Hippocampal subfields revealed through unfolding and unsupervised clustering of laminar and morphological features in 3D BigBrain.

The internal structure of the human hippocampus is challenging to map using histology or neuroimaging due to its complex archicortical folding. Here, we aimed to overcome this challenge using a unique combination of three methods. First, we leveraged a histological dataset with unprecedented 3D coverage, BigBrain. Second, we imposed a computational unfolding framework that respects the topological continuity of hippocampal subfields, which are traditionally defined by laminar composition. Third, we adapted neocortical parcellation techniques to map the hippocampus with respect to not only laminar but also morphological features. Unsupervised clustering of these features revealed subdivisions that closely resemble gold standard manual subfield segmentations. Critically, we also show that morphological features alone are sufficient to derive most hippocampal subfield boundaries. Moreover, some features showed differences within subfields along the hippocampal longitudinal axis. Our findings highlight new characteristics of internal hippocampal structure, and offer new avenues for its characterization with in-vivo neuroimaging.

Metallothionein in human gingival amalgam tattoos.

Amalgam tattoos occur when small particles of dental amalgam, composed largely of silver (Ag) and mercury (Hg), are inadvertently implanted into oral soft tissues during dental procedures. Metallothioneins (MTs) are ubiquitous, low molecular weight, cysteine-rich, metal-binding proteins that are inducible by many agents including metals and may be involved in the detoxification of toxic metals such as Hg. In this study, the correlation between MT expression and amalgam tattoos in human gingiva was investigated using energy-dispersive X-ray microanalysis (EDX) and immunohistochemical techniques. Light microscopically, amalgam tattoos presented as either fine granular particles or larger discrete opaque globular particles in connective tissues. EDX revealed the smaller particles to be silver sulphide (Ag(2)S), while the larger particles exhibited a shell of Ag(2)S that contained irregularly distributed masses of Ag and Hg. Particles of tin (Sn) were also found. No MT staining was observed in collagen, fibroblasts or blood vessels in areas exhibiting abundant amounts of embedded fine granular Ag(2)S particles. Blood vessels exhibiting relatively few amalgam particles stained positively for MT. Cells with the morphological features of histiocytes located directly adjacent to larger pieces of amalgam showed intense MT staining. These results indicate that amalgam tattoos contain no Hg or free Ag except in large globular pieces of amalgam, which still contain Hg and which induce MT expression in adjacent histiocytes. This suggests that Hg leaching from impacted dental amalgam particles induces MT, while residual Ag(2)S and Sn particles do not. MT may therefore act to reduce Hg exposure in patients with amalgam tattoos.

Effect of age and sex on liver damage due to excess dietary copper in Fischer 344 rats.

OBJECTIVE: The aim of this study was to investigate the morphologic and biochemical effects of excess dietary copper in young and adult rats of different sex. METHODS: Adult Fischer 344 male and female rats were given a diet containing 1500 ppm copper for 18 weeks. Young male and female rats were fed a similar copper-loaded diet from birth until 16 weeks of age. Age- and sex-matched control rats were fed a normal rodent diet (<10 ppm copper). Serum liver enzyme activity was determined in all rats. Livers were sampled for histology, histochemistry (rhodanine), immunohistochemical detection of metallothionein and copper analysis by atomic absorption spectrophotometry. Hepatic metallothionein and zinc concentrations were measured in adult rats. RESULTS: Excess dietary copper caused substantial liver injury, as evidenced by morphologic changes and increased activity of serum ALT, GGT, and SDH. All copper-loaded rats had significantly (p< 0.05) increased hepatic copper concentrations compared to controls. However, young copper-loaded rats accumulated more hepatic copper, had more severe liver changes, and had higher serum liver enzyme activities than adult rats. Histologic changes in copper-loaded rats consisted of multifocal hepatitis and widespread single-cell necrosis. Cytoplasmic copper was detected histochemically in centroacinar zone 1 (portal) and mid-zone in copper-loaded rats. Immunoreactivity for metallothionein was prominent in necrotic hepatocytes and within inflammatory foci in copper-loaded rats. However, differences in hepatic metallothionein concentrations were not detected between adult copper-loaded and control rats. CONCLUSIONS: Young Fischer 344 rats are more susceptible than adults to copper-induced liver injury.

Role of placental metallothionein in maternal to fetal transfer of cadmium in genetically altered mice.

The role of placental metallothionein (MT) as a barrier for maternal to fetal transfer of cadmium (Cd) was investigated using mice which overexpressed the MT-1 isoform (MT-1*), mice which did not express the MT-1 and 2 isoforms (MT-null) and control mice (C57BL/6). In addition, immunohistochemical localization of MT in the placenta was determined in these mice. Two days prior to parturition, the mice were injected with radioactive 109Cd chloride (4 microCi, 0.6 ng Cd/mouse) and killed 24 h later. Organs and fetuses were collected and radioactivity, MT and metal levels were measured. Cd accumulated mainly in the liver and kidney (80% of administered dose) with very low levels (0.1-0.3%) detected in fetuses. When analyzed on a per organ or per gram basis, MT-null fetuses accumulated significantly more Cd (3-10-fold) than the control fetuses and there was no significant difference in fetal Cd accumulation in the MT-1* and control fetuses. As expected, MT and zinc levels were higher in MT-1* than C57BL/6 mice and no MT was detected in MT-null mice. Most striking was the high hepatic MT levels in MT-1* dams (4 mg/g). Immunohistochemical analysis showed MT staining in spongiotrophoblasts, glycogen cells, visceral yolk sac, trophoblast giant cells and maternal decidual cells with the MT-1* placenta staining much more intensely as compared to control placenta. The results suggest that placental MT reduces maternal to fetal Cd transfer, however the low doses of Cd administered in the present experiment resulted in high levels of Cd accumulation in liver and kidney in all groups of mice with a low concentration of Cd reaching the placenta. Thus, the role of placental MT as a barrier for Cd is inconclusive.

Developmental changes in hepatic metallothionein, zinc, and copper levels in genetically altered mice.

Using mice that either overexpress metallothionein 1 (MT-1*) or do not express metallothionein 1 and 2 (MT-null) and a control strain (C57BL/6), the essential metal storage function of hepatic metallothionein and its subcellular localization were investigated during development. Hepatic metallothionein, zinc, and copper levels were measured in all groups from gestational day 20 to 60 days of age. Hepatic metallothionein levels were maximal during the perinatal period in both MT-1* and C57BL/6 mice with levels approximately three times higher in MT-1* mice. MT-null mice had no detectable hepatic metallothionein throughout development. Hepatic zinc levels were highest in the neonatal period of MT-1* and C57BL/6 mice and declined to adult levels by 30 days of age, while hepatic zinc levels in MT-null mice did not vary markedly throughout development. Hepatic copper profiles were very similar in MT-1* and MT-null mice as compared with the C57BL/6 mice. Correlation analysis showed a strong positive correlation between hepatic metallothionein and zinc levels in MT-1* mice, moderate correlation between hepatic metallothionein and metals in C57BL/6 mice, but only a very weak correlation between hepatic metallothionein and copper levels in MT-1* mice. Immunohistochemical localization showed specific nuclear staining in both MT-1* and C57BL/6 mice during the neonatal period with a gradual shift to the cytoplasm. The results show that hepatic metallothionein is a major determinant of zinc but not copper levels during murine development. Additionally, hepatic metallothionein levels and localization are regulated in a similar manner in MT-1* and C57BL/6 mice. The MT-null mice maintain a basel level of zinc sufficient for development, which was found to be 15.9 micrograms/g. This value was similar to the levels of hepatic zinc that was not bound to metallothionein in MT-1* and C57BL/6 mice during development.

The T-cell independent antigen, NP-ficoll, primes for a high affinity IgM anti-NP response.

In a number of different strains of inbred mice, immunization with a hapten coupled to a protein carrier results in production of homogeneous serum antibodies. At the genetic level this corresponds to the use of a very limited set of variable region genes in the actively secreting B-cells. In contrast, immunization with the same hapten coupled to a T-cell independent (TI) carrier produces a heterogeneous antibody response. Here we show that successive immunizations of C57BL/6 mice, first with the hapten NP coupled to ficoll, a TI carrier, and then one month later with a subliminal dose of the same hapten coupled to a protein carrier, generate a novel set of hybridomas. These hybridomas produce antibodies which are of the IgM isotope and which lack somatic mutation. Some of these antibodies have a much higher affinity for NP than do antibodies which use the prototypical gene combination (VH186.2-lamda 1) of the strain specific response in C57BL/6 mice.

Intravenous prochlorperazine for the rapid control of nausea and vomiting in acute myocardial infarction: a clinical observation.

Sixteen patients with acute myocardial infarctions who were either vomiting or nauseated were given an intravenous injection of prochlorperazine. All patients obtained relief with exception of one patient who was in acute renal failure. No patient developed symptomatic hypotension. Intravenous prochlorperazine in the dose of 2.5 mg is a rapid, effective, and safe method to relieve vomiting and nausea in patients who have sustained an acute myocardial infarction.