Emergence of visceral leishmaniasis in central Israel.

In 1994-1995, a child and five dogs from villages located between Jerusalem and Tel-Aviv, Israel were diagnosed with visceral leishmaniasis (VL). Based on these findings, the distribution of VL in domestic and wild canids in central Israel was examined. In the two villages where canine index cases were identified, a substantial proportion (11.5%, 14 of 122) of the dogs examined were seropositive. However, the rate of infection in five neighboring villages was only 1% (1 of 99). Parasites were cultured from 92% (12 of 13) of the seropositive dogs biopsied and the strains were characterized as Leishmania infantum by a clamped polymorphic-polymerase chain reaction, monoclonal antibodies, and/or excreted factor serology. The discovery of VL close to major urban centers is an important public health issue. The disease appears to have emerged recently in this area, and it is unclear whether the parasite was re-introduced or was continuously present at low levels in this region. The presence of seropositive wild canids, jackals (7.6%, 4 of 53) and red foxes (5%, 1 of 20), in central Israel, and the reappearance of the jackal population after near extinction suggests that wild canids may play a role in spreading this disease.

Osteogenesis in in vivo diffusion chamber cultures of human marrow cells.

The osteogenic diffusion chamber culture of rodent marrow cells is a well established system. In the present study, marrow cells from children and adult human donors were incubated in diffusion chambers implanted intraperitoneally in athymic mice. After 4 or 8 weeks, the chamber content was examined by light and electron microscopy. Child-cell cultures showed osteogenic tissue consisting of a mineralizing fibrous component and cartilage. Ultrastructurally, the fibrous tissue was similar to osteoid and exhibited osteoblast-like cells and mineralizing nodules. Mineral aggregates were also found in the cartilage. These features in child-cell chambers were similar to those found in control chambers of rabbit marrow cells. Adult-cell chambers showed only unmineralized fibrous tissue. These results render previous findings in animal-cell diffusion chamber systems relevant to the understanding of bone formation in man. It is suggested that the difference between child- and adult-cell chambers reflects an age-related decline in the number of marrow osteoprogenitor cells or their potential to undergo terminal osteogenic differentiation.

Fallopian tubal mucosal damage induced experimentally by Escherichia coli in the rabbit. A scanning electron microscopic study.

Pelvic inflammatory disease is the main cause of infertility due to a tubal factor. The importance of Gramnegative bacteria as major infecting organisms in pelvic inflammatory disease has been recently recognized. The predominant facultative pathogen found in tuboperitoneal fluid from women with salpingitis are coliform bacteria. This study describes an animal model for E. coli mediated tubal mucosa damage. E. coli (10(5)-10(9) bacteria 1 ml) was injected into the right Fallopian tube of sixty rabbits; the left tube injected with culture medium, served as a control. The luminal surface of the oviducts was examined by scanning electron microscope at various intervals after the injection. The degree of damage was found to be dose-dependent and maximal seven days after innoculation. The injury comprised of large areas of deciliation, severely shortened cilia, swollen and adherent throughout their entire length and disappearance of microvilli from the secretory cells. A process of regeneration began two weeks after the innoculation and was completed eight weeks later. Fertility studies carried out after completion of regeneration showed no difference in the ratio of corpora lutea/gestational sacs between the treated and control sides of individual pregnant animals. This model sheds light on the pathogenesis of tubal surface injury by E. coli and suggests that the major effects described are mediated through the endotoxin liberated from the cell wall of these bacteria and that, therefore, antibiotic treatment has no effect on the magnitude of injury. Endosalpingeal regeneration correlates well in this animal model with normal fertility suggesting that morphologic integrity of the surface epithelium may be a good criterion in assessing the reproductive capacity of the fallopian tube.

Mucolipidosis IV: prenatal diagnosis by electron microscopy.

Mucolipidosis IV (ML IV) is a lysosomal storage disease presenting in infancy with cloudy cornea and psychomotor retardation. Our experience with 12 pregnancies at risk for ML IV, monitored by transmission electron microscopy (TEM) studies of cultured amniotic fluid cells, is presented. The prenatal diagnoses were confirmed in the 3 affected and the 8 unaffected pregnancies. In the one pregnancy where no definite diagnosis was reached the pregnancy was terminated. TEM examination of fetal tissues from this pregnancy showed no abnormal lysosomal storage bodies and a review of the cultured amniotic fluid cell sections revealed that the diagnosis of a normal fetus could have been made.

The correction of Hunter fibroblasts by exogenous iduronate sulfate sulfatase: biochemical and ultrastructural studies.

The exogenous addition of iduronate sulfate sulfatase to cultured fibroblasts of Hunter patients resulted in a full correction of the metabolic defect as demonstrated by chemical and ultrastructural analyses. As little as 25% of the normal fibroblasts' enzyme levels were sufficient for this correction. The half-disappearance time of the internalized enzyme was 3-4 days. Prolonged incubation of corrected cells resulted in a gradual reaccumulation of mucopolysaccharides.

Relationship between brain volume and size of dural venous sinuses in human fetuses and adults.

A quantitative study of 50 human fetuses, ranging in size from 64 to 259 mm crown-rump length (CRL), and 12 adult cadavers was carried out. After removal of each brain from the cranial cavity and measurement of its volume (BV), the cross-sectional area (CSA) of the transverse (TS) and sigmoid (SS) dural venous sinuses were measured; the measurements were taken at the same anatomical point in all fetuses and adults. For each fetus and adult, indices CSA . TS/BV and CSA . SS/BV were calculated. The results of the statistical analysis showed that the mean CSA TS/BV . and CSA . SS/BV indices in fetuses were significantly greater than those of the adults, i.e., relative to the size of the brain, the dural venous sinuses in fetuses were greater than in adults. Furthermore, a significant negative correlation was found between the CSA . SS/BV index and the CRL of the fetuses; this means that as the brain and sinuses grow during fetal development, the growth rate of the sinuses decreases. By implication, the rate of growth of the arterial blood supply decreases proportionately. A similar, progressive decrease in the growth rate of the brain takes place during fetal life.

The effects of E. coli endotoxin on the tubal mucosa of the rabbit. A scanning electron microscopic study.

E. Coli endotoxin (5 mg/ml) was injected into the lumen of the right fallopian tube of 25 rabbits. The left tubes, injected with saline, served as controls. The luminal surface of the epithelium of the oviducts was examined by a scanning electron microscope at various intervals after the injection. The damage that was caused by the endotoxin, was found to be maximal by 24 hours after injection. The damage was comprised of adhesions of shortened and swollen cilia and smoothened appearance of the secretory cells due to shortening and swelling of the microvilli. In some areas the epithelium was denuded of cilia. A relative rapid process of repair of the epithelium begins thereafter and by 72 hours, restoration of normal architecture of the mucosa was almost completed. The contralateral control tubes were not affected by the endotoxin. The possible influence of local gram negative bacterial infections, liberating large amounts of endotoxin,on the reproductive capacity is discussed.

A new variant of mannosidosis with increased residual enzymatic activity and mild clinical manifestation.

A partial deficiency of alpha-mannosidase was found in cultured skin fibroblasts, serum, and extracts of leukoytes in two siblings with mild mental retardation, delayed speech, a suggestion of coarse or full facies, and limited mobility of the large joints. All other lysosomal enzymes tested were within the normal range. Their father demonstrated intermediate alpha-mannosidase activity. The addition of 2 mM Zn++ caused a 40% increase of the alpha-mannosidase activity in cell extracts of both patients and control subjects. pH profiles and Cellogel electrophoresis of the patients' cells indicated 20% residual activity of the acidic alpha-mannosidase isoenzyme (pH optimum at 4.0), whereas the activity of the isozyme with pH optimum of 6.0 was normal. Increasing substrate concentration (1--10 mM) demonstrated a 4 to 5-fold increase in the apparent Km of the acidic alpha-mannosidase in the patients' fibroblasts. This residual activity, however, was apparently not sufficient for the normal catabolism of mannose-containing molecules, since electron microscopic examination of the cultured fibroblasts demonstrated numerous lysosomal storage bodies.

Argon laser trabeculotomies in primates: evaluation by histological and perfusion studies.

The continuous argon laser beam has been applied to the trabeculum at the anterior chamber angle of one eye of nine primates. The treatment was evaluated by light and scanning electron microscopy and constant rate infusion studies at different intervals following the laser treatment. This histological studies revealed coagulative necrosis, with trabeculotomies extending into Schlemm's canal in the immediate postoperative period. However, complete closure of the laser-induced trabeculotomies by healing was observed within 1 to 3 weeks following the treatment. Outflow facility data as measured by the perfusion studies revealed somewhat increased values of the treated eye during the early 2 weeks after laser treatment. On longer follow-up, no significant outflow differences were measured between the treated and untreated eyes.

Prenatal diagnosis of mucolipidosis IV by electron microscopy.

Mucolipidosis IV, a recently recognized metabolic storage disease, is characterized clinically by corneal opacity in infancy, full facial features, and psychomotor retardation. Electron microscopy of cells from a 2-year-old affected girl revealed multiple cytoplasmic storage bodies. Cultured amniotic fluid cells, in two subsequent pregnancies, demonstrated similar abnormal storage bodies. Electron microscopic examination of various uncultured tissues from one abortus demonstrated abnormal inclusions in the cells of the brain, cornea, conjunctiva, and other epithelial tissues, thus confirming the prenatal diagnosis. This suggests that mucolipidosis IV is an autosomal recessive trait and demonstrates the efficacy of electron microscopy in the prenatal diagnosis of metabolic storage diseases whose biochemical defect is yet unknown.

Achondrogenesis type I in three sibling fetuses. Scanning and transmission electron microscopic studies.

Three spontaneously aborted fetuses with Type I achondrogenesis in a family with a first cousin marriage are described. Studies by light microscopy revealed abnormal cartilage, enchondral, and periosteal bone, and normal tooth development with abnormal alveolar bone. Electron microscopic studies of cultured skin fibroblasts manifested structurally normal cells. Scanning electron microscopy studies had shown deficient intercartilaginous septa in the metaphysis, with abnormally large calcifying globules. In the diaphysis, the orientation of bone trabeculae and collagen fibers within the trabeculae was disturbed. The numerous osteocytic lucunae were wide and irregular in arrangement and shape. Type 2 achondrogenesis, as studied in these fetuses, is probably a widespread mesenchymal defect, manifested by abnormal calcification and ossification of enchondral and periosteal bone.