed. We further demonstrated that AR4-2J and SOJ-6 cells express the Rab-GDIbeta and Rab-GDIalpha isoforms, respectively. However, the sequence of Rab-GDIbeta, which may be the main form expressed by SOJ-6 cells, identified a few substitutions located in regions that are essential for Rab-GDI function. We conclude that the deficient secretion of BSDL by SOJ-6 cells could be due to the expression of defective Rab-GDIbeta. In spite of the alterations in Rab-GDIbeta, membrane proteins such as CD71 and NHE3 were correctly localized to the cell plasma membrane of SOJ-6 cells, suggesting that two functional distinct secretory pathway coexist in pancreatic cells.


Assuntos
Neoplasias Pancreáticas/enzimologia , Esterol Esterase/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Ácidos e Sais Biliares/metabolismo , Transporte Biológico Ativo , Primers do DNA/genética , Inibidores de Dissociação do Nucleotídeo Guanina/genética , Inibidores de Dissociação do Nucleotídeo Guanina/metabolismo , Humanos , Glicoproteínas de Membrana/metabolismo , Dados de Sequência Molecular , Mutação , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Neoplásico/genética , RNA Neoplásico/metabolismo , Ratos , Homologia de Sequência de Aminoácidos , Esterol Esterase/genética , Células Tumorais Cultivadas , Proteínas rab de Ligação ao GTP/genética , Proteínas rab de Ligação ao GTP/metabolismo
12.
Arch Biochem Biophys ; 375(1): 90-100, 2000 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-10683253

RESUMO

This work describes the characterization of an immunoreactive form of bile salt-dependent lipase (BSDL) expressed by the human pancreatic tumoral Mia PaCa-2 cell line. This BSDL-related protein, which has an M(r) of 70 kDa, is enzymatically active and poorly secreted. Furthermore, a protein with the same electrophoretic migration can also be immunoprecipitated with polyclonal antibodies specific for the human pancreatic BSDL after in vitro translation of RNA isolated from Mia PaCa-2 cells. These data indicated that this BSDL-related protein might be poorly, or not, glycosylated. Reverse transcription and amplification of RNA extracted from Mia PaCa-2 cells using primers able to specifically amplify the full-length mRNA of the human BSDL resulted in a detectable 1.8-kb cDNA product, shorter than that of BSDL (2.2 kb). The sequence of this transcript corresponds to the mRNA sequence that codes for the mature human pancreatic BSDL. However, a deletion of 330 bp is located within the 3'-domain of this cDNA. Therefore data allowed us to conclude that the 70-kDa BSDL-related protein is a 612 amino acid length protein and represents a truncated form of BSDL. The deletion of 110 amino acids occurs in the C-terminal region of the protein, which encompasses 6 tandemly repeated sequences instead of the 16 normally present in the sequence of BSDL. Because feto-acinar pancreatic protein (FAPP), which is the oncofetal counterpart of BSDL, is a C-terminally truncated isoform of BSDL, it is suggested that the 70-kDa BSDL-related protein expressed in MiaPaCa-2 cells could be representative of the protein moiety of FAPP.


Assuntos
Lipase , Neoplasias Pancreáticas/enzimologia , Esterol Esterase/biossíntese , Esterol Esterase/imunologia , Processamento Alternativo , Sequência de Aminoácidos , Sequência de Bases , Proteínas de Transporte/genética , Sistema Livre de Células/metabolismo , Glicoproteínas/genética , Glicosilação , Humanos , Imunoadsorventes , Isoenzimas/biossíntese , Isoenzimas/genética , Isoenzimas/imunologia , Isoenzimas/isolamento & purificação , Dados de Sequência Molecular , Pâncreas/metabolismo , Neoplasias Pancreáticas/patologia , Biossíntese de Proteínas , RNA Mensageiro/biossíntese , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Esterol Esterase/genética , Esterol Esterase/isolamento & purificação , Sequências de Repetição em Tandem/genética , Células Tumorais Cultivadas
13.
Biochem J ; 345 Pt 1: 121-8, 2000 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-10600647

RESUMO

Bile-salt-dependent lipase (BSDL, EC 3.1.1.-) is an enzyme expressed by the pancreatic acinar cells and secreted as a component of the pancreatic juice of all examined species. During its secretion route BSDL is associated with intracellular membranes. This association allows the complete glycosylation of the enzyme or participates in the inhibition of the enzyme activity, which can deleterious for the acinar pancreatic cell. Thereafter, the human BSDL is phosphorylated by a serine/threonine protein kinase and released from intracellular membranes. In the present study, we show that the rat pancreatic BSDL, expressed by AR4-2J cells used as a model, is phosphorylated by a protein kinase that is insensitive to inhibitors of protein kinases A, C or G and that the phosphorylation process is favoured by okadaic acid (an inhibitor of protein phosphatases 1 and 2A). However, 5,6-dichloro-1-beta-d-ribofuranosylbenzimidazole (DRB), which is a specific inhibitor of casein kinase II, abolishes the phosphorylation in vitro of BSDL within micro- somes of AR4-2J pancreatic cells. We showed further that the alpha-subunit of casein kinase II co-locates with BSDL within the lumenal compartment of the Golgi. Genistein, which perturbs the trans-Golgi network, also inhibits the phosphorylation of BSDL, suggesting that this post-translational modification of BSDL probably occurred within this cell compartment. The inhibition of the phosphorylation of BSDL by DRB also decreases the rate at which the enzyme is secreted. Under the same conditions, the rate of alpha-amylase secretion was not modified. These data strongly suggest that phosphorylation is a post-translational event, which appears to be essential for the secretion of BSDL.


Assuntos
Pâncreas/enzimologia , Proteínas Serina-Treonina Quinases/metabolismo , Esterol Esterase/metabolismo , Animais , Caseína Quinase II , Compartimento Celular , Linhagem Celular , Diclororribofuranosilbenzimidazol/farmacologia , Inibidores Enzimáticos/farmacologia , Genisteína/farmacologia , Humanos , Técnicas In Vitro , Fosforilação , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/antagonistas & inibidores , Ratos , Esterol Esterase/química , alfa-Amilases/metabolismo
14.
Glycobiology ; 9(9): 935-46, 1999 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-10460835

RESUMO

The feto-acinar pancreatic protein or FAPP, the oncofetal glycoisoform of bile salt-dependent lipase (BSDL), is characterized by the presence of the J28 glycotope recognized by mAbJ28. This fucosylated epitope is carried out by the O-linked glycans of the C-terminal mucin-like region of BSDL. This glycotope is expressed by human tumoral pancreatic tissues and by human pancreatic tumoral cell lines such as SOJ-6 and BxPC-3 cells. However, it is not expressed by the normal human pancreatic tissues and by MiaPaCa-2 and Panc-1 cells. Due to the presence of many putative sites for O-glycosylation on FAPP and BSDL, the structure of the J28 glycotope cannot be attained by classical physical methods. In the first part of the present study, we have determined which glycosyltransferases were differently expressed in pancreatic tumoral cell lines compared to normal tissues, focusing in part on fucosyltransferases (Fuc-T) and core-2 beta6-N-acetylglucosaminyltransferase (Core2GlcNAc-T). Our data suggested that alpha2-Fuc-T activity was decreased in the four cell lines tested (SOJ-6, BxPC-3, MiaPaCa-2, and Panc-1). The alpha(1-3) and alpha(1-4) fucosylations were decreased in tumor cells that do not express the J28 glycotope whereas alpha4-Fuc-T and Core2GlcNAc-T activities were significantly increased in SOJ-6 cells which best expressed the J28 glycotope. Therefore, we wished to gain information about glycosyltransferases involved in the building of this structure by transfecting the cDNA encoding the mucin-like region of BSDL in CHO-K1 also expressing Core2GlcNAc-T and/or FUT3 and/or FUT7 activities. These CHO-K1 cells have been previously transfected with the cDNA encoding Core2GlcNAc-T and/or FUT3 and/or FUT7. Data indicated that the C-terminal peptide of BSDL (Cter) produced by those cells did not carry out the J28 glycotope unless Core2GlcNAc-T activity is present. Further transfection with FUT3 cDNA, increased the antibody recognition. Nevertheless, transfection with FUT3 or FUT7 alone did not generate the formation of the J28 glycotope on the C-terminal peptide. Furthermore, the Cter peptide produced by CHO-K1 cells expressing Core2GlcNAc-T was more reactive to the mAbJ28 after in vitro fucosylation with the recombinant soluble form of FUT3. These data suggested that the J28 glycotope encompasses structures initiated by Core2GlcNAc-T and further fucosylated by alpha3/4-Fuc-T such as FUT3, likely on GlcNAc residues.


Assuntos
Antígenos de Neoplasias/biossíntese , Epitopos/biossíntese , Fucosiltransferases/metabolismo , Glicoproteínas/biossíntese , N-Acetilglucosaminiltransferases/metabolismo , Esterol Esterase/biossíntese , Clonagem Molecular , DNA Complementar/genética , Glicoproteínas/imunologia , Glicosilação , Humanos , Neoplasias Pancreáticas/química , Neoplasias Pancreáticas/enzimologia , Fragmentos de Peptídeos/metabolismo , Esterol Esterase/genética , Esterol Esterase/imunologia , Células Tumorais Cultivadas
15.
Biochem J ; 342 ( Pt 1): 179-87, 1999 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-10432315

RESUMO

In this article, we report the nucleotide sequence of the cDNA encoding an isoform of bile-salt-dependent lipase (BSDL) expressed by human hepatoma cells. The BSDL is a 100-kDa glycoprotein normally expressed by the human pancreas. Using a polyclonal antibody raised against an internal peptide located between Ile(327) and Glu(350) of the human pancreatic BSDL, we have immunodetected an isoform of human pancreatic BSDL, with an apparent molecular mass of about 62 kDa. This isoform of BSDL was mainly associated with the cytosol of a human hepatoma cell line (HepG2), the remaining protein being found in the microsome fraction. In addition, esterolytic activity on p-nitrophenyl hexanoate measured in microsomes and cytosol appeared very low and was weakly stimulated by bile salts, such as taurocholate. In contrast to human pancreatic BSDL, which is secreted as a component of pancreatic juice, this isoform appeared to be retained in the HepG2 cells. Reverse transcription, followed by PCR and amplification, performed on RNA extracted from HepG2 cells using specific primers hybridizing to the sequence coding for the entire normal human pancreatic BSDL, allowed us to amplify a 1. 7-kb transcript that appeared to be 0.5 kb shorter than the transcript of the pancreatic enzyme (2.2 kb). From the sequence of the transcript thus obtained, a protein with a molecular mass of 62 kDa might be predicted, which is in good agreement with the size of the isoform of BSDL immunodetected in HepG2 cells. The N-terminal amino-acid sequence, deduced from the 1.7-kb transcript sequence, matched that of the pancreatic BSDL. However, the C-terminal domain appeared truncated, bearing only a single mucin-like sequence compared with sixteen for the human pancreatic BSDL. The actual intracellular function of this human BSDL hepatoma isoform remains to be elucidated.


Assuntos
Microssomos Hepáticos/enzimologia , Esterol Esterase/genética , Esterol Esterase/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Ácidos e Sais Biliares/farmacologia , Western Blotting , Clonagem Molecular , Reações Cruzadas , Citosol/efeitos dos fármacos , Citosol/enzimologia , Humanos , Isoenzimas/química , Isoenzimas/genética , Isoenzimas/imunologia , Isoenzimas/metabolismo , Neoplasias Hepáticas/enzimologia , Microssomos Hepáticos/efeitos dos fármacos , Dados de Sequência Molecular , Peso Molecular , Pâncreas/enzimologia , Biossíntese de Proteínas , RNA Mensageiro/análise , RNA Mensageiro/genética , Esterol Esterase/química , Esterol Esterase/imunologia , Células Tumorais Cultivadas
16.
Minerva Chir ; 53(9): 743-5, 1998 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-9866943

RESUMO

Meckel's diverticulum is the most common congenital abnormality of the intestinal tract, occurring in 2% of autoptic studies. The case of an 85-year-old man referred to the Emergency Surgery Unit for intestinal obstruction and lower gastrointestinal tract bleeding is reported. Surgical exploration revealed a complicated Meckel's diverticulum full of coproliths, immersed in pus and blood. Examination of the resected diverticulum showed necrotic diverticulitis in the absence of ectopic gastric or pancreatic tissues.


Assuntos
Divertículo Ileal/cirurgia , Doença Aguda , Idoso , Idoso de 80 Anos ou mais , Impacção Fecal/diagnóstico , Impacção Fecal/etiologia , Impacção Fecal/patologia , Impacção Fecal/cirurgia , Humanos , Masculino , Divertículo Ileal/complicações , Divertículo Ileal/diagnóstico , Divertículo Ileal/patologia
17.
Biochim Biophys Acta ; 1408(1): 44-54, 1998 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-9784601

RESUMO

The mechanisms by which ethanol administration alters pancreatic function are unknown. We have evaluated the effects of chronic ethanol treatment on secretion of a digestive enzyme: the bile salt-dependent lipase (BSDL), by the rat pancreatic cell line AR4-2J (as a model). We report that ethanol (50-300 mM) in culture medium induced a rise, in secreted and intracellular BSDL, that was a function of the duration of treatment and of the ethanol concentration. This effect was not abolished by pyrazole, which suggests a direct effect of ethanol. We have further established that the increase of BSDL activity was due to an enhanced biosynthesis of the enzyme consecutive to a major steady-state level of mRNA encoding BSDL. Also, the subcellular localization showed a specific accumulation of BSDL in the cytosolic fraction of cells chronically treated with ethanol. Given the enzymatic properties of BSDL, all these data could have some physiological consequences regarding the digestive function, plasma lipid metabolism and intracellular cholesterol homeostasis.


Assuntos
Etanol/farmacologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Esterol Esterase/genética , Animais , Relação Dose-Resposta a Droga , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Pâncreas/enzimologia , Neoplasias Pancreáticas , Pirazóis/farmacologia , RNA Mensageiro/metabolismo , Ratos , Esterol Esterase/biossíntese , Transcrição Gênica/efeitos dos fármacos , Células Tumorais Cultivadas
18.
J Biol Chem ; 273(43): 28208-18, 1998 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-9774442

RESUMO

Specific transcripts for bile salt-dependent lipase (BSDL), a 100-kDa glycoprotein secreted by the human pancreas, were immunodetected in BxPC-3 and SOJ-6 pancreatic tumoral cell lines. Sequencing of fragments, obtained by mRNA reverse transcription and amplification, confirmed the presence of BSDL transcripts in these cancer cells. The protein was detected in lysates of pancreatic tumoral cells, where it was mainly associated with membranes. Only a minute amount of the enzyme was detected in the culture media. Immunofluorescence studies demonstrated that in SOJ-6 cells, BSDL colocates with the p58 Golgi protein and suggested that the protein may be sequestrated within the Golgi compartment. These results demonstrated that BSDL is expressed in human pancreatic tumoral cells and cannot be secreted (or for the least very poorly). Subsequently, a cDNA covering the entire sequence of BSDL was obtained by reverse transcription-polymerase chain reaction. The sequence of this cDNA indicated that the N-terminal domain encoded by exons 1-10 was identical to that of BSDL expressed by the human normal pancreas. However, the sequence corresponding to exon 11, which should code for the 16 tandem-repeated identical mucin-like sequences of BSDL, was deleted by 330 base pairs (bp) and encoded only 6 of these repeated sequences. We conclude that this truncated variant of BSDL would be its oncofetal form, referred to as feto-acinar pancreatic protein. We then investigated whether the deletion of 330 bp affected the secretion of the protein. For this purpose, the cDNA corresponding to the mature form of the BSDL variant expressed in SOJ-6 cells was cloned into an expression/secretion vector and transfected into CHO-K1 cells. Results indicated that the variant of BSDL isolated from SOJ-6 cells was expressed and secreted by transfected cells. However, the level of BSDL secreted by these transfected CHO-K1 cells was significantly higher than that observed for SOJ-6 cells. Consequently, the retention of the oncofetal variant of BSDL observed in human pancreatic tumoral cells might not result from inherent properties of the protein.


Assuntos
Proteínas de Transporte/genética , Glicoproteínas/genética , Isoenzimas/genética , Lipase , Proteínas de Neoplasias/genética , Neoplasias Pancreáticas/enzimologia , Esterol Esterase/genética , Sequência de Aminoácidos , Aminoácidos/análise , Sequência de Bases , Compartimento Celular , Complexo de Golgi , Humanos , Proteínas de Membrana/genética , Dados de Sequência Molecular , Mutação , Fragmentos de Peptídeos/química , RNA Mensageiro/análise , Sequências Repetitivas de Ácido Nucleico , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Células Tumorais Cultivadas
19.
Minerva Chir ; 53(4): 285-7, 1998 Apr.
Artigo em Italiano | MEDLINE | ID: mdl-9701983

RESUMO

The gallbladder volvulus is a very rare but extremely dangerous event. Described for the first time by Wendel in 1898, it affects more frequently the female sex, particularly in old age. The clinical case of a patient affected by this pathology, personally observed, is presented and the initial symptoms, diagnostic procedures and surgical intervention are described in detail. The anatomical and physiological backgrounds that permit the gallbladder to twist on its axis are analysed and finally the importance of an early diagnosis to avoid the complications of a bilious peritonitis is underlined. In any case the definite diagnosis is made more frequently during surgical intervention. An early intervention allows a rapid resolution of the clinical picture preventing the perforation of the viscus into the peritoneal cavity and the complication of a bilious peritonitis besides the spreading of the biliary stones into the abdominal cavity.


Assuntos
Colecistectomia , Doenças da Vesícula Biliar/cirurgia , Idoso , Idoso de 80 Anos ou mais , Colelitíase/complicações , Feminino , Vesícula Biliar/irrigação sanguínea , Vesícula Biliar/patologia , Gangrena , Humanos , Isquemia/etiologia , Isquemia/cirurgia , Necrose , Peritonite/prevenção & controle , Anormalidade Torcional/complicações , Anormalidade Torcional/cirurgia
20.
Glycobiology ; 8(6): 605-13, 1998 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-9592127

RESUMO

Human pancreatic cancer is characterized by an alteration in fucose-containing surface blood group antigens such as H antigen, Lewis b, Lewis y, and sialyl-Lewis. These carbohydrate determinants can be synthesized by sequential action of alpha(2,3) sialyltransferases or alpha(1,2) fucosyltransferases (Fuc-T) and alpha(1,3/1,4) fucosyltransferases on (poly)N-acetyllactosamine chains. Therefore, the expression and the function of seven fucosyltransferases were investigated in normal and cancer pancreatic tissues and in four pancreatic carcinoma cell lines. Transcripts of FUT1, FUT2, FUT3, FUT4, FUT5, and FUT7 were detected by RT-PCR in carcinoma cell lines as well as in normal and tumoral tissues. Interestingly, the FUT6 message was only detected in tumoral tissues. Analysis of the acceptor substrate specificity for fucosyltransferases indicated that alpha(1,2) Fuc-T, alpha(1,3) Fuc-T, and alpha(1,4) Fuc-T were expressed in microsome preparations of all tissues as demonstrated by fucose incorporation into phenyl beta-d-galactoside, 2'-fucosyllactose, N-acetyllactosamine, 3'-sialyl-N-acetyllactosamine, and lacto-N-biose. However, these fucosyltransferase activities varied between tissues. A substantial decrease of alpha(1,2) Fuc-T activity was observed in tumoral tissues and cell lines compared to normal tissues. Conversely, the activity of alpha(1,4) Fuc-T, which generates Lewis a and sialyl-Lewis a structures, and that of alpha(1,3) Fuc-T, able to generate a lactodifucotetraose structure, were very important in SOJ-6 and BxPC-3 cell lines. These increases correlated with an enhanced expression of Lewis a, sialyl-Lewis a, and Lewis y on the cell surface. The activity of alpha(1,3) Fuc-T, which participates in the synthesis of the sialyl-Lewis x structure, was not significantly modified in cell lines compared to normal tissues. However, the sialyl-Lewis x antigen was expressed preferentially on the surface of SOJ-6 and BxPC-3 cell lines but was not detected on Panc-1 and MiaPaca-2 cell lines suggesting that several alpha(1,3) Fuc-T might be involved in sialyl-Lewis x synthesis.


Assuntos
Fucosiltransferases/metabolismo , Isoenzimas/metabolismo , Pâncreas/enzimologia , Idoso , Carboidratos/genética , Carboidratos/imunologia , Epitopos/química , Epitopos/genética , Epitopos/imunologia , Feminino , Fucosiltransferases/genética , Expressão Gênica/genética , Humanos , Isoenzimas/genética , Antígenos CD15/análogos & derivados , Masculino , Pessoa de Meia-Idade , Oligossacarídeos/química , Oligossacarídeos/genética , Oligossacarídeos/imunologia , Pâncreas/química , Neoplasias Pancreáticas/enzimologia , RNA Mensageiro/química , RNA Mensageiro/genética , Antígeno Sialil Lewis X/análogos & derivados , Células Tumorais Cultivadas/química , Células Tumorais Cultivadas/enzimologia
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