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1.
Article in Spanish | MEDLINE | ID: mdl-26273945

ABSTRACT

INTRODUCTION: The doctor-patient relationship is reaching great importance in recent times, is highlighted their importance in areas as varied as satisfaction, compliance, perception of professional competence, the frequency of legal issues relating to malpractice and even the prognosis of the disease or the general health of the patient. OBJECTIVE: To evaluate the doctor-patient relationship from the point of view of residents of emergency unit. METHODS: An observational, descriptive study. The sample consisted of 36 doctors from different areas of the Emergency Rooms of the Hospital de Clínicas- Asunción, Paraguay. The patient-physician relationship was evaluated using an instrument developed by RA Chavarria-Islas et al. with four indicators: Respect, Information, Consent and Dedication. RESULTS: 69.4% of residents have a regular patient-physician relationship; despite the 2.78% has a good relationship, 25% bad relationship and 2.78% a very bad relationship. CONCLUSION: Gaps in doctor-patient relationship were found in this study.. It is interesting to invest greater efforts to enhance the doctor-patient relationship as one of the edges to improve health care, which is vital in emergency care.


Se estudiaron retrospectivamente pacientes con diagnóstico de lupus eritematoso sistémico (LES) de acuerdo a criterios ACR 1982, con nefritis lúpica (NL) durante el período comprendido desde 2005 al 2012 y que fueran sometidos a una biopsia renal repetida. El número total de pacientes con NL atendidos fue de 120, de los cuales 18 (15%) pacientes fueron sometidos a biopsia renal repetida, 18 con 2 biopsias renales y 6 con 3 biopsias. 3 (16,7%) de los pacientes fueron fumadores; 1 (5,6%) poseía antecedentes de DBT previa, 2 (11,1%) poseían antecedentes de HTA; y 3 (16,7%) pacientes tenían obesidad previa. El tiempo de diagnóstico de LES al momento del estudio fue de 96 meses ± 15; el tiempo transcurrido entre la 1° y la 2° biopsia fue de 45 ± 11 meses y el tiempo entre la 2° y 3° biopsia fue de 56 ± 12 meses. Las indicaciones de la biopsia repetida fueron proteinuria en 10 biopsias (41,6%); proteinuria con alteración de la función renal en 2 biopsias (8,3%); proteinuria con sedimento patológico en 8 biopsias (33,3%); y proteinuria con sedimento patológico y alteración de la función renal en 4 biopsias (16,6%). Los cambios histológicos más frecuentes encontrados entre las primeras y las biopsias repetidas fueron: de clase IV a clase III: 2 (8,2%); clase IV a clase IV: 8 (33,3%), clase IV a clase III+V: 2 (8,2%); clase IV a clase IV+V: 3 (12,5%); clase IV a clase V: 2 (8,2%). Los cambios en las biopsias de NL proliferativas con índices de actividad y cronicidad (A/C) fueron: de A a A/C: 7 (29,1%), A/C a A/C: 7 (29,1%). La intensidad de la terapia inmunosupresora aumentó en 79,1%, se mantuvo el tratamiento inmunosupresor en 16.6%. Con respecto al cambio de medicación 7 (20%) pacientes recibieron Ciclofosfamida 1 gr cada 30 días, 9 (26%) Ciclofosfamida 500 mg cada 15 días, 8 (23%) tratamiento de reinducción con Micofenolato mofetil; Rituximab 8 (23%); y 3 (8%) Ciclosporina A. El tratamiento de mantenimiento se realizó con micofenolato mofetil en 23 casos (55%); con azatioprina en 11 (26%) casos; ciclosporina en 3 (7%) oportunidades y rituximab en 5 (12%). En todos los casos se utilizó hidroxicloroquina.


Subject(s)
Attitude of Health Personnel , Clinical Competence/statistics & numerical data , Emergency Service, Hospital/statistics & numerical data , Internship and Residency/statistics & numerical data , Physician-Patient Relations , Clinical Competence/standards , Emergency Service, Hospital/standards , Humans , Internship and Residency/standards , Paraguay
2.
Insect Mol Biol ; 13(1): 73-88, 2004 Feb.
Article in English | MEDLINE | ID: mdl-14728669

ABSTRACT

Anopheles (Nyssorhynchus) darlingi is an important malaria vector in South and Central America; however, little is known about molecular aspects of its biology. Genomic and proteomic analyses were performed on the salivary gland products of Anopheles darlingi. A total of 593 randomly selected, salivary gland-derived cDNAs were sequenced and assembled based on their similarities into 288 clusters. The putative translated proteins were classified into three categories: (S) secretory products, (H) housekeeping products and (U) products with unknown cell location and function. Ninety-three clusters encode putative secreted proteins and several of them, such as an anophelin, a thrombin inhibitor, apyrases and several new members of the D7 protein family, were identified as molecules involved in haematophagy. Sugar-feeding related enzymes (alpha-glucosidases and alpha-amylase) also were found among the secreted salivary products. Ninety-nine clusters encode housekeeping proteins associated with energy metabolism, protein synthesis, signal transduction and other cellular functions. Ninety-seven clusters encode proteins with no similarity with known proteins. Comparison of the sequence divergence of the S and H categories of proteins of An. darlingi and An. gambiae revealed that the salivary proteins are less conserved than the housekeeping proteins, and therefore are changing at a faster evolutionary rate. Tabular and supplementary material containing the cDNA sequences and annotations are available at http://www.ncbi.nlm.nih.gov/projects/Mosquito/A_darlingi_sialome/


Subject(s)
Anopheles/genetics , DNA, Complementary/classification , Gene Library , Saliva/chemistry , Salivary Glands/metabolism , Animals , Brazil , Databases, Genetic , Electrophoresis, Polyacrylamide Gel , Sequence Analysis, DNA
3.
J Insect Sci ; 4: 2, 2004.
Article in English | MEDLINE | ID: mdl-15861218

ABSTRACT

Hexamerins are high molecular-weight proteins found in the hemolymph of insects and have been proposed to function as storage proteins. In previous studies, two Musca domestica hexamerins, designated Hex-L and Hex-F were characterized. Hex-L is synthesized exclusively by the larval fat bodies, is secreted into the hemolymph and likely provides a source of amino acids and energy during metamorphosis. Hex-F synthesis is induced by a proteinaceous meal and occurs only in the adult insect fat bodies. Hex-F also is secreted into the hemolymph and it has been suggested that in females it may be an amino acid reservoir to be used during the final stages of egg formation. Genomic clones containing full-length copies of the genes MdHexL1 and MdHexF1, encoding subunits of the larval and the adult female hexamerin, respectively, were isolated. Complete nucleotide sequences, including the 5'-end untranscribed regions, were determined and analyzed for each of the genes. Comparisons of the conceptual translation products of the cloned genes indicated that MdHexL1 and MdHexF1 are related to the larval serum proteins (LSP) 1 and 2 of Calliphora vicina and Drosophila melanogaster. DNA fragments containing the putative promoters of the two hexamerin genes were compared and cloned into a plasmid vector so as to drive the expression of the GFP reporter gene. The constructs were assayed in vitro in transfected S2 Drosophila melanogaster cells demonstrating that the cloned M. domestica DNA fragments exhibit promoter activity.


Subject(s)
Houseflies/metabolism , Insect Proteins/metabolism , Amino Acid Sequence , Animals , Base Sequence , Cell Line , Gene Expression Regulation , Houseflies/genetics , Insect Proteins/chemistry , Insect Proteins/genetics , Male , Molecular Sequence Data , Oogenesis , Promoter Regions, Genetic
4.
Insect Biochem Mol Biol ; 33(4): 389-95, 2003 Apr.
Article in English | MEDLINE | ID: mdl-12650687

ABSTRACT

The Musca domestica larval hexamerin (MdHex-L) is a hexameric glycoprotein with an apparent native molecular weight of 500 kDa. Seven different cDNAs that encode MdHex-L subunits were cloned and sequenced. Furthermore, amino acid sequences of isolated subunits were determined by the Edman degradation method and compared to the conceptual translation products derived from the cloned cDNAs. The obtained data indicate the existence of multiple forms of MdHex-L subunits and that these multiple forms may be grouped into three categories according to their percentages of nucleotide sequence identity.


Subject(s)
Houseflies/growth & development , Insect Proteins/chemistry , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , Conserved Sequence , DNA Primers , DNA, Complementary/genetics , Hemolymph , Insect Proteins/genetics , Larva , Molecular Sequence Data , Phylogeny , Protein Biosynthesis , Recombinant Proteins/chemistry , Sequence Alignment , Sequence Homology, Amino Acid
5.
Melanoma Res ; 12(6): 529-38, 2002 Dec.
Article in English | MEDLINE | ID: mdl-12459642

ABSTRACT

The presence and characteristics of androgen receptors (ARs) have been described by our group in one human melanoma cell line. We have now investigated their presence in two other human melanoma cell lines, IIB-MEL-LES and IIB-MEL-IAN, as well as in biopsies from human metastatic melanoma. Scatchard analysis revealed a single binding component for both cell lines, the apparent dissociation constant obtained being 15 nM, with a binding capacity of 280 fmol/mg total cell protein, for IIB-MEL-LES cells and 14 nM, with a binding capacity of 206 fmol/mg total cell protein for IIB-MEL-IAN cells. When specificity was assessed, not only androgen and anti-androgen but also non-androgenic compounds were able to compete for [3H]R1881 binding, as seen before. When immunocytochemistry of IIB-MEL-LES and IIB-MEL-IAN cells was performed for ARs, both cell lines were deeply stained in the nucleus, whereas no staining was found for oestrogen or progesterone receptors. Every specimen of melanoma metastases tested for the presence of ARs was deeply stained, and in the majority the intensity of the staining was high. Several hormones and anti-hormones were tested for their ability to affect cell proliferation. In both cell lines, testosterone, dihydrotesterone, oestradiol and progesterone significantly stimulated cell proliferation, and this was reversed by hydroxyflutamide, bicalutamide or tamoxifen.


Subject(s)
Hormones/physiology , Melanoma/metabolism , Melanoma/secondary , Receptors, Androgen/metabolism , Adult , Binding, Competitive , Biopsy , Cell Division/drug effects , Cell Division/physiology , Data Interpretation, Statistical , Dihydrotestosterone/pharmacology , Female , Hormones/pharmacology , Humans , Immunohistochemistry , Male , Melanoma/pathology , Melanoma, Amelanotic/metabolism , Melanoma, Amelanotic/pathology , Melanoma, Amelanotic/secondary , Receptors, Steroid/metabolism , Tumor Cells, Cultured
6.
Rev Med Chil ; 128(7): 721-8, 2000 Jul.
Article in Spanish | MEDLINE | ID: mdl-11050832

ABSTRACT

BACKGROUND: One of the most important factors involved in the pathophysiology of gastroesophageal reflux disease (GERD) is the lower esophageal sphincter rest pressure (LESRP), but these patients can have esophageal motor disorders (EMD). AIM: To assess an association between LESRP and the appearance of EMD in patients with GERD. SUBJECTS AND METHODS: A cross-sectional study was conducted in 229 patients with GERD and 49 healthy controls. Forty five patients with LESRP < 6 mmHg and a mean age of 49 years were assigned to group 1, 128 patients with a LESRP between 6 and 12 mmHg and mean age of 47 years were assigned to group 2, 56 patients with a a LESRP > 12 mmHg and a mean age of 47 years were assigned to group 3 and group 4 was conformed by 49 healthy subjects aged 40 years old. Esophageal manometry was performed using previously published techniques. RESULTS: There was a significant association between LESRP, waves amplitude and the frequency of tertiary waves. CONCLUSIONS: Resting lower esophageal sphincter pressure is inversely proportional to the presence of esophageal motor disorders in patients with gastroesophageal reflux disease.


Subject(s)
Esophageal Motility Disorders/physiopathology , Esophagogastric Junction/physiopathology , Muscle Hypotonia/physiopathology , Adult , Analysis of Variance , Case-Control Studies , Chi-Square Distribution , Cross-Sectional Studies , Esophageal Motility Disorders/complications , Female , Gastroesophageal Reflux/complications , Gastroesophageal Reflux/physiopathology , Humans , Male , Manometry , Middle Aged , Muscle Hypotonia/complications
7.
Insect Mol Biol ; 9(2): 169-77, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10762424

ABSTRACT

Hexamerins are proteins found in high abundance in the haemolymph of larval and adult insects. The expression patterns of the genes encoding the house fly, Musca domestica, hexamerins were determined by Northern analyses using cDNAs as probes. A cDNA, A1, hybridized to a fat body-specific messenger RNA (mRNA) which is detectable in larvae until pupation. Antibodies raised to the larval-specific hexamerin, Hex-L, bind recombinant protein encoded by a 5' rapid amplification of cDNA ends (RACE) product of A1, A2, indicating that the A cDNAs likely represent the genes encoding Hex-L. The F1, F2 and F3 cDNAs, corresponding to genes encoding an adult, female-enriched hexamerin, Hex-F, hybridized with an mRNA isolated from protein-fed females which has a temporal expression profile similar to that observed for the accumulation of Hex-F. Furthermore, expression of the mRNAs hybridizing to the F cDNAs is correlated with the abundance of Hex-F protein during the gonotrophic cycles. The mRNA transcription profiles indicate that the Hex-L and Hex-F genes are regulated in a sex-, tissue- and developmental phase-dependent manner. This stage-specific expression of hexamerins contrasts with the expression patterns of hexamerins seen in other insects. The conceptual translation products of larval hexamerin cDNAs showed identity with larval serum protein 1 (LSP1)-type hexamerins while the deduced products of the female hexamerin cDNAs showed the highest identity with LSP2-type hexamerins. Genomic analyses showed that the larval hexamerin and female hexamerin genes from M. domestica belong to two distinct multigenic families.


Subject(s)
Genes, Insect , Houseflies/growth & development , Houseflies/genetics , Insect Proteins/genetics , Amino Acid Sequence , Animals , Fat Body/chemistry , Female , Gene Library , Hemolymph/chemistry , Larva/genetics , Larva/growth & development , Molecular Sequence Data , Ovary/chemistry , Ovary/growth & development , Pupa/genetics , Pupa/growth & development , RNA, Messenger/isolation & purification , Sequence Homology, Amino Acid , Tissue Distribution
8.
Eur J Clin Nutr ; 54(11): 834-9, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11114677

ABSTRACT

OBJECTIVE: Aging is associated with increased risk of developing anemia and micronutrient deficiencies. Wheat-based staple foods are iron fortified in Chile. We aimed to establish the prevalence and etiology of anemia in apparently healthy free-living elderly subjects. DESIGN AND SETTING: A cross-sectional study was performed in an outpatient clinic of Santiago, Chile. SUBJECTS AND METHODS: A group of 274 subjects (93 men, 181 women)>/=60 y old living at home and apparently healthy was studied. Clinical and anthropometric evaluations and dietary survey were performed. Complete blood count, and status of iron, copper, folate, vitamins B12 and A and C-reactive protein, and erythrocyte sedimentation rate were measured. RESULTS: Prevalence of anemia was 5.4% for men and 4.4% for women. Subjects with inflammatory process had a higher prevalence of anemia (22.2% men, 31.6% women). Abnormal serum retinol (<0.35 micromol/l) was found in 13.7% of men and 15.9% of women. Prevalence of folate deficiency (<7 nmol/l) was 50.5% in men and 33.1% in women. Low serum vitamin B12 (<148 pmol/l) was present in 51.1% of men and 30. 9% of women. Iron and copper deficiencies were infrequent. CONCLUSIONS: Anemia is not prevalent in free-living elderly subjects when iron intake is adequate. Inflammatory process is the main etiology of anemia in this age group. Vitamin A, folate and vitamin B12 deficiencies were found in a significant proportion of the study group. SPONSORSHIP: Sandoz Foundation for Gerontological Research.


Subject(s)
Aging/physiology , Anemia/epidemiology , Inflammation/complications , Iron/blood , Micronutrients/deficiency , Aged , Aged, 80 and over , Anemia/etiology , Anthropometry , Blood Sedimentation , Chile/epidemiology , Cross-Sectional Studies , Diet Surveys , Female , Humans , Inflammation/blood , Male , Micronutrients/physiology , Middle Aged , Prevalence
9.
Cancer Immunol Immunother ; 48(2-3): 100-8, 1999.
Article in English | MEDLINE | ID: mdl-10414463

ABSTRACT

The Lewis(x) (Le(x)) trisaccharide (CD15) linked to proteins and glycolipids is highly expressed on the surface of normal human polymorphonuclear neutrophils (PMN) and several human neoplasias, such as breast and gastrointestinal carcinomas and chronic myeloid leukemias. FC-2.15 is an IgM murine mAb that specifically recognizes Le(x) and has been previously shown to mediate the in vitro lysis of Le(x)(+) cells by human complement. In a phase I clinical trial of FC-2.15, a temporary neutropenia was the main toxicity, and antitumor responses were observed. In order to characterize FC-2.15 further and determine the physiological relevance of Le(x) binding, the reactivity of FC-2.15 on PMN was investigated under several conditions. Flow cytometry revealed a strong reactivity of FC-2.15 with almost 100% of PMN, and Scatchard analysis demonstrated an affinity constant of 5.14 x 10(9) M(-1) and 1.11 x 10(6) antigen sites/cell. In vitro, the binding of Le(x) epitopes by FC-2.15 induced PMN homotypic aggregation, only 28.4 +/- 4.1% remaining as single cells. When PMN and the Le(x)(+) MCF-7 breast cancer cells were co-incubated, FC-2.15 induced heterotypic aggregation. In 51Cr-release assays employing human complement, FC-2.15 lysed 93.4 +/- 7.9% of PMN and 87.8 +/- 10.7% of MCF-7 cells. However, when the effect of FC-2.15 was tested in ex vivo circulating blood, no lytic activity against PMN was detected, whereas MCF-7 cells were still lysed. Blood smears demonstrated that FC-2.15 induced PMN agglutination and heterotypic aggregates when MCF-7 cells were present. A pretreatment of PMN with colchicine impaired PMN agglutination both in vitro (single PMN = 81.15 +/- 4.35%) and in ex vivo circulating blood. In the latter condition, FC-2.15-lytic activity was restored, suggesting that PMN homotypic aggregation by FC-2.15, but not lysis, is dependent on microtubule integrity and that PMN agglutination hinders their lysis. Moreover, when 51Cr-release assays were performed following agglutination, FC-2.15 cytotoxicity was restricted to isolated PMN. It is suggested that crosslinking of Le(x) epitopes by FC-2.15 induces PMN to form homotypic aggregates. It is suggested that the neutropenia observed in FC-2.15-treated patients would be due to PMN agglutination and margination, rather than lysis. In addition, FC-2.15 appears to be able to lyse Le(x)(+) tumor cells in circulation.


Subject(s)
Agglutination , Antibodies, Monoclonal/immunology , Antibody-Dependent Cell Cytotoxicity , Breast Neoplasms/immunology , Lewis X Antigen/immunology , Neutrophils/physiology , Animals , Cell Aggregation , Female , Flow Cytometry , Humans , Mice , Mice, Inbred BALB C , Neutropenia/etiology , Tumor Cells, Cultured
10.
Comp Biochem Physiol B Biochem Mol Biol ; 122(2): 189-98, 1999 Feb.
Article in English | MEDLINE | ID: mdl-10327609

ABSTRACT

The plasma vitellogenin of Bothrops jararaca is composed of two subunits. The larger subunit (160 kDa) is phosphate rich and carbohydrate poor, while the smaller (110 kDa) is highly glycosylated and less phosphorylated. As in other vertebrates, the vitellogenin of B. jararaca is synthesized in the liver under estrogen control. The newly synthesized vitellogenin molecule is a 270 kDa polypeptide. This polypeptide originates the two subunits of the plasma vitellogenin by proteolytic cleavage. In the eggs of B. jararaca six main yolk polypeptides have been detected (113, 107, 104, 72, 27.2 and 20.7 kDa). Using phosphoprotein staining we have shown that the 72 kDa polypeptide is the larger phosvitin so far described in a vertebrate egg yolk. The 107 kDa yolk polypeptide also seems to be phosphorylated, but to a lesser extent than the phosvitin. The 104 kDa vitellin originates from the larger vitellogenin subunit while the 113 kDa vitellin originates from the smaller vitellogenin subunit. Based on these results we propose a general scheme for vitellogenin and vitellin processing in B. jararaca.


Subject(s)
Bothrops/physiology , Egg Proteins/chemistry , Vitellogenins/blood , Vitellogenins/isolation & purification , Animals , Estrogens/pharmacology , Female , Lipoproteins, HDL/analysis , Lipoproteins, HDL/metabolism , Liver/metabolism , Male , Mice , Mice, Inbred BALB C , Protein Biosynthesis , Vitellogenins/metabolism
11.
Cancer Immunol Immunother ; 45(6): 334-9, 1998 Feb.
Article in English | MEDLINE | ID: mdl-9490204

ABSTRACT

FC-2.15 is a murine IgM monoclonal antibody that recognizes breast and colon human carcinomas, chronic myeloid leukemias, Sternberg cells of Hodgkin's lymphoma and some normal cells, such as peripheral polymorphonuclear granulocytes. It has been previously demonstrated that FC-2.15 recognizes the carbohydrate moiety of different glycoproteins. FC-2.15 is able to mediate the in vitro lysis of Ag-2.15+ cells by human complement. In a phase I clinical trial, FC-2.15 induced antitumor responses and reversible neutropenia was its main toxicity. In this work, analysis of epitope specificity has demonstrated that FC-2.15 specifically recognizes terminally exposed Lewis(x) trisaccharide but not sialyl-Lewis(x), Lewis(a), trifucosylated Lewis(y), blood-group antigens A and B, globo H and gangliosides. In polymorphonuclear granulocytes (PMN), myeloid leukemic cells and colon carcinoma T84 cells, Lewis(x) was found to be almost exclusively N-linked to the protein core, whereas in breast carcinoma MCF-7 cells, Lewis(x) appeared to be mostly O-linked. Treatment with neuraminidase increased detection by FC-2.15 in normal PMN, myeloid leukemia cells and T84 cells but not in MCF-7 cells.


Subject(s)
Antibodies, Monoclonal/immunology , Antibodies, Neoplasm/immunology , Breast Neoplasms/immunology , Lewis X Antigen/immunology , Animals , Antibody Specificity , Cell Line , Epitope Mapping , Haptens , Humans , Mice , Mice, Nude , Neoplasm Transplantation , Transplantation, Heterologous , Tumor Cells, Cultured
12.
Endod Dent Traumatol ; 13(3): 146-8, 1997 Jun.
Article in English | MEDLINE | ID: mdl-9550029

ABSTRACT

In this article, we describe the appearance and management of an impacted permanent tooth with severe vestibular root angulation. In addition, the possible origin of this type of root malformation as well as some of their epidemiologic aspects are discussed.


Subject(s)
Incisor/abnormalities , Tooth Root/abnormalities , Tooth, Impacted/etiology , Child , Female , Humans , Maxilla , Tooth Abnormalities/complications , Tooth Abnormalities/surgery , Tooth, Deciduous/abnormalities , Tooth, Impacted/surgery
13.
Insect Mol Biol ; 6(1): 97-104, 1997 Feb.
Article in English | MEDLINE | ID: mdl-9013261

ABSTRACT

During Musca domestica vitellogenesis a protein is preferentially synthesized by the female fat body and accumulates in the haemolymph but not in the ovaries. This protein, designated nonvitellogenic female protein (NVFP), was purified and shown to be a hexamer with an M(r) = 430 kDa, and subunits of M(r) = 70 kDa. The hexamer dissociates into subunits when the pH is elevated from 7.0 to 9.0. Two cDNA clones, F0 and F2, were isolated and analysed. The 2.2 kb F2 clone has an open reading frame that encodes a conceptual translation product that has similarity to the Drosophila melanogaster LSP-2 hexamerin. Recombinant protein from the F2-cDNA is recognized by a specific anti-NVFP serum. The temporal pattern of mRNA expression of the gene represented by the F2 clone follows that determined for the synthesis of NVFP. The data support the conclusion that NVFP is an hexamerin specific to the adult stage of Musca domestica.


Subject(s)
Houseflies/genetics , Insect Proteins/genetics , Amino Acid Sequence , Animals , Antibodies/immunology , Base Sequence , Cloning, Molecular , DNA, Complementary , Escherichia coli , Female , Houseflies/metabolism , Insect Proteins/immunology , Insect Proteins/isolation & purification , Molecular Sequence Data , Rabbits , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/immunology , Recombinant Fusion Proteins/isolation & purification , Sequence Homology, Amino Acid , Vitellogenesis
14.
Photochem Photobiol ; 63(6): 750-4, 1996 Jun.
Article in English | MEDLINE | ID: mdl-8992499

ABSTRACT

Peroxynitrite, the potent oxidant formed by the fast reaction between nitric oxide and superoxide anion, has been suggested to be the reactive intermediate responsible for some of the pathologies associated with an over-production of nitric oxide. In this report, we demonstrate that both nitric oxide and peroxynitrite are formed during infection of the susceptible mouse strain, BALB/c, with Leishmania amazonensis. Nitric oxide was detected as the nitrosyl hemoglobin complex by EPR analysis of blood drawn from mice at 35, 64 and 148 days of infection. The levels of nitrosyl hemoglobin complex increased with disease evolution, which in the murine model used is characterized by skin lesions, ulceration and visceralization of the parasites. Peroxynitrite formation was inferred from immunoreaction of homogenates obtained from footpad lesions in the late stages of the infection with anti-nitrotyrosine antibody; homogenates from parasites drawn from the lesions were also immunoreactive, although to a lesser extent. Analysis of protein homogenates by gel electrophoresis and western blots suggests that peroxynitrite may degrade proteins in vivo, in addition to nitrating them. The results demonstrate that peroxynitrite is formed during murine leishmaniasis and may play a role in the aggravation of the disease.


Subject(s)
Hemoglobins/biosynthesis , Leishmania mexicana , Leishmaniasis, Cutaneous/metabolism , Tyrosine/analogs & derivatives , Animals , Disease Models, Animal , Female , Leishmaniasis, Cutaneous/blood , Mice , Mice, Inbred BALB C , Nitrates/metabolism , Nitric Oxide/biosynthesis , Photobiology , Tyrosine/biosynthesis
15.
Arch Insect Biochem Physiol ; 32(2): 197-207, 1996.
Article in English | MEDLINE | ID: mdl-8785419

ABSTRACT

We describe a method for the purification of ferritin from Musca domestica larval hemolymph. Musca ferritin occurs in hemolymph predominantly as a native protein with molecular weight equal to 550,000 and subunits of 26,000. The average iron content of purified ferritin was determined to be 3,000 +/ 600 iron atoms per molecule. The iron contents of ferritin was heterogeneous; both fully iron loaded molecules and apoferritin are probably present in the Musca hemolymph. The anti-ferritin serum raised in rabbit was able to recognize native ferritin but was not reactive with the protein subunits isolated by SDS-PAGE. The ferritin concentration in hemolymph attains a maximum of 0.28 mg/ml in the wandering stage larvae decreasing to 0.13 mg/ml at the middle of pupal stadium. The ferritin contents of midgut and fat bodies were also determined. Fat body ferritin content is greatly reduced when the feeding larva passes into wandering stage.


Subject(s)
Ferritins/blood , Houseflies/metabolism , Animals , Ferritins/isolation & purification , Hemolymph , Larva , Pupa , Rabbits
16.
Pigment Cell Res ; 8(3): 135-41, 1995 Jun.
Article in English | MEDLINE | ID: mdl-7567789

ABSTRACT

To evaluate the presence of androgen receptors in the human melanoma cell line IIB-MEL-J, a Scatchard plot analysis was performed. Cells in culture revealed a single binding component with an apparent dissociation constant (KD) at 37 degrees C of 11 nM and a binding capacity of 326 fmol/mg protein when measured with [3H]-R1881. Competition analysis revealed an atypical relaxation of specificity, since not only androgen (testosterone, dihydrotestosterone [DHT], R1881) and antiandrogen (hydroxy-flutamide [OH-FLU]) competed for [3H]-R1881 binding, but also estradiol, progesterone, and cortisol at 500-fold excess concentration. Binding of [3H]-estradiol and [3H]-R5020 in the absence of unlabeled DHT were completely suppressed in its presence. Immunohistochemistry of androgen receptor with a monoclonal antibody showed that nuclei were vigorously stained. Different doses of flutamide (FLU) and OH-FLU tested on cultured IIB-MEL-J cells in the presence of serum inhibited significantly cell proliferation in a dose-dependent manner. When cells were incubated with 10 nM DHT and 1% charcoal-adsorbed serum, a significant stimulation of growth that was observed was inhibited by 4 microM OH-FLU. DHT stimulation was completely reversed by the antiestrogen tamoxifen. In addition, male nude mice transplanted with IIB-MEL-J tumor were treated with FLU when tumors were palpable. FLU was effective in diminishing tumor growth and increasing survival rate of the animals. As a conclusion, the presence of functional androgen receptors in these cells has been demonstrated by growth inhibition in vitro and in vivo with antiandrogens, and their atypical nature is suggested by binding cross-reactivity and competition studies.


Subject(s)
Melanoma/metabolism , Receptors, Androgen/metabolism , Androgen Antagonists/pharmacology , Animals , Antibodies, Monoclonal , Antineoplastic Agents, Hormonal/pharmacology , Binding, Competitive , Cell Division/drug effects , Dihydrotestosterone/metabolism , Estradiol/metabolism , Flutamide/analogs & derivatives , Flutamide/metabolism , Flutamide/pharmacology , Humans , Hydrocortisone/metabolism , Immunohistochemistry , Male , Metribolone/metabolism , Mice , Mice, Nude , Progesterone/metabolism , Testosterone/metabolism , Tumor Cells, Cultured
17.
Rev Med Chil ; 122(11): 1289-93, 1994 Nov.
Article in Spanish | MEDLINE | ID: mdl-7659900

ABSTRACT

We studied 36 men and 94 women aged 60 to 85 years, without nutritional deficiencies or illness that could interfere with hematopoiesis, to characterize the normal limits of red and white blood cell counts. Lower limit of hemoglobin normal values were 132 g/l for men and 125 g/l for women. The corresponding figures for MCV were 85 fl and 83 fl for men and women respectively. Lower and upper normal limits for leukocyte count in both sexes were 3.66 and 9.36 x 10(9)/l. Hypersegmented neutrophils existed in 7% of men and 14% of women in spite that folate and vitamin B12 deficiencies were excluded. We concluded that blood cell counts of elderly people have mild deviations of normal values for young adults. Aged humans have a low white cell and bands counts, and elderly women have a higher hemoglobin concentration.


Subject(s)
Blood Cell Count , Age Distribution , Aged , Aged, 80 and over , Female , Hemoglobins/analysis , Humans , Male , Middle Aged , Reference Values , Sex Distribution
18.
Int J Biol Markers ; 9(3): 125-34, 1994.
Article in English | MEDLINE | ID: mdl-7829891

ABSTRACT

FC-2.15 is an IgM monoclonal antibody (MAb) obtained by immunizing Balb/c mice with tumor epithelial cells from a human undifferentiated primary breast carcinoma. FC-2.15 reacts with 93.9% (31/33) of human breast primary tumors, independently of their histology and hormone receptor content. Moreover, FC-2.15 reacts with 79.6 +/- 13.8% (mean +/- SD) of total breast malignant tumor cells and with 88.7 +/- 9.9% of proliferating tumor cells. It recognizes other neoplasia such as colon cancer, squamous carcinoma and melanoma. Among the normal tissues examined, strong cross-reactivity was found with kidney proximal convolute tubules, bone marrow myeloid progeny, peripheral granulocytes and large bowel epithelium. Through Western blots, FC-2.15 recognizes three major bands of Mr 160 kDa, 130 kDa and 115 kDa in membrane extracts of MCF-7 cells grown in nude mice and of human breast carcinoma and three major bands of 250 kDa, 185 kDa and 105 kDa in membrane extracts of peripheral granulocytes. This MAb mediates complement- cytotoxicity against malignant cells, reducing the clonogenic capacity of breast primary tumor cells and MCF-7 cells to 35.6 +/- 41.2% and 11.7 +/- 4.8% of control values respectively, whereas that of normal bone marrow cells is not affected (104.7 +/- 17.4%).


Subject(s)
Antibodies, Monoclonal/immunology , Antibodies, Neoplasm/immunology , Breast Neoplasms/immunology , Neoplasms/immunology , Animals , Antibodies, Monoclonal/isolation & purification , Antibodies, Monoclonal/therapeutic use , Antibodies, Neoplasm/isolation & purification , Antibody Specificity , Antigens, Neoplasm/chemistry , Antigens, Neoplasm/immunology , Biomarkers, Tumor/immunology , Bone Marrow Purging , Breast Neoplasms/therapy , Cytotoxicity, Immunologic , Female , Humans , Immunization , Male , Mice , Mice, Inbred BALB C , Mice, Nude , Molecular Weight , Tumor Cells, Cultured/immunology , Tumor Stem Cell Assay
19.
Comp Biochem Physiol Biochem Mol Biol ; 108(1): 35-9, 1994 May.
Article in English | MEDLINE | ID: mdl-11570397

ABSTRACT

The purified lipophorin of Aedes aegypti (Diptera) is composed of two apolipoproteins: apolipophorin I (M(r)=224,000) and apolipophorin II (M(r)=73,000). The density of lipophorin is constant during the Aedes life-cycle and equal to 1.11 +/- 0.01 g/ml. The amount of lipophorin per animal, during the gonotrophic cycles, increases until 48 hr after blood-feeding and then decreases until there is a new blood intake. The density values and quantification of lipophorin during Aedes aegypti gonotrophic cycle suggest that the adaptation to a higher lipid transport demand during oogenesis in Aedes aegypti is accomplished by increasing the amount of lipophorin in the hemolymph. This response is different from that observed in Musca domestica (Diptera) that does not involve changes in hemolymph lipophorin levels.


Subject(s)
Aedes/metabolism , Carrier Proteins/isolation & purification , Lipoproteins/isolation & purification , Aedes/growth & development , Animals , Carrier Proteins/biosynthesis , Carrier Proteins/chemistry , Eating , Female , Lipoproteins/biosynthesis , Lipoproteins/chemistry , Molecular Weight , Pupa/chemistry
20.
Endod Dent Traumatol ; 9(2): 75-6, 1993 Apr.
Article in English | MEDLINE | ID: mdl-8404700

ABSTRACT

We assessed the bond strength of a glass ionomer cement to dentin that had been in contact with different materials. Flat dentin surfaces in freshly extracted human teeth were covered for 48 h with a 1 mm layer of a variety of materials that are used for temporary filling or root canal sealing. The products were mechanically removed and a glass ionomer cement cylindrical specimen bonded to the dentin surface. After 7-days immersion in 37 degrees C water the tensile bond strength was tested. The results were compared with those on dentin surfaces not in contact with any endodontic material. The statistical analysis showed that none of the materials used interfered with the bonding of the glass ionomer to dentin.


Subject(s)
Dental Bonding , Dentin , Glass Ionomer Cements/chemistry , Root Canal Filling Materials/chemistry , Analysis of Variance , Calcium Hydroxide/chemistry , Calcium Sulfate/chemistry , Camphor/chemistry , Chlorophenols/chemistry , Dental Cements , Drug Combinations , Humans , Hydrocarbons, Iodinated/chemistry , Methylmethacrylates/chemistry , Minerals/chemistry , Polyvinyls/chemistry , Thymol/chemistry , Zinc Oxide/chemistry , Zinc Oxide-Eugenol Cement/chemistry
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