Mucopolysaccharidosis type VII (beta-glucuronidase deficiency): a chronic variant with an oligosymptomatic severe skeletal dysplasia.

We report on a 20-year-old male with a beta-glucuronidase (GUSB) deficiency mucopolysaccharidosis. He had pectus carinatum, gross thoracic kyphoscoliosis, and hip dysplasia, a picture which became conspicuous after age 4 years. Hepatosplenomegaly, herniae, corneal clouding, and neurological abnormalities were absent. Although he had Alder-type granulations in his polymorphonuclear leukocytes, the urine did not contain a significant excess of mucopolysaccharides. Electron microscopic examination of skin and gingival biopsies, leukocytes, and cultured skin fibroblasts showed numerous single membrane-limited vacuoles either empty or filled with fibrillogranular material; this last tissue did not contain metachromatic granules. Radiographs demonstrated a distinct spondyloepiphyseal dysplasia in which the most striking changes were confined to the thoracic spine (flattening and collapse in T7, T8 and T10 vertebral bodies) and to the femoral capital epiphyses (irregularities and fragmentation). The activity of GUSB in the patient's serum, leukocytes, and fibroblasts was severely decreased; the GUSB activity in the serum and leukocytes from the parents and 2 asymptomatic sibs was subnormal. Immunoblot analysis showed very low levels of cross-reactive material towards anti-GUSB antiserum in the patient's leukocyte and fibroblast extracts. This patient was more severely affected in his skeleton than other described patients with an oligosymptomatic chronic form. This case broadens the clinical and biochemical picture associated with GUSB deficiency and may represent a new variant of the disease.

Barth's syndrome-like disorder: a new phenotype with a maternally inherited A3243G substitution of mitochondrial DNA (MELAS mutation).

An Argentine male child died at 4.5 years of age of a lethal mitochondrial disease associated with a MELAS mutation and a Barth syndrome-like presentation. The child had severe failure to thrive from the early months and for approximately two years thereafter. In addition, the patient had severely delayed gross motor milestones, marked muscle weakness, and dilated cardiomyopathy that progressed to congestive heart failure. He also had persistently elevated urinary levels of 3-methylglutaconic and 2-ethylhydracrylic acids and low blood levels of cholesterol. Detailed histopathologic evaluation of the skeletal muscle biopsy showed high activity of succinate dehydrogenase, a generalized decrease of COX activity, and abundant ragged-red fibers. Electron microscopic studies revealed multiple mitochondrial abnormalities in lymphocytes and monocytes, in the striated muscle, and in the postmortem samples (muscle, heart, liver, and brain). Biochemical analysis showed a pronounced and constant lactic acidosis, and abnormal urinary organic acid excretion (unchanged in the fasting and postprandial states). In addition, in CSF there was a marked increase of lactate and beta-hydroxybutyrate (beta-HOB) and also a high systemic ratio beta-HOB/acetoacetate. Enzymatic assay of the respiratory chain in biopsied muscle showed 10% of complex I activity and 24% of complex IV activity compared with controls. Molecular studies of the mitochondrial genome revealed an A to G mutation at nucleotide pair 3243 in mitochondrial DNA, a well-known pathogenetic mutation (MELAS mutation) in all the patient's tissues and also in the blood specimens of the probands mother and sibs (4 of 5). The diagnosis of MELAS mutation was reinforced by the absence of an identifiable mutation in the X-linked G4.5 gene of the propositus. The present observation gives additional evidence of the variable clinical expression of mtDNA mutations in humans and demonstrates that all clinical variants deserve adequate investigation to establish a primary defect. It also suggests adding Barth-like syndrome to the list of phenotypes with the MELAS mutation.

Electrolyte distribution in toad sciatic nerve and spinal cord.

The distribution of Na+, K+ and water was studied in spinal cord, sciatic nerve and sartorius muscle of the toad, Bufo arenarum (Hensel). Electrolyte assays and sodium washout curves were made. In sartorius muscle, extracellular water was estimated to account for 18.9% of total tissue weight and the intracellular concentrations of Na+ and K+ were 18.4 and 147.9 mEq/kg of intracellular water respectively. In spinal cord extracellular water was 17.2%, intracellular Na+ was 40.6 mEq/kg of water, and intracellular K+ was at a concentration of 134.5 mEq/kg. Interpretation of washout curves in sciatic nerve could not be as simple as in other tissues due to the connective sheath, and therefore electron micrograph measurements had to be made in order to estimate how much of the total tissue weight could be attributed to the sheath. Assuming that it had a water and electrolyte composition similar to that of plasma, the sheath corresponded to 22.3% of the nerves and contained a proportional fraction of Na+ and K+. These calculations left an 'excess' of Na+ and water to be distributed among the remaining components of sciatic nerve. Application of the pyroantimoniate histochemical technique for Na+ determination disclosed a large amount of precipitate among myelin lamellae both in sciatic nerve and in spinal cord. This might explain at least in part the distribution of the 'excess' Na+ in both nervous tissues.

[Phenotypic expression variation of isovaleric acidemia in Argentinian patients. A long term follow-up]. / Variación en la expresión fenotípica de la acidemia isovalérica en pacientes argentinos. Observaciones de un prolongado seguimiento.

In this paper we discuss the first five Argentinean patients presenting isovaleric acidemia (IVA), an alteration of leucine catabolism due to a genetic defect of isovaleryl-CoA dehydrogenase. Belonging to unrelated families, one from native (H. Fam.) and the other from Italian ancestry (M. Fam.); the patients presented the clinical pattern highly suggestive of the disease: they were siblings, had disease-free intervals, vomiting, ketoacidosis crises, "sweaty feet" odor and progression of the neurologic involvement from somnolence and stupor to profound coma. In the four children of H. Fam. the disease had a late but severe beginning; one of the girls died (N.H.). The boy from M. Fam. presented a neonatal form of clearly benign course. The disease was confirmed by gas-chromatography (GC) of volatile acids in serum and also by the typical urinary acid GC-profiles (Fig. 1, A and B); the isovalerylglycine quantitative evaluation in urinary samples collected during crises is shown in Table 1. The morphological findings in liver and brain of N.H. showed at the ultrastructural study, an extensive fatty degeneration and greatly marked mitochondrial alterations in the liver and edema, neuronal karyorrhexis and karyolysis in the brain (Fig. 2). The therapeutic protocol based on a low leucine or low protein diet and use of glycine is described. The evolutionary follow up, more than 10 years for the first case, showed a normal mental development in three of them and retardation in the first child of H. Fam., who had a late diagnosis. IVA is still valuable as a paradigm in the acquisition of a highly clinical suspicion and for its introduction in the study of genetic organic acidemias.

[Hepatic glycogen synthetase deficiency or glycogen storage disease-zero. Mild phenotype with partial enzymatic defect]. / Deficiencia en glucógeno sintetasa hepática o glucogenosis tipo cero. Fenotipo menos severo con defecto enzimático parcial.

Since the original description 26 years ago, of the hepatic glycogen synthetase deficiency, only one more case was reported in 1977. We present the studies carried out on an Argentine boy of Italian ancestry who at age 21 months, showed signs of hepatic deficiency with mild clinical symptoms which contrasted with a remarkable fatty liver degeneration. A totally atypic reaction to fructose overload (Table 1, Fig. 1) was the first key to the diagnosis. Glucose levels were not significantly modified by glucagon after 12-hours fasting, but it did increase the glycemia, with decrease of lactate and alanine 3 hours after-meal (Fig. 2a, b). The 24-hours metabolic profile showed fasting hypoglycemia, hyperketonemia, low alanine concentrations and mild lactatemia and hyperglycemia and a net post-prandial increase of lactate (Fig. 3). This profile when reduced to 14 hours, 12-fasting hours and 2-postprandial hours (Fig. 4), revealed similar alterations in an asymptomatic younger brother. The development of the investigation led to a second hepatic biopsy which confirmed hepatic steatosis and to an ultrastructural study, which showed subcellular alterations in the liver and also in muscle (Fig. 5). Moreover low content of hepatic glycogen was observed along with glycogen synthetase activity between 20-25% that of controls, being normal the enzyme activity in muscle and fibroblasts cultured from a skin biopsy (Table 2). The clinical pattern mainly without hypoglycemia, convulsions and/or mental retardation and a normal height and body mass development, allowed us to postulate that this Argentine report would be a mild variant of the disease formerly described and would be correlated with a partial deficiency of the hepatic glycogen synthetase.

[Salla disease (sialuria, Finnish type). New clinical presentation in the 1st Argentine report]. / Enfermedad de Salla (sialuria, tipo Finlandés). Nueva variante clínica en un primer reconocimiento Argentino.

Studies in three sibs from an Argentine family, aged 29, 18 and 9 years, suffering from a severe neurological disease, revealed in the two older brothers (the third died), ultrastructural changes in cellular vacuolization in diverse peripheral tissues (conjunctival, gum and skin biopsies) and in blood lymphocytes. These data were suggestive of mucopolysaccharidosis, mucolipidosis or glycoproteinosis. However, the activity of lysosomal enzymes, the excretion of mucopolysaccharides and oligosaccharides reactive to orcinol, as well as the search for aspartylglucosaminuria gave normal values. The main biochemical finding was the detection of a substantial urinary increase of a unique resorcinol-positive compound, which by thin-layer chromatography was identified as N-acetylneuraminic acid (NANA-Free) and when quantified by the thiobarbituric acid method previously passed through a gel filtration column (Sephadex G-15) or through ion exchange resins, showed a NANA-Free concentration about 15 times higher than in controls of similar age (Table 2). The ultrastructural findings (Figs. 3-5), the hypersialuria and the present clinical state of these patients (Table 1, Figs. 1, 2) were compatible with Salla disease, a rare lysosomal storage disease originally observed in Finland. The precocity and severity of the neurological damage in our patients were evident since birth and without maturing accomplishments in their first years, contrary to the progressive neurological regression described for the classical syndrome. Based on these facts we suggest that the Argentine patients would constitute a new clinical form of Salla disease.

[Differential expression of mucin carbohydrates in human endometria]. / Expresión diferencial de los carbohidratos de mucina en endometrios humanos.

The use of Lectins to identify oligosaccharides in mucin substances has been increased by the role played by cell surface carbohydrates in invasion and metastasis processes. We studied in this work normal endometrial tissue, with benign and malignant entities in search for the presence of the Galactose beta 1-3 N Acetylgalactosamine(Gal beta 1-3 GalNAC alpha and Galactose beta 1-3 N Acetylgalactosamine (Gal beta 1-3 alpha and beta) using the Lectins: Agaricus bisporus (ABL) and Arachis hipogea (PNA) respectively. The specific control were baths with galactose for PNA and with porcine stomach mucin for ABL. The use of these two Lectins allowed to differentiate substances bonded or non bonded to Sialic Acid, since PNA fails to label when the oligosaccharide is bonded to this acid Sialic. Significant differences were noticed on the bonding patterns of both Lectins on tissues with benign, malignant and normal entities. In this latter case the labelling was always continuous in both Lectins whereas it was irregular in the carcinoma.

[Granulosa cell tumors: structural and ultrastructural analysis]. / Tumores de células de la granulosa: análisis estructural y ultraestructural.

UNLABELLED: These ovarian neoplasm derive from the ovarian stromal component constituting around the 5 to 12% of all ovarian tumors. OBJECTIVE: To examine the histopathological and ultrastructural morphologic characteristic of the neoplastic cells and the patognomonic element of these tumors: Call Exner's Bodies MATERIALS AND METHODS: The materials corresponded to 2 women of 52 and 55 years. The syntomatology was abdominal tumor that went in increase. The materials were fractioned for the histopathological conventional study and for ultrastructural analysis. For this last one, they were fixed in Karnovsky, refix in osmio and included in Araldita. RESULTS: By means of the different observations it was determined in both cases the nuclear atipia, indentations nuclei and prominent nucleoli. In one of the cases the presence of Bodies of Call-Exner was detected, and ultrastructurally was compound by whirled fibrils and amorphous material, with dense structures electron to its around. It was interest the infiltrated of plasmatic cells around the tumors cells. These neoplasms are of interest due to their impredictible behavior and to the hormonal production that can originate alterations in other organs of the genital apparatus.

Creatine kinase and lactate dehydrogenase levels as potential indicators of Trypanosoma cruzi infectivity and histotropism in experimental Chagas' disease.

The results of the present study reveal an early increase in activity levels of creatine kinase and lactate dehydrogenase in the plasma of mice infected with Trypanosoma cruzi strains K-1, X-1, and Tulahuen as compared with uninfected control mice. An increase in creatine kinase activity was detected earlier in K-1- and X-1-infected mice than in Tulahuen-infected mice. Moreover, an increase in lactate dehydrogenase activity occurred at 1.5 days after infection with the X-1 and Tulahuen strains and at 3.5 days after infection with the K-1 strain. Generally, the highest activity levels were found in the plasma of mice infected with the most virulent and lethal Tulahuen strain as compared with the less virulent and nonlethal K-1 and X-1 strains. A significant decrease in creatine kinase levels occurred later in the tissues than in the plasma of K-1- and X-1-infected mice but did not vary significantly in any of the tissues from Tulahuen-infected mice. Similarly, the specific activity of lactate dehydrogenase in tissues from K-1- and X-1-infected mice dropped at a later stage than did the activity in plasma, but infection with the Tulahuen strain caused an earlier reduction in the activity of lactate dehydrogenase in the heart and skeletal muscle. The activity levels of both enzymes in plasma and tissues showed a linearly negative and statistically significant correlation. The present study reveals that levels of creatine kinase and lactate dehydrogenase activity in plasma could be early indicators of and suitable tools for monitoring of the infectivity of these strains of T. cruzi and might reflect their inherent histotropism during experimentally acute Chagas' disease.

Effect of estrogen and antiestrogen therapy in rats bearing mesenchymal tumors.

We applied both hormonal and antiestrogen treatment in female Wistar rats to analyze the estrogen dependence of the growth of sarcomas induced with 9,10-dimethyl-1,2-benzanthracene. Animals bearing tumors of 10 mm in diameter were divided at random into five groups and submitted to different treatments during 24 weeks. The treatment with ovariectomy and tamoxifen in tumor-bearing animals resulted in tumor growth suppression and prolonged survival by a protection against the lethal tumor. On the other hand, the estrogen treatment exerted an adverse effect showing a faster growth of the tumors and a great decrease in survival. In summary, the antiestrogen treatment can have an antitumor effect in mesenchymal tumors, possibly by modifying the immunological status of the host.

Synoviorthesis with 32P-colloidal chromic phosphate in rheumatoid arthritis--clinical, histopathologic and arthrographic changes.

Synoviorthesis was performed in 217 joints from 111 patients suffering from different stages of rheumatoid arthritis (RA). 32P-colloidal chromic phosphate was employed, with an average dose from 6 mCi for large joints (knees) to 0.3 mCi for small peripheral joints such as average dose from 6 mCi for large joints (knees) to 0.3 mCi for small peripheral joints such as the MCP or PIP joints. Satisfactory clinical results were observed in 84% of the cases and no significant side effects resulted after a follow-up period from 1 to 10 years. Striking effects after treatment were observed through histopathological studies (light and electron microscopy) and the use of contrast arthrography. We concluded that radioactive synovectomy with 32P-chromate is a very useful method for the local treatment of RA.

Relationship between immune state and tumor growth rate in rats bearing progressive and non-progressive mammary tumors.

Impaired immune responses occur frequently in cancer patients or in tumor-bearing animals, but the mechanisms of the tumor-induced immune defects remain poorly understood. The aim of the present study was to determine the relevance of the immune system in the control of tumor growth. We have developed a model of progressive and non-progressive mammary tumor, chemically induced in female Wistar rats. In this model we evaluated the development of an immune response after immunization of rats bearing progressive and non-progressive tumors with a non-related antigen, such as sheep red blood cells. We also studied the activation state of peritoneal macrophages from animals bearing tumors by evaluating the production of free radicals. Our findings indicated that the cell-mediated immunity in rats bearing progressive tumors fails to respond to heterologous antigen in vivo, as demonstrated by a negative delayed-type hypersensitivity reaction, and is accompanied by minor nitric oxide production by peritoneal exudate cells as well as a lower capacity for macrophage activation. The study of non-progressive tumor-bearing rats indicated that the cell-mediated immune response was intact and an activated state of macrophages was found in vivo. The results described in this paper should be taken into account when therapies based on cancer vaccines are chosen for the treatment of cancer.

Induction of organ-specific antigens of the rabbit male accessory glands by injection of testosterone or gonadotrophin.

Highly specific antiserums for two antigens of rabbit male accessory glands, called FII and FV, were used to study the effect of hormonal treatment on their appearance during postnatal development. Single or double serial injection of testosterone, or human chorionic gonadotrophin, produces an increase in accessory gland and testis weight, and at the same time induces the synthesis of a detectable amount of specific antigens. This was proved by using groups of animals 4 and 6 1/2 weeks old. The antigens were consistently absent in accessory glands from normal and control animals of the same age. The histologic characteristics of maturation were also observed. Ultrastructural studies revealed an increase in number of microvilli supranuclear vaculoes with secretory content and well developed rough endoplasmic reticulum. These changes were more easily detected in young animals treated with chorionic gonadotrophin. The significance of these findings is discussed in relation to the presence of specific carriers (cytosols).

Vascular endothelium express CS-1 fibronectin in allergic contact dermatitis.

BACKGROUND: Allergic contact dermatitis (ACD) is a common human dermatosis in which not all the mechanisms involved in its pathogenesis have been elucidated. OBJECTIVE: To study the expression of CS-1 fibronectin, TARC and Th1-associated chemokine receptors in biopsies from allergic patch test reactions. MATERIAL AND METHODS: Thirteen patients already diagnosed with ACD were challenged on the back with the antigen responsible of the disease and macroscopic responses and biopsies taken after 48 h. Skin biopsies from negative control challenge sites, AD and ICD were also taken. Samples were fixed, embedded in paraffin wax and processed in order to perform histological and immunohistochemical studies. RESULTS: All subjects with ACD showed a positive clinical response and a perivascular mononuclear cell infiltration at 48 h, which was not seen in the negative controls. The majority of skin-infiltrating cells were CD4+ and CD8+ and up to 54% or 40% of them expressed CXCR3 or CCR5, respectively. We also showed expression of CS-1 fibronectin in inflamed endothelial cells not only in ACD but also in AC and ICD. In contrast TARC was only expressed in ACD and AC. CONCLUSION: We showed for the first time that CS-1 fibronectin is expressed in dermal vessels from allergic patch tests positive reactions, as well as irritant and atopic skin lesions.

Mesangial nephropathy in Sjögren's syndrome.

We describe a 36-year-old woman with Primary Sjögren's Syndrome (PSS). Purpura, corneal perforation, metabolic acidosis, decreased glomerular filtration, hypokalemia, hyposthenuria, and polyuria were present. Chronic renal insufficiency and renal tubular acidosis type I were diagnosed. Kidney biopsy revealed mesangial glomerulonephritis, interstitial nephritis, and tubular atrophy. Replacement treatment with saliva, tears, and potassium citrate was started. She was given prednisone and cyclophosphamide. This would be the first description of PSS, mesangial glomerulonephritis, and chronic renal insufficiency.