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Biomedical subjects

M Soto

Publications and source records attributed to M Soto.

At least 55 records · Page 3Linked to original sources
PubMed1998

[Diagnosis and management of patients with sex differentiation disorders: experience at the Unit of Medical Genetics of the University of Zulia, Maracaibo, Venezuela].

Abnormalities of sexual differentiation (ASD) represent a group of entities, heterogeneous in their etiopathogenesis and clinical manifestations. In order to characterize and analyze the epidemiologic, clinical, endocrine and genetic aspects of patients with ASD consulting UGM-LUZ between 1971-1996, the families that had at least one of its members affected were evaluated. Strict diagnostic criteria to each entity were applied. Cytogenetic, hormonal, radiological, echographic and anatomopathological evaluations were done in each patient. From 391 families, 429 patient consulted with ASD. They represent 5.4% of the patient who consulted to UGM-LUZ in the same period. 214 (50%) patients with definitive diagnosis of ASD were identified to fill the established inclusion criteria. The distribution was the following: 139 with anomalies of the sexual chromosomes; 36 with congenital adrenal hyperplasia; 21 with complete androgen insensitivity syndrome; 14 with mixed gonadal dysgenesis; and 4 with true hermaphroditism. 183 (42.7%) patients with male pseudohermaphroditism and 17 (3.9%) with female pseudohermaphroditism were diagnosed as they did not fulfill the established diagnostic criteria. 15 (3.4%) patients presented ASD associated to a polymalformative syndrome. The ASD are very complex entities, they need the participation of an interdisciplinary team for their diagnosis and management process.

Adolescent↗
PubMed1997

Molecular cloning and analysis of expression of the Leishmania infantum histone H4 genes.

In the present work, we describe the sequence, organization and expression of histone H4 genes in the protozoan parasite Leishmania infantum. The predicted L. infantum histone H4 is a polypeptide of 100 amino acids with a molecular mass of 11.5 kDa. Comparison of the amino acid sequence of Leishmania histone H4 with the rest of histone H4 sequences indicates that this is the most divergent sequence reported to date. The genomic distribution analysis of histone H4 genes indicates that there must be up to seven gene copies. A single size-class histone H4 mRNA of 0.6 kb was detected, whose level dramatically decreases from logarithmic to stationary phase. However, the Leishmania histone H4 mRNAs do not decrease in abundance following treatment with inhibitors of DNA synthesis, suggesting a regulation by a replication-independent mechanism.

Amino Acid Sequence↗
PubMed1997

Analysis of post-transcriptional regulation operating on transcription products of the tandemly linked Leishmania infantum hsp70 genes.

The genomic organization and expression of the hsp70 genes of Leishmania infantum were examined. In the cluster there are at least six copies of the hsp70 genes arranged in a head-to-tail tandem of 3. 8-kilobase repetition units. The hsp70 gene copy (gene 6) located at the 3' end of the tandem has a 3'-untranslated region highly divergent in sequence relative to the 3'-untranslated region of the rest of hsp70 gene copies (genes 1-5). Nuclease S1 protection assays indicated that the steady-state level of the mRNAs derived from gene 6 is about 50-fold more abundant than the transcript level derived from genes 1-5. Nuclear run-on assays showed, however, that all hsp70 genes are transcribed at similar rates. Thus, it is likely that the differences in the steady-state levels of the transcripts from the hsp70 genes should be associated with variations in their processing or maturation rates. While the abundance of the mRNAs derived from hsp70 genes 1-5 is increased by heat shock, the hsp70 gene 6 mRNA level remains unaffected. Our data showed that ongoing protein synthesis is required for the maintenance of the heat inducement, depicting, thus, a post-transcriptional mechanism of positive regulation involving a labile protein factor that would be either induced or activated during heat shock.

Animals↗
PubMed1997

BSD extent, an index for metal pollution screening based on the metal content within digestive cell lysosomes of mussels as determined by autometallography.

The extent of autometallographical black silver deposits (BSD) has been semiquantified at the light microscope in the gills and digestive gland of either control mussels or Zn-polluted mussels after depuration and on exposure to sublethal concentrations of Cu, Zn, and Cd. The BSD extent in the gills and digestive gland of control mussels was much reduced compared to that in other experimental mussels. The extent of BSD in the gills of depurating mussels was reduced at short depuration times due to decreased levels in the abfrontal cells while in the digestive gland it did not change with the depuration period. The extent of BSD in digestive lysosomes of Cu- and Zn-exposed mussels followed a logarithmic pattern in relation to metal concentration increasing with metal concentrations in the digestive gland. However, a reduced extent of BSD was related to the presence of high metal concentrations under Cd-exposure conditions. This is because the great extent of BSD present in the lumen of the digestive tubules was not taken into account to carry out semiquantification, but, however, the chemical analysis measured the Cd content of these BSD. As such, the extent of BSD in digestive lysosomes followed a logarithmic pattern with total metal concentrations in the digestive gland of Cd-exposed mussels. Therefore, the semiquantitative estimation of BSD in the digestive lysosomes could be considered a reliable index to reflect changes in metal bioavailability in sea water.

Analysis of Variance↗
PubMed1997

A region containing repeated elements is associated with transcriptional termination of Leishmania infantum ribosomal RNA genes.

A novel repetitive DNA element has been isolated from the Leishmania infantum genome. The 348 bp long element, designated LiR3, was found to be located downstream from the 3'-end of the ribosomal RNA (rRNA) genes. This LiR3 element has short sequences with potential to form stem-loop structures similar to those of the bacterial rho-independent transcriptional terminators. Given both the structural features and the genomic location of this element we searched for a possible functional implication of these structures in the termination of rRNA transcription. Nuclear run-on assays indicated that indeed there is a transcriptional blockage associated with the LiR3 element. Several chi-like elements, resembling the recombination-promoting sites of Escherichia coli, were identified within the sequences associated with the stem-loop structures. A possible implication of these chi-like elements in rRNA gene conversion events is discussed.

Animals↗
PubMed1997

Complete androgen insensitivity syndrome: clinical and anatomopathological findings in 23 patients.

Complete Androgen Insensitivity Syndrome (CAIS) is a type of Male Pseudohermaphroditism due to a defect in the androgen receptor which is a DNA-binding, transcription-regulating protein whose properties are induced on androgen binding. This defect is caused by mutations of a gene localized in Xq11-12. The objective of this study is the description of the clinical and pathological features of patients with female phenotype, karyotype 46, XY and diagnosis of CAIS. They were referred to Medical Genetic Unit of University of Zulia, Maracaibo, Venezuela, between 1971-1995. The diagnostic criteria and clinical and pathological findings are reviewed. Twenty-three patients fulfilled the diagnostic criteria. Most of patients were 13 years old or older (postpuberal). The main reasons for consultation were: primary amenorrhea, inguinal hernia, familial history of CAIS and fusion of labia minora. Seventeen patients from 5 families were diagnosed with CAIS and only one per family in 6 families. All patients showed a female general phenotype with unambiguously female external genitalia. Gonadal localization (right/left) was as follows: abdominal/abdominal: 65%; abdominal/inguinal: 26%; inguinal/abdominal: 9%. Histologic features of gonads were similar to those in cryptorchid testes of otherwise normal males. There was no evidence of development of gonadal neoplasia. The patients reported here provide a remarkable opportunity to study the molecular genetic characterization that can serve as a primary tool for diagnosis and subsequent therapy.

Adolescent↗
PubMed1997

[Transplacental alloimmunization against specific platelet antigens: prevalence and features in a Chilean population].

BACKGROUND: Neonatal alloimmune thrombocytopenia (NAIT) is a result of fetomaternal incompatibility. Platelet destruction is caused by a maternal antibody directed against a fetal platelet antigen inherited from the father and lacking on the mother's platelets. The incidence and features of transplacental alloimmunization depend on the frequency of expression of platelet specific antigens; which are highly variable among different populations. AIM: To determine the prevalence and characteristics of transplacental alloimmunization in a large group of pregnant women in Chile. MATERIAL AND METHODS: We studied 3,041 samples obtained during the third trimester of gestation. In all samples, anti platelet antibodies were screened by ELISA with platelet membranes fixed to a microtiter plate. Positive samples were further studied for antigenic specificity with the monoclonal antibody specific immobilization of platelet antigens (MAIPA) test. RESULTS: Anti platelet antibodies were found in 261 samples (8.5%). The MAIPA test identified 6 samples with antibodies directed against major platelet membrane glycoproteins, 2 anti GPIb, 2 anti GPIIb/IIIa and 2 anti GPIa/IIa. In four cases, anti HLA antibodies coexisted. Two cases corresponded to well defined platelet antigen systems: one anti HPA-1a and one anti HPA-5b. No clinical evidence of thrombocytopenia of the newborn was detected in all these cases with anti GP antibodies. CONCLUSIONS: A prevalence of platelet specific antibodies of 0.2% with only one anti HPA-1a was detected. These findings are in contrast with those of other populations but in accordance with the low frequency of the HPA-1 b/b phenotype in the Chilean population. The very low incidence of platelet specific antibodies and the lack of association with clinical thrombocytopenia in the newborn, do not support the recommendation of routine antenatal screening to all women in Chile.

Antigens, Human Platelet↗
PubMed1996

Organization, transcription and regulation of the Leishmania infantum histone H3 genes.

The genomic organization and transcription of the genes encoding the histone H3 of the protozoan parasite Leishmania infantum have been studied. It was found that there are multiple copies of the histone H3 genes distributed in chromosomal bands XIX and XIV. The nucleotide sequence of two of the L. infantum H3 genes, each one located in a different chromosome, is reported. Although the nucleotide sequence of the coding region of both genes is identical, the sequence of the 3' untranslated region is highly divergent. It was found also that there exist two different size classes of histone H3 transcripts, each one derived from a different gene, and that they are polyadenylated. The steady-state level of the transcripts dramatically decreases when the parasites enter the stationary phase of growth, suggesting a mode of regulation which is linked to the proliferation status of the cell. Unlike the replication-dependent histones, the L. infantum H3 mRNA levels do not decrease after treatment with DNA synthesis inhibitors. A comparative analysis of the sensitivity of the histone mRNA levels to DNA inhibition in the parasites L. infantum and Trypanosoma cruzi revealed the existence of different control mechanisms in histone expression in these two phylogenetically related protozoan parasites.

Amino Acid Sequence↗
PubMed1996

Tissue-level biomarkers and biological effect of mercury on sentinel slugs, Arion ater.

The present investigation was to determine the extent to which mercury (Hg) provokes measurable effects on the structure of the digestive gland of slugs as well as to relate the extent of these effects to the cell and tissue distribution of Hg. For this purpose, slugs (Arion ater) received various dietary concentrations of Hg (from 0 to 1,000 microg Hg/g food) as chloride for 30 days and were histologically examined every third day. Autometallography was used to demonstrate Hg as black silver deposits (BSD) in paraffin sections. The lysosomes and residual bodies of digestive cells resulted to be the major accumulation sites. In addition, Hg was also evidenced in lipofuscine granules of vacuoles in excretory cells but, however, it was rarely observed within calcium cells. Generally, the extent of BSD increased with dietary Hg concentration and exposure time but, however, it became significant lowered after exposure to 1,000 microg Hg/g food for 30 when the digestive epithelium appeared almost devoid of digestive cells. On the other hand, significant changes were recorded in the quantitative structure of digestive tubules. Mean Epithelial Thickness (MET), Mean Luminal Radius (MLR) and Mean Diverticular Radius (MDR) were recorded as measures of the sublethal biological effect of Hg. MET, MLR/MET and MET/MDR were affected by Hg concentration (C), exposure time (T) and CxT interaction, changes in MET, MLR, MLR/MET and MET/MDR being explained by regression models after logarithmic transformation of the data. In order to explain the nature of the changes in the quantitative structure of the digestive tubules this investigation was complemented with qualitative histological observations. According to them, the excretory activity in digestive cells was initially enhanced. Afterwards, the relative numbers of digestive cells declined until the extreme cases of exposure to 1, 000 microg Hg/g for 27 to 30 days in which the digestive epithelium was mostly comprised of calcium and excretory cells. Concomitantly, some changes took place in blood vessels where Leydig cells became disrupted and the connective tissue layers thickened. Finally, it is suggested to use slugs in soil quality assessment as sentinel organisms ("Slug Watch") in which biomarkers of exposure to metallic pollutants and of biological effect are recorded.

Animals↗
PubMed1996

Autometallographic localization of protein-bound copper and zinc in the common winkle, Littorina littorea: a light microscopical study.

Copper (Cu), zinc (Zn) and calcium (Ca) were demonstrated histochemically by means of conventional stains (rubeanic acid for copper, dithizone for zinc, and cobalt nitrate for calcium) and by autometallography in various tissues of winkles (Littorina littorea) sublethally exposed to either copper or zinc dissolved in sea water. Rubeanic acid and dithizone procedures exhibited poor sensitivity: there was no positive reaction after fixation tissues with Bouin's fixative, and only a weak reaction after ethanol fixation. Autometallography, however, produced a positive reaction with both fixatives in the form of black silver deposits in some key cell types. In winkles not exposed to either copper nor zinc, autometallographically demonstrated metals were found in the connective tissue pore cells, the lysosomes of digestive cells, the basal lamina of the digestive tubule epithelium, and cytoplasmic granules in the epithelial cells of the stomach wall. In addition, in winkles exposed to copper, metal deposits were present in some apical cytoplasmic granules of ciliated cells in the gill epithelium, the mucous secretion of gill mucocytes, and the circulating haemocytes. In winkles exposed to zinc, metal deposits were found in the basal cytoplasmic granules of ciliated cells in the gill epithelium, the mucous secretion of gill mucocytes, the apex and basal lamina of the nephrocytes in the kidney, and the connective tissue layer surrounding the blood vessels. Additionally, calcium was demonstrated histochemically in the cytoplasm of digestive cells, the cytoplasm of the epithelial cells of the stomach wall, the mucocytes of gills, the basal lamina of the kidneys, the haemocytes, the calcium and pore cells of connective tissue, and the oocyte cytoplasm. Metals were not detected by any procedure in sperm cells, in the cytoplasmic granules of oocytes, or in the basophilic cells in the digestive tubules. In conclusion, autometallography is a highly sensitive method and provides an excellent tool to localize protein-bound copper and zinc in molluscan tissues, and its use in combination with conventional histochemical or chemical methods is highly recommended.

Animals↗
PubMed1996

Mapping of the linear antigenic determinants of the Leishmania infantum hsp70 recognized by leishmaniasis sera.

Mapping of antigenic determinants of the Leishmania infantum Hsp70 was studied by analysis of the reactivity of sera from dogs with natural visceral leishmaniasis against a collection of peptides representing overlapping sequences of the Hsp70. Despite the considerable variation in the immune response among individuals three immunodominant regions were revealed encompassing residues 241 to 260 (region I), 435 to 469 (region II) and from amino acid 601 to the carboxyl-terminal end of the protein (region II. Since anti-peptide H17 antibodies purified from a pool of leishmaniasis sera recognized the L. infantum Hsp70 and since they did not react with the homologous Hsp70s from other trypanosomatids, such as Trypanosoma cruzi, Trypanosoma rangeli and Leishmania panamensis, it was concluded that peptide H17 (region II contains an immunodominant B cell species-specific epitope. Our data indicate, however, that the complete recombinant L infantum Hsp70 protein cannot be used as a disease-specific tool for serodiagnosis since it is also recognized by the sera from patients with Chagas' disease.

Amino Acid Sequence↗
PubMed1996

During canine leishmaniasis a protein belonging to the 83-kDa heat-shock protein family elicits a strong humoral response.

By screening of a Leishmania infantum expression library with the serum from a dog affected with visceral leishmaniasis, a cDNA clone with sequence homology to the Hsp83 gene family was isolated. From analysis of the genomic distribution of the cDNA sequence, it was estimated that the L. infantum genome contains 7 Hsp83 genes tandemly organized. The full-length coding region of the Hsp83 gene located at the 5'-end of the cluster was determined. The deduced amino acid sequence of the L. infantum Hsp83 shows a high level of sequence identity with members of the Hsp83's protein family from other eukaryotic organisms. The complete protein (LiHsp83) and 4 subfragments (LiA1, LiB1, LiC1 and LiD1) were expressed in Escherichia coli as recombinant proteins and used as target antigens in FAST-ELISA assays against a collection of sera from dogs with visceral leishmaniasis. Ninety percent of the sera recognized the recombinant LiHsp83, indicating that L. infantum Hsp83 is a potent immunogen during canine leishmaniasis. Serological analysis of the recombinant subfragments identified the LiB1 subfragment, from amino acid 156 to 283, as the immunodominant region of the protein. This region, which is the less evolutionary conserved region of the protein, was recognized by 88% of the visceral leishmaniasis sera. The results suggest that L. infantum Hsp83 and particular protein subfragments may be useful in serodiagnostic assays for canine leishmaniasis.

Animals↗
PubMed1996

Tissue and cell distribution of copper, zinc and cadmium in the mussel, Mytilus galloprovincialis, determined by autometallography.

The localization of metals in selected tissues of metal-exposed mussels was investigated by means of autometallography. Mussels collected from a Zn-polluted site were (a) depurated or, alternatively, (b) exposed to either Cu, Zn or Cd for 41 d. Mussels collected from a clean site were used as experimental reference. Autometallographically demonstrated black silver deposits (BSD), indicating the presence of metals, were observed in gills, (a) in frontal cells and haemocytes of Cu-exposed mussels, (b) in secretory postlateral and abfrontal cells and in endothelial cells of Zn-exposed mussels, and (c) in frontal, postlateral and endothelial cells but mainly in abfrontal cells and haemocytes of Cd-exposed mussels. Autometallography also revealed the presence of BSD in connective tissue brown cells underlying the mantle. Additionally, adipogranular cells of the connective tissue surrounding the gonad follicles were positively stained but no BSD was found in gonad tissue. Scarce BSD were found in the cytoplasmic granules of the stomach wall of control mussels whilst depurating and metal-exposed mussels exhibited BSD lining the apex of the stomach epithelial cells. Contrary to the results previously obtained with marine gastropod molluscs, BSD were not found in the basal lamina of digestive tubules, stomach, and ducts of mussels. Highly conspicuous BSD were observed in digestive cell lysosomes of depurating and metal-exposed mussels. Basophilic cells were always devoid of BSD. Finally, BSD were also found in nephrocyte lysosomes and mineralized concretions.

Animals↗
PubMed1996

During canine viscero-cutaneous leishmaniasis the anti-Hsp70 antibodies are specifically elicited by the parasite protein.

A Leishmania infantum cDNA library was screened with sera from dogs with viscero-cutaneous leishmaniasis. Sequence analysis of a positive clone isolated from the library revealed that it coded for the carboxyl-terminal region of a member of the 70-kDa heat-shock protein family. The full-length sequence of the L. infantum hsp70 gene was determined after isolation of genomic clones. This protein shows a high degree of sequence conservation with the homologous protein from other organisms. To test its antigenicity a recombinant Hsp70 protein fused to the maltose-binding protein was produced in Escherichia coli using the expression vector pMAL-cRI. By FAST-ELISA assays it was observed that while the complete recombinant protein was recognized by 100% of the sera, the 20 carboxyl-terminal amino acids of the protein were only recognized by 30% of those sera. Thus, although a B-cell epitope must be present within the carboxyl terminal end of the protein other antigenic determinant(s) must reside out of this region. The analysis of the cross-reactivity with mouse Hsp70 by Western blotting strongly suggests that the anti-Hsp70 antibodies generated by infection with L. infantum are directed at specific determinants of the L. infantum Hsp70. Thus, our results indicate that anti-Hsp70 autoantibodies are not induced during Leishmania infection.

Amino Acid Sequence↗
PubMed1996

Characterization of the antigenic determinants of the Leishmania infantum histone H3 recognized by antibodies elicited during canine visceral leishmaniasis.

In the present study we show that sera from dogs naturally infected with the protozoan parasite Leishmania infantum contain antibodies that specifically react with the parasite histone H3. Using synthetic peptides covering the complete sequence of the protein we located the linear antigenic determinants within the 40 amino-terminal amino acids of the molecule. In addition to the complete form of the protein (rLiH3), two regions of the Leishmania histone H3 were expressed as recombinant proteins: the rLiH3-Nt fragment containing the 39 amino-terminal amino acids and the rLiH3-Ct fragment containing the 90 carboxyl-terminal residues. Competition experiments using the protein fragment rLiH3-Nt as competitor confirmed that the antigenic determinants of histone H3 are confined to the amino-terminal domain. This domain, which is believed to be exposed on the nucleosome surface, is also the most evolutionarily divergent region of the L. infantum histone H3. Visceral leishmaniasis (VL) sera do not react with mammalian histones, an indication that the anti-histone response elicited during Leishmania infection is triggered by the parasite histone. The results of the prevalence of anti-histone H3 antibodies in canine VL sera together with the sequence-specific characteristics of the amino-terminal region of L. infantum histone H3 indicate that the recombinant protein rLiH3-Nt may be of use for diagnosis of canine VL.

Amino Acid Sequence↗
PubMed1996

Specific serodiagnosis of human leishmaniasis with recombinant Leishmania P2 acidic ribosomal proteins.

The Leishmania P2 proteins have been analyzed as potential tools for the immunodiagnosis of human mucocutaneous and visceral leishmaniasis. Two recombinant Leishmania infantum proteins, rLIP2a and rLip2b, were used. The analysis indicated that the rLiP2a and rLiP2b proteins are recognized by 76% (16 of 21) and 42% (9 of 21), respectively, of sera from patients with mucocutaneous leishmaniasis and by 50% (5 of 10) and 40% (4 of 10), respectively, of sera from patients with visceral leishmaniasis. The Leishmania P2 proteins were engineered to have deletions of particular amino acids from the carboxyl-terminal region in order to avoid cross-reactivity with sera from patients with systemic lupus erythematosus and Chagas' disease, since it is known that this region is the main target of the autoantibodies present in sera from these patients. The results show that while the modified recombinant proteins rLiP2a-Q and rLiP2b-Q, in which the five carboxyl-terminal amino acids had been deleted, maintain the leishmaniasis-specific epitopes, they do not react with sera from patients with autoimmune disease and Chagas' disease. For this reason, and also because the sera from patients with tuberculosis and leprosy, diseases that have to be considered in a differential clinical diagnosis of infectious diseases, do not react with the rLiP2a-Q or rLiP2b-Q protein, we think that the engineered proteins may be considered specific tools for the immunodiagnosis of mucocutaneous and visceral leishmaniasis.

Adolescent↗