ABSTRACT
INTRODUCTION: Repair of inguinal hernia is one of the most common elective operations performed in general surgery practice. Mesh hernia repair became the gold standard because of its low recurrence rate in comparison with non-tension-free repair. Laparoscopic approach seems to have potential advantages over open techniques, including faster recovery and reduced acute and chronic pain rate. Laparoscopic mesh fixation is usually performed using staples, which is associated with higher cost and risk for chronic pain. Recently, the role of mesh fixation has been questioned by several surgeons. AIM: To evaluate mesh displacement in patients undergoing laparoscopic inguinal hernia repair comparing mesh fixation with no fixation. METHODS: From January 2012 to May 2014, 60 consecutive patients with unilateral inguinal hernia were randomized into two groups: control group--10 patients underwent totally extraperitoneal (TEP) repair with mesh fixation; NO FIX group-50 patients underwent TEP repair with no mesh fixation. Mesh was marked with three 3-mm surgical clips at its medial inferior, medial superior and lateral inferior corners. Mesh displacement was measured by comparing an initial X-ray, performed in the immediate postoperative period, with a second X-ray obtained 30 days later. RESULTS: The mean displacement of all three clips in control group was 0.1-0.35 cm (range 0-1.2 cm), while in NO FIX group was 0.1-0.3 cm (range 0-1.3 cm). The overall displacement of control and NO FIX group did not show any difference (p = 0.50). CONCLUSION: Fixation of the mesh for TEP repair is unnecessary. TEP repair with no mesh fixation is safe and is not associated with increased risk of mesh displacement.
Subject(s)
Hernia, Inguinal/surgery , Herniorrhaphy , Laparoscopy , Postoperative Complications/surgery , Surgical Stapling/adverse effects , Adult , Aged , Elective Surgical Procedures/adverse effects , Female , Herniorrhaphy/methods , Humans , Laparoscopy/methods , Male , Middle Aged , Pain, Postoperative/etiology , Postoperative Complications/prevention & control , Prospective Studies , Recurrence , Surgical Mesh/adverse effects , Surgical Stapling/methods , Treatment OutcomeABSTRACT
Gaucher's disease (GD) is caused by a ß-glucocerebrosidase deficiency, leading to the accumulation of glucocerebroside in the reticuloendothelial system. The prevalence of GD in Tabuleiro do Norte (TN) (1:4000) is the highest in Brazil. The purpose of this study was to present evidence of consanguinity and founder effect for the G377S mutation (c.1246G>A) among GD patients in TN based on enzyme, molecular and genealogical studies. Between March 2009 and December 2010, 131 subjects at risk for GD (GC in dried blood ≤2.19 nmol/h/ml) and 5 confirmed GD patients from the same community were submitted for molecular analysis to characterize the genetic profile of the population. Based on the enzymatic and molecular analysis, the subjects were classified into three categories: affected (n = 5), carrier (n = 20) and non-carrier (n = 111). All carriers were (G377S/wt). Affected subjects were homozygous (G377S/G377S). The identification of a single mutation in carriers and homozygotes from different generations, the history of the community and the genealogy study suggest that the high prevalence of GD in this population may be due to a combination of consanguinity and founder effect for the G377S mutation.
Subject(s)
Gaucher Disease/genetics , Glucosylceramidase/genetics , Adolescent , Adult , Aged , Aged, 80 and over , Amino Acid Substitution , Brazil , Child , Child, Preschool , Consanguinity , Female , Founder Effect , Genetic Association Studies , Glucosylceramidase/deficiency , Humans , Male , Middle Aged , PedigreeABSTRACT
Foram investigadas coinfecções por Leishmania (L.) chagasi, Trypanosoma evansi, Toxoplasma gondii e Neospora caninum em cães. Formas amastigotas de Leishmania spp. foram detectadas pela análise citopatológica de linfonodos em 46.42% (39/84) dos cães. Em um cão macho, adulto, sem raça definida, proveniente de área rural e positivo para Leishmania, foram observadas formas flageladas de T. evansi em esfregaço sanguíneo. Pela imunofluorescência indireta (RIFI), 5.95% (5/84) dos cães foram considerados reagentes para T. gondii, com titulação igual a 64, enquanto que 3.57% (3/84) foram reagentes para N. caninum, com título 50. Entre os animais com leishmaniose visceral, um apresentou resposta sorológica positiva para T. gondii e dois para N. caninum. Todos os cães reagentes para N. caninum eram de área rural e, o predomínio da infecção pelo T. gondii ocorreu em cães da área urbana. Um cão macho, jovem, da zona rural e soropositivo para T. gondii, apresentou mórulas de Ehrlichia spp. na citologia e reação positiva para o vírus da cinomose. Deste modo, mais estudos são necessários para avaliar a epidemiologia dessas infecções na população canina, principalmente com relação aos reservatórios de Trypanosoma spp. nas zonas rurais.(AU)
Co-infections by Leishmania (L.) chagasi, Trypanosoma evansi, Toxoplasma gondii and Neospora caninum in dogs were investigated. Amastigotes forms of Leishmania spp. were detected by cytopathological analysis of lymph nodes in 46,42% (39/84) of dogs. In a male dog, adult, without defined breed, from rural area and positive for Leishmania, were observed flagellated forms of T. evansi in blood smear. By immunofluorescence antibody test, 5,95% (5/84) of dogs were considered reactive to T. gondii, with titer equal to or higher than 1:64, while 3,57% (3/84) were reactive to N. caninum, with titer ≥1:50. Among the animals with visceral leishmaniasis, one showed positive serological response to T. gondii and two for N. caninum. All dogs reactive to N. caninum were from rural area and the predominance of infection by T. gondii was in dogs from urban area. A young male dog from the rural area and seropositive for T. gondii showed Ehrlichia spp. morulae in the cytology and positive reaction for canine distemper virus. Thus, further studies are needed to assess the epidemiology of these infections in canine population, especially with respect to the reservoirs of Trypanosoma spp. in rural areas.(AU)
Subject(s)
Animals , Dogs , Leishmania/growth & development , Toxoplasma/growth & development , Neospora/growth & developmentABSTRACT
Foram investigadas coinfecções por Leishmania (L.) chagasi, Trypanosoma evansi, Toxoplasma gondii e Neospora caninum em cães. Formas amastigotas de Leishmania spp. foram detectadas pela análise citopatológica de linfonodos em 46.42% (39/84) dos cães. Em um cão macho, adulto, sem raça definida, proveniente de área rural e positivo para Leishmania, foram observadas formas flageladas de T. evansi em esfregaço sanguíneo. Pela imunofluorescência indireta (RIFI), 5.95% (5/84) dos cães foram considerados reagentes para T. gondii, com titulação igual a 64, enquanto que 3.57% (3/84) foram reagentes para N. caninum, com título 50. Entre os animais com leishmaniose visceral, um apresentou resposta sorológica positiva para T. gondii e dois para N. caninum. Todos os cães reagentes para N. caninum eram de área rural e, o predomínio da infecção pelo T. gondii ocorreu em cães da área urbana. Um cão macho, jovem, da zona rural e soropositivo para T. gondii, apresentou mórulas de Ehrlichia spp. na citologia e reação positiva para o vírus da cinomose. Deste modo, mais estudos são necessários para avaliar a epidemiologia dessas infecções na população canina, principalmente com relação aos reservatórios de Trypanosoma spp. nas zonas rurais.
Co-infections by Leishmania (L.) chagasi, Trypanosoma evansi, Toxoplasma gondii and Neospora caninum in dogs were investigated. Amastigotes forms of Leishmania spp. were detected by cytopathological analysis of lymph nodes in 46,42% (39/84) of dogs. In a male dog, adult, without defined breed, from rural area and positive for Leishmania, were observed flagellated forms of T. evansi in blood smear. By immunofluorescence antibody test, 5,95% (5/84) of dogs were considered reactive to T. gondii, with titer equal to or higher than 1:64, while 3,57% (3/84) were reactive to N. caninum, with titer ≥1:50. Among the animals with visceral leishmaniasis, one showed positive serological response to T. gondii and two for N. caninum. All dogs reactive to N. caninum were from rural area and the predominance of infection by T. gondii was in dogs from urban area. A young male dog from the rural area and seropositive for T. gondii showed Ehrlichia spp. morulae in the cytology and positive reaction for canine distemper virus. Thus, further studies are needed to assess the epidemiology of these infections in canine population, especially with respect to the reservoirs of Trypanosoma spp. in rural areas.
Subject(s)
Animals , Dogs , Leishmania/growth & development , Neospora/growth & development , Toxoplasma/growth & developmentABSTRACT
Foram investigadas coinfecções por Leishmania (L.) chagasi, Trypanosoma evansi, Toxoplasma gondii e Neospora caninum em cães. Formas amastigotas de Leishmania spp. foram detectadas pela análise citopatológica de linfonodos em 46.42% (39/84) dos cães. Em um cão macho, adulto, sem raça definida, proveniente de área rural e positivo para Leishmania, foram observadas formas flageladas de T. evansi em esfregaço sanguíneo. Pela imunofluorescência indireta (RIFI), 5.95% (5/84) dos cães foram considerados reagentes para T. gondii, com titulação igual a 64, enquanto que 3.57% (3/84) foram reagentes para N. caninum, com título 50. Entre os animais com leishmaniose visceral, um apresentou resposta sorológica positiva para T. gondii e dois para N. caninum. Todos os cães reagentes para N. caninum eram de área rural e, o predomínio da infecção pelo T. gondii ocorreu em cães da área urbana. Um cão macho, jovem, da zona rural e soropositivo para T. gondii, apresentou mórulas de Ehrlichia spp. na citologia e reação positiva para o vírus da cinomose. Deste modo, mais estudos s&at
ABSTRACT
Foram investigadas coinfecções por Leishmania (L.) chagasi, Trypanosoma evansi, Toxoplasma gondii e Neospora caninum em cães. Formas amastigotas de Leishmania spp. foram detectadas pela análise citopatológica de linfonodos em 46.42% (39/84) dos cães. Em um cão macho, adulto, sem raça definida, proveniente de área rural e positivo para Leishmania, foram observadas formas flageladas de T. evansi em esfregaço sanguíneo. Pela imunofluorescência indireta (RIFI), 5.95% (5/84) dos cães foram considerados reagentes para T. gondii, com titulação igual a 64, enquanto que 3.57% (3/84) foram reagentes para N. caninum, com título 50. Entre os animais com leishmaniose visceral, um apresentou resposta sorológica positiva para T. gondii e dois para N. caninum. Todos os cães reagentes para N. caninum eram de área rural e, o predomínio da infecção pelo T. gondii ocorreu em cães da área urbana. Um cão macho, jovem, da zona rural e soropositivo para T. gondii, apresentou mórulas de Ehrlichia spp. na citologia e reação positiva para o vírus da cinomose. Deste modo, mais estudos s&at
ABSTRACT
BACKGROUND: The Epstein-Barr virus (EBV) was used as an agent of B lymphocyte proliferation for subsequent diagnosis of lysosomal storage disease. Due to the constant handling of long-preserved samples in our cell bank, we decided to observe the behavior and then compare cultured and frozen samples for at least one year's cryopreservation. METHODS: Twenty-five samples from healthy individuals were used to assess the possible changes in activity of enzymes ß-galactosidase, ß-glucosidase, α-iduronidase, α-galactosidase, and α-glucosidase. Transmission electron microscopy was used to confirm cell transformation of B lymphocytes into EBV-infected cells, generating lymphoblastoid cell lines. RESULTS: Transmission electron microscopy findings confirmed previous reports in the literature that is, significant and evident morphological changes in the nucleus occur after day 12 and the consequent cell transformation into EBV-infected cells. After thawing and subsequent treatment with the five enzymes utilized, we observed no significant changes in samples cryopreserved for more than one year, as compared to samples cultured for 12 days.
Subject(s)
B-Lymphocytes/enzymology , B-Lymphocytes/virology , Cryopreservation , Hydrolases/metabolism , Lysosomes/enzymology , B-Lymphocytes/metabolism , Cell Line , Cell Transformation, Viral , Herpesvirus 4, Human/metabolism , Humans , Iduronidase/metabolism , Lymphocyte Activation , alpha-Galactosidase/metabolism , alpha-Glucosidases/metabolism , beta-Galactosidase/metabolism , beta-Glucosidase/metabolismABSTRACT
OBJECTIVE: The Epstein-Barr virus (EBV) is utilized as a tool in the study of cellular biology because of its capacity to transform B-lymphocytes. For this reason, EBV is used in conservation of human B-lymphocytes for long periods for subsequent evaluation of lysosomal hydrolase activity. Lymphoblastoid cell lines have several advantages for use over other cell types, such as prompt availability and possibility to develop, characterize and standardize cell banks, to test effects of promising pharmaceutical reagents. The study below presents biochemical data that demonstrate validity of lymphoblastoid cell lines for diagnosis of GM1-gangliosidosis, Gaucher, Fabry and Pompe diseases and mucopolysaccharidosis type I. MATERIALS AND METHODS: Cultures were prepared from peripheral blood, collected from 25 normal subjects and 13 affected individuals. Enzyme activities and immunohistochemistry (IHC) were measured. Activities of enzymes beta-galactosidase, beta-glucosidase, alpha-iduronidase, alpha-galactosidase and alpha-glucosidase were measured before and after cryopreservation for 180 days. Enzymatic activity was measured when transformation was confirmed by IHC. RESULTS: We observed some significant alterations in enzymatic activity of non-cultured cells when compared to others that had been cultured for 12 days and kept frozen for 180 days. CONCLUSIONS: However, these alterations did not invalidate use of the technology of transformation of lymphoblastoid cell lines with EBV, to diagnose the diseases mentioned above, in view of the fact that the cultured cells, before and after freezing, demonstrated similar enzymatic activities.
Subject(s)
B-Lymphocytes , Cryopreservation , Herpesvirus 4, Human , Lysosomal Storage Diseases/diagnosis , B-Lymphocytes/immunology , B-Lymphocytes/metabolism , B-Lymphocytes/virology , Case-Control Studies , Cell Line , Fabry Disease/diagnosis , Fabry Disease/enzymology , Feasibility Studies , Gangliosidosis, GM1/diagnosis , Gangliosidosis, GM1/enzymology , Gaucher Disease/diagnosis , Gaucher Disease/enzymology , Glycogen Storage Disease Type II/diagnosis , Glycogen Storage Disease Type II/enzymology , Herpesvirus 4, Human/immunology , Herpesvirus 4, Human/metabolism , Humans , Iduronidase/immunology , Iduronidase/metabolism , Immunohistochemistry , Lymphocyte Activation/immunology , Lysosomal Storage Diseases/enzymology , Lysosomes/enzymology , Lysosomes/immunology , Lysosomes/virology , Mucopolysaccharidosis I/diagnosis , Mucopolysaccharidosis I/enzymology , alpha-Galactosidase/immunology , alpha-Galactosidase/metabolism , alpha-Glucosidases/immunology , alpha-Glucosidases/metabolism , beta-Galactosidase/immunology , beta-Galactosidase/metabolism , beta-Glucosidase/immunology , beta-Glucosidase/metabolismABSTRACT
This study was designed to evaluate the effect of mycoplasma contamination on acid hydrolase activity and the action of the mycoplasma removal agent (MRA), in cultures of human fibroblasts from individuals with lysosomal diseases. For this purpose, we measured the activity of the b-galactosidase, arylsulphatase B (ASB), hexosaminidase A and a-glucosidase enzymes. The activity of the above mentioned enzymes in fibroblasts contaminated by mycoplasma was measured before and after the addition of the MRA. The results were then compared to the enzymatic activity in contamination-free cultures. Only the ASB enzyme showed significant alteration in activity both in the presence of mycoplasma and MRA. The remaining enzymes did not suffer significant interference by the presence of the two agents. Of the four enzymes tested, three did not suffer significant alterations by the presence of the mycoplasma nor from the MRA. However, the activity measured in the ASB enzyme increased significantly in the presence of mycoplasma and MRA and could lead to a doubtful diagnosis. Therefore, we suggest that contamination should be prevented by using aseptic techniques as well as the MRA in those fibroblast cultures that cannot be discarded.
Subject(s)
Anti-Bacterial Agents/pharmacology , Fibroblasts/microbiology , Hexosaminidase A/analysis , Lysosomal Storage Diseases/enzymology , Mycoplasma/physiology , N-Acetylgalactosamine-4-Sulfatase/analysis , alpha-Glucosidases/analysis , beta-Galactosidase/analysis , Cells, Cultured/drug effects , Cells, Cultured/enzymology , Cells, Cultured/microbiology , Diagnostic Errors/prevention & control , False Negative Reactions , Fibroblasts/drug effects , Fibroblasts/enzymology , Humans , Lysosomal Storage Diseases/diagnosis , Lysosomal Storage Diseases/pathology , Mucopolysaccharidosis VI/diagnosis , Mucopolysaccharidosis VI/enzymology , Mucopolysaccharidosis VI/pathology , Quinolones/pharmacologyABSTRACT
Apolipoprotein CIII (apo-CIII) participates in the regulation of triglyceride-rich lipoprotein metabolism. Several polymorphic sites have been detected within and around the apo-CIII gene. Here, we examined the relationship between apo-CIII SstI polymorphism (CC, CG, GG genotypes) and plasma triglyceride (TG) levels in a group of 159 Japanese individuals living in Southern Brazil. The sample was divided into a group of Japanese descendants (N = 51) with high TG (HTG; >200 mg/dL) and a group of Japanese descendants (N = 108) with normal TG (NTG; <200 mg/dL). TG and total cholesterol levels were analyzed by an enzymatic method using the Labtest-Diagnostic kit and high- and low-density lipoproteins by a direct method using the Labtest-Diagnostic kit and DiaSys Diagnostic System International kit, respectively. A 428-bp sequence of apo-CIII gene was amplified using oligonucleotide primers 5' GGT GAC CGA TGG CTT CAG TTC CCT GA 3' and 5' CAG AAG GTG GAT AGA GCG CTG GCC T 3'. The PCR products were digested with a restriction endonuclease SstI. Rare G allele was highly prevalent in our study population (0.416) compared to Caucasians (0.00-0.11). G allele was almost two times more prevalent in the HTG group compared to the NTG group (P < 0.001). The genotype distribution was consistent with the Hardy-Weinberg equilibrium. There was a significant association between rare G allele and HTG in Japanese individuals living in Southern Brazil as indicated by one-way ANOVA, P < 0.05.
Subject(s)
Apolipoprotein C-III/genetics , Polymorphism, Genetic/genetics , Triglycerides/genetics , Aged , Alleles , Asian People/genetics , Brazil , Coronary Artery Disease/ethnology , Coronary Artery Disease/genetics , Female , Gene Frequency , Genotype , Humans , Japan/ethnology , Male , Middle Aged , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Triglycerides/bloodABSTRACT
Apolipoprotein CIII (apo-CIII) participates in the regulation of triglyceride-rich lipoprotein metabolism. Several polymorphic sites have been detected within and around the apo-CIII gene. Here, we examined the relationship between apo-CIII SstI polymorphism (CC, CG, GG genotypes) and plasma triglyceride (TG) levels in a group of 159 Japanese individuals living in Southern Brazil. The sample was divided into a group of Japanese descendants (N = 51) with high TG (HTG; >200 mg/dL) and a group of Japanese descendants (N = 108) with normal TG (NTG; <200 mg/dL). TG and total cholesterol levels were analyzed by an enzymatic method using the Labtest-Diagnostic kit and high- and low-density lipoproteins by a direct method using the Labtest-Diagnostic kit and DiaSys Diagnostic System International kit, respectively. A 428-bp sequence of apo-CIII gene was amplified using oligonucleotide primers 5' GGT GAC CGA TGG CTT CAG TTC CCT GA 3' and 5' CAG AAG GTG GAT AGA GCG CTG GCC T 3'. The PCR products were digested with a restriction endonuclease SstI. Rare G allele was highly prevalent in our study population (0.416) compared to Caucasians (0.00-0.11). G allele was almost two times more prevalent in the HTG group compared to the NTG group (P < 0.001). The genotype distribution was consistent with the Hardy-Weinberg equilibrium. There was a significant association between rare G allele and HTG in Japanese individuals living in Southern Brazil as indicated by one-way ANOVA, P < 0.05.
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Apolipoprotein C-III/genetics , Polymorphism, Genetic/genetics , Triglycerides/genetics , Alleles , Asian People/genetics , Brazil , Coronary Artery Disease/ethnology , Coronary Artery Disease/genetics , Gene Frequency , Genotype , Japan/ethnology , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Triglycerides/bloodABSTRACT
GM1 gangliosidosis is an autosomal recessive disorder caused by the deficiency of lysosomal acid hydrolase beta-galactosidase (beta-Gal). It is one of the most frequent lysosomal storage disorders in Brazil, with an estimated frequency of 1:17,000. The enzyme is secreted and can be captured by deficient cells and targeted to the lysosomes. There is no effective treatment for GM1 gangliosidosis. To determine the efficiency of an expression vector for correcting the genetic defect of GM1 gangliosidosis, we tested transfer of the beta-Gal gene (Glb1) to fibroblasts in culture using liposomes. Beta-Gal cDNA was cloned into the expression vectors pSCTOP and pREP9. Transfection was performed using 4 microL lipofectamine 2000 and 1.5-2.0 microg DNA. Cells (2 x 10(5)/well) were harvested 24 h, 48 h, and 7 days after transfection. Enzyme specific activity was measured in cell lysate and supernatant by fluorometric assay. Twenty-four hours after transfection, treated cells showed a higher enzyme specific activity (pREP9-beta-Gal: 621.5 +/- 323.0, pSCTOP-beta-Gal: 714.5 +/- 349.5, pREP9-beta-Gal + pSCTOP-beta-Gal: 1859.0 +/- 182.4, and pREP9-ss-Gal + pTRACER: 979.5 +/- 254.9 nmol x h-1 x mg-1 protein) compared to untreated cells (18.0 +/- 3.1 for cell and 32.2 +/- 22.2 nmol x h-1 x mg-1 protein for supernatant). However, cells maintained in culture for 7 days showed values similar to those of untreated patients. In the present study, we were able to transfect primary patients' skin fibroblasts in culture using a non-viral vector which overexpresses the beta-Gal gene for 24 h. This is the first attempt to correct fibroblasts from patients with GM1 gangliosidosis by gene therapy using a non-viral vector.
Subject(s)
Fibroblasts/enzymology , Gangliosidosis, GM1/enzymology , Genetic Vectors , Transfection/methods , beta-Galactosidase/metabolism , DNA, Complementary , Fluorometry , Gangliosidosis, GM1/therapy , Humans , Liposomes , Plasmids/genetics , beta-Galactosidase/geneticsABSTRACT
GM1 gangliosidosis is an autosomal recessive disorder caused by the deficiency of lysosomal acid hydrolase ß-galactosidase (ß-Gal). It is one of the most frequent lysosomal storage disorders in Brazil, with an estimated frequency of 1:17,000. The enzyme is secreted and can be captured by deficient cells and targeted to the lysosomes. There is no effective treatment for GM1 gangliosidosis. To determine the efficiency of an expression vector for correcting the genetic defect of GM1 gangliosidosis, we tested transfer of the ß-Gal gene (Glb1) to fibroblasts in culture using liposomes. ß-Gal cDNA was cloned into the expression vectors pSCTOP and pREP9. Transfection was performed using 4 µL lipofectamine 2000 and 1.5-2.0 µg DNA. Cells (2 x 10(5)/well) were harvested 24 h, 48 h, and 7 days after transfection. Enzyme specific activity was measured in cell lysate and supernatant by fluorometric assay. Twenty-four hours after transfection, treated cells showed a higher enzyme specific activity (pREP9-ß-Gal: 621.5 ± 323.0, pSCTOP-ß-Gal: 714.5 ± 349.5, pREP9-ß-Gal + pSCTOP-ß-Gal: 1859.0 ± 182.4, and pREP9-ß-Gal + pTRACER: 979.5 ± 254.9 nmol·h-1·mg-1 protein) compared to untreated cells (18.0 ± 3.1 for cell and 32.2 ± 22.2 nmol·h-1·mg-1 protein for supernatant). However, cells maintained in culture for 7 days showed values similar to those of untreated patients. In the present study, we were able to transfect primary patients' skin fibroblasts in culture using a non-viral vector which overexpresses the ß-Gal gene for 24 h. This is the first attempt to correct fibroblasts from patients with GM1 gangliosidosis by gene therapy using a non-viral vector.
Subject(s)
Humans , Fibroblasts/enzymology , Genetic Vectors , Gangliosidosis, GM1/enzymology , Transfection/methods , beta-Galactosidase/metabolism , DNA, Complementary , Fluorometry , Gangliosidosis, GM1/therapy , Liposomes , Plasmids/genetics , beta-Galactosidase/geneticsABSTRACT
Epstein-Barr virus (EBV) infection in vitro causes transformation of B cells and generates B lymphoblastoid cell lines (LCLs). These LCLs have been widely used for the diagnostic of several genetic metabolic disorders. However, up to now, efficiency of LCL generation has been based on misleading subjective analysis. In this study, quantitative analyses have been performed to indicate efficiency of B-cell transformation to measuring human lysosomal acid hydrolases associated with: GM1-gangliosidosis type I, Gaucher disease and mucopolysaccharidosis type I. Peripheral blood mononuclear cells were isolated from 13 subjects, and LCLs were produced by culturing them with EBV for 12 days. Activities of the enzymes beta-galactosidase, beta-glucosidase and alpha-iduronidase were measured before and after cryopreservation in liquid nitrogen for 30 days. Efficiency of the B-cell transformation was screened every 4 days by the enumeration of cell proliferation, cell counts and changes in granularity estimated by flow cytometry. We observed the generation of 13 LCLs. Cell transformation was confirmed by the gradual increase of cellular clusters, cell size and granularity. In addition, we determined that the activity of the enzymes mentioned above did not change following cryopreservation. These data suggest that our enumerative approach for screening of EBV-LCLs is efficient for the enzymatic determination of human lysosomal acid hydrolases and may thus replace misleading subjective analyses.
Subject(s)
Cell Transformation, Viral , Cryopreservation , Herpesvirus 4, Human , Iduronidase/metabolism , beta-Galactosidase/metabolism , beta-Glucosidase/metabolism , Adult , B-Lymphocytes/enzymology , B-Lymphocytes/pathology , B-Lymphocytes/virology , Cell Line, Tumor , Cell Proliferation , Gangliosidosis, GM1/diagnosis , Gangliosidosis, GM1/enzymology , Gaucher Disease/diagnosis , Gaucher Disease/enzymology , Humans , Lymphocyte Count , Lysosomes/enzymology , Mucopolysaccharidosis I/diagnosis , Mucopolysaccharidosis I/enzymologyABSTRACT
BACKGROUND: The properties of proton pump inhibitors most investigated are related to peptic diseases and upper gastrointestinal bleeding, but their influence on the healing of sutured gastric incisions has not been assessed. In the present study we evaluated the effect of subcutaneously administered pantoprazole on the healing of sutured gastric incisions in rats. METHODS: Sixty rats were divided into a pantoprazole group and a control group. All rats were submitted to gastric suture in the gastric body and in the gastric fundus and had their gastric pH measured. The pantoprazole group received 20 mg/kg pantoprazole and the control group received 0.9% isotonic NaCl, both subcutaneously t.i.d. Healing analysis was carried out using collagen picrosirius red F3BA staining, and breaking strength was measured on the 4th and 7th postoperative days in all groups. RESULTS: Gastric pH was higher in the pantoprazole group. In the fundus, the pantoprazole group had a higher measurement of breaking strength and a higher proportion of type-I over type-III collagen on the 7th postoperative day. In the body, the pantoprazole group had a higher proportion of type-I over type-III collagen on the 4th and 7th postoperative days. CONCLUSIONS: Pantoprazole given subcutaneously promoted a reduction in gastric acid secretion and was associated with improved healing of the sutured gastric incision in the fundus (squamous epithelium) of rats. These findings suggest that pantoprazole has healing properties in sutured gastric incisions with potential benefits in gastric surgery.
Subject(s)
Anti-Ulcer Agents/pharmacology , Benzimidazoles/pharmacology , Omeprazole/analogs & derivatives , Stomach , Sulfoxides/pharmacology , Wound Healing/drug effects , 2-Pyridinylmethylsulfinylbenzimidazoles , Animals , Collagen Type I/metabolism , Collagen Type III/metabolism , Gastric Acid/chemistry , Hydrogen-Ion Concentration , Male , Omeprazole/pharmacology , Pantoprazole , Rats , Rats, Wistar , Stomach/drug effects , Stomach/pathology , Stomach/surgery , Stress, MechanicalABSTRACT
Recently, an epidemiological association between hepatitis C virus (HCV) infection and type 2 diabetes mellitus (DM) has been reported in several studies, although many of them did not consider known risk factors in the pathogenesis of type 2 DM. The aim of this study was to assess the prevalence of type 2 DM among Brazilian HCV (+) and HCV (-) liver transplant candidates, analyzing known confounding factors for the development of type 2 DM. We conducted a cross-sectional study to evaluate the prevalence of type 2 DM among 106 liver transplant adult candidates, comparing 36 HCV (+) cirrhotic patients with 70 HCV (-) patients who developed cirrhosis from other causes. Type 2 DM was diagnosed after two consecutive fasting glucose values > or =126 mg/dL. The age, sex, and race distribution, severity of liver disease (Child-Pugh score), and family history of DM were similar in both groups, but the mean body mass index (BMI) was higher in the HCV (-) subjects (26.81 +/- 5.29 vs 24.0 +/- 4.71, P < .01) Most of the patients were Caucasians (70.75%). Type 2 DM was detected in 36.11% of HCV (+) group and in 25.71% of the HCV (-) (P = .27). A multivariate analysis revealed that family history of DM was the only significant independent predictor for DM (odds ratio = 2.55, 95% CI = 1.03 to 6.31, P = .04). In conclusion, our study did not show an association between HCV infection and Type 2 DM in Brazilian liver transplant candidates. It confirmed that the family history of DM was a determinant factor for the development of type 2 DM.
Subject(s)
Diabetes Mellitus, Type 2/epidemiology , Liver Failure/epidemiology , Liver Transplantation/statistics & numerical data , Body Mass Index , Brazil/epidemiology , Cross-Sectional Studies , Diabetes Mellitus, Type 2/etiology , Female , Humans , Liver Failure/complications , Liver Failure/etiology , Male , Middle Aged , Prevalence , Waiting ListsABSTRACT
New-onset diabetes melittus (NODM) is a serious complication following transplantation. Recent studies suggest an association between hepatitis C virus (HCV) infection and DM both in nontransplant settings as well as after liver transplantation (LT). The aim of this study was to assess the prevalence of NODM among Brazilian LT recipients, analyzing possible risk factors including HCV infection. We conducted a cross-sectional study to evaluate the prevalence of NODM in 82 LT recipients with a posttransplant follow-up > or =1 year including 29 HCV-positive patients and 53 with other causes for liver disease. Patients were considered to meet the criteria for DM if they had two consecutive fasting glucose values > or =126 mg/dL or if they were taking insulin or oral hypoglycemic agents at the time of the study. The overall prevalence of NODM was 18.29% with a median interval of 20 months between LT and diagnosis of DM. The age, sex, and race distribution, immunosuppressive regimen, number of rejection episodes treated with pulse therapy, and family history of DM were similar in both groups. However, the frequency of BMI > or = 30 in the pre- and posttransplant periods was higher among patients who developed NODM (P = .02). Upon multivariate analysis of the entire cohort, HCV infection was the only significant predictor of NODM (OR = 4.31, CI = 1.17 to 15.84, P = .02). In conclusion, our study confirmed an association between HCV infection and NODM among Brazilian liver transplant recipients, suggesting that HCV infection may have a potential role in the pathogenesis of posttransplantation DM.
Subject(s)
Diabetes Mellitus/epidemiology , Hepatitis C/epidemiology , Liver Transplantation/statistics & numerical data , Brazil/epidemiology , Cross-Sectional Studies , Female , Follow-Up Studies , Hepatitis C/complications , Hepatitis C/surgery , Humans , Male , Prevalence , Risk Factors , Time FactorsABSTRACT
Living donor liver transplantation (LDLT) for children and adults has gained widespread acceptance due to the severe organ shortage. LDLT provides potential recipients with timely transplantation, but this procedure engenders a potentially significant risk to the donor. This study analyzed medical, functional, and psychological donor outcomes after LDLT. Nineteen donors (mean age 33.9 +/- 12 years), who underwent hepatectomy for LDLT (13 right lobectomy for adult LDLT) from March 1998 to November 2002, were interviewed at a median of 13 months after donation (range, 2 to 58 months). According to the Clavien System classification, major complications occurred in three donors (16%), and minor in four (21%). The mean length of hospital stay was 5.7 +/- 1.6 days. Five patients (27%) needed rehospitalization. Complete recovery was achieved at a mean time of 8.5 +/- 3.5 weeks. All 19 donors were able to return to predonation activities. The donor's relationship to the recipient and to their families was improved after donation in all cases; 12 (63%) cited a positive psychological impact on their lives. About 90% would donate again and 84% would recommend donation to someone contemplating it. In conclusion, all donors are alive and well after donation and were able to return to their predonation occupation. Most of them felt that this experience changed their lives for the better and would donate again. Donor safety and quality of life should remain the priority in all donation processes.
Subject(s)
Liver , Living Donors/psychology , Quality of Life , Adolescent , Adult , Female , Hepatectomy/methods , Humans , Interviews as Topic , Liver Transplantation , Male , Middle Aged , Time Factors , Tissue and Organ Harvesting/methodsABSTRACT
OBJECTIVE: To determine the prevalence of malnutrition among liver transplant (LT) candidates. MATERIALS AND METHODS: A prospective study evaluated 219 adult LT candidates including 141 men and 78 women. Cholestatic disease was present in 21 (Child: A = 1, B = 11, and C = 9) and noncholestatic disease in 198 (Child: A = 12, B = 93, and C = 93. The mean age was respectively 45.6 and 46.5 years. Anthropometric and biochemical assessments were performed for statistical analysis using Student t test (P <.05). RESULTS: In the noncholestatic group, 41.5% were obese according to keep a body mass index (BMI); 61.6% were depleted according to adequacy of tricipital skin fold (%TSF); and 71.1% were above normal levels for generalized adipose reserve (%F). In terms of adequacy of mid-upper arm muscle circumference (%MMC), 58% were depleted and 50.5% were depleted for the current body weight/usual body weight (%CBW/UBW). Otherwise 52.2% of current body weight/ideal body weight (%CBW/IBW) values were above normal. Serum albumin was below normal in 64.9% of cases. In the cholestatic group 62% were normal for BMI; 66.7% were depleted for %TSF; 77.8% were above normal for %F. As to %MMC, 47.6% were depleted and 47.6% were depleted for %CBW/UBW. Otherwise 47.6% were above normal weight for %CBW/IBW. Serum albumin was below normal in 53.9% and %MMC values showed statistically significant differences (P =.02) when compared with Child B and C in the noncholestatic group, as well as %F (P =.01) and serum albumin (P =.0002) in the cholestatic and noncholestatic groups. Serum albumin values also showed statistically significant differences (P =.0004) when noncholestatic Child B and C patients were compared. CONCLUSION: Patients with cholestatic disease were more affected by calorie depletion compared to noncholestatic patients who were more affected by protein depletion.
Subject(s)
Liver Transplantation , Malnutrition/epidemiology , Waiting Lists , Adult , Body Weight , Cholestasis/epidemiology , Cholestasis/surgery , Female , Humans , Liver Diseases/epidemiology , Liver Diseases/surgery , Male , Malnutrition/physiopathology , Nutritional Status , Prevalence , Reference Values , Retrospective Studies , Serum Albumin/analysisABSTRACT
Abnormalities in the reproductive function and sexuality, which are common among women with advanced liver disease, may reverse after successful liver transplantation (LT). To analyze reproductive function and sexuality in women who underwent successful LT, we interviewed 28 recipients (mean age 44.17 +/- 13.6 years old) at a median posttransplant survival of 36.5 months (range, 6 to 110 months), with good graft function and obeying regular follow-up at our institution. In addition to medical records, all subjects answered a questionnaire on their menstrual pattern, sexual activity, contraceptive practice, pregnancy, and sexuality domain. Nineteen of 22 patients in the child bearing age (86.4%) recovered menstrual function at a median of 1 month after LT (range, 1 to 7 months). Twenty of 28 recipients (71.4%) were sexually active. The most frequent contraceptive practices were barrier methods and tubal ligation. There were four successful pregnancies (one twin) in three patients; five healthy babies were delivered. Overall, 70% of sexually active patients indicated satisfaction with their relationship, 75% had weekly intercourse, and 70% experienced orgasm with intercourse. Eighty percent expressed a desire to receive information concerning sexuality. In conclusion, LT has a positive impact on sexuality and reproductive function in female recipients. It would desirable that LT programs included information regarding these issues for this population.