Elevated concentration of glycogen in cobalt induced epileptogenic focus

Changes in the concentration of glycogen in various areas of the brain of epileptic rats were investigated. Epilepsy was induced by implantation of cobalt discs on the right sensory motor cortex and epileptic animals have shown clear tonic-clonic jerks of the contra-lateral fore and hind limbs. It was found that glycogen concentration was increased by 29% in the epileptogenic sensory motor cortex as compared to the same area in the contra-lateral hemisphere. Glycogen concentration in other areas within the same hemisphere remained unaffected. Implantation of nickel or copper on the same sensory motor cortex, which did not cause the typical limb jerks of epilepsy, had no effect on glycogen concentration in the same treated areas. Assay of relevant metabolites in the epileptic cortex showed an increase in the concentration of pyruvate and glucose-6-phosphate, by 218% and 112% respectively. The results suggest that the increased glycogen concentration in epileptogenic focus results from increased uptake of glucose due to neuronal hyperexcitability.     

Morbid Obesity Resulting from Inactivation of the Ciliary Protein CEP19 in Humans and Mice

Obesity is a major public health concern, and complementary research strategies have been directed toward the identification of the underlying causative gene mutations that affect the normal pathways and networks that regulate energy balance. Here, we describe an autosomal-recessive morbid-obesity syndrome and identify the disease-causing gene defect. The average body mass index of affected family members was 48.7 (range = 36.7–61.0), and all had features of the metabolic syndrome. Homozygosity mapping localized the disease locus to a region in 3q29; we designated this region the morbid obesity 1 (MO1) locus. Sequence analysis identified a homozygous nonsense mutation in CEP19, the gene encoding the ciliary protein CEP19, in all affected family members. CEP19 is highly conserved in vertebrates and invertebrates, is expressed in multiple tissues, and localizes to the centrosome and primary cilia. Homozygous Cep19-knockout mice were morbidly obese, hyperphagic, glucose intolerant, and insulin resistant. Thus, loss of the ciliary protein CEP19 in humans and mice causes morbid obesity and defines a target for investigating the molecular pathogenesis of this disease and potential treatments for obesity and malnutrition.     

Effect of Stimulation on the Incorporation of 14C from Glial and Neuronal Specific Substrates into Brain Proteins In Vivo and In Vitro

Abstract: The incorporation of amino acids into brain proteins following brachial plexus stimulation (BPS) was studied in anaesthetised Sprague-Dawley rats following injection of radioactive precursors of both neuronal and glial compartments. Following intraperitoneal injection of [¹⁴C]glucose, which is the major neuronal pool precursor, BPS resulted in a significant increase of 379% ( P ± 0.001) in the incorporation of carbon from [¹⁴C]glucose into TCA-insoluble proteins in the contralateral sensorimotor cortex as compared with the ipsilateral area of the same animal. This increase was abolished totally when tetrodotoxin (10 μg ml^-1) was applied topically to the surface of the stimulated area. Following intraperitoneal injection of [¹⁴C]acetate, which is considered to be mainly a glial cell precursor, the same afferent electrical stimuli caused a significant decrease of 21% in the incorporation of amino acids into proteins in the stimulated versus unstimulated sensorimotor cortex. With [4-³H]phenyl-alanine or [l-¹⁴C]leucine as precursors a significant decrease (12%) or no change was recorded, respectively. A similar decrease in protein synthesis in the stimulated sensorimotor cortex was achieved using different routes of injection. No significant changes were observed in the ratio of the specific radioactivities of the total amino acids of the two hemispheres using either precursor. In vitro , synaptosomes showed a large increase in incorporation into proteins after treatment with electrical pulses, both with [¹⁴C]glucose and with [U-¹⁴C]acetate as precursors.     

First Molecular Characterization of Leishmania Species Causing Visceral Leishmaniasis among Children in Yemen

Visceral leishmaniasis (VL) is a debilitating, often fatal disease caused by Leishmania donovani complex; however, it is a neglected tropical disease. L. donovani complex comprises two closely related species, L. donovani that is mostly anthroponotic and L. infantum that is zoonotic. Differentiation between these two species is critical due to the differences in their epidemiology and pathology. However, they cannot be differentiated morphologically, and their speciation using isoenzyme-based methods poses a difficult task and may be unreliable. Molecular characterization is now the most reliable method to differentiate between them and to determine their phylogenetic relationships. The present study aims to characterize Leishmania species isolated from bone marrows of Yemeni pediatric patients using sequence analysis of the ribosomal internal transcribed spacer-1 (ITS1) gene. Out of 41 isolates from Giemsa-stained bone marrow smears, 25 isolates were successfully amplified by nested polymerase chain reaction and sequenced in both directions. Phylogenetic analysis using neighbor joining method placed all study isolates in one cluster with L. donovani complex (99% bootstrap). The analysis of ITS1 for microsatellite repeat numbers identified L. infantum in 11 isolates and L. donovani in 14 isolates. These data suggest the possibility of both anthroponotic and zoonotic transmission of VL-causing Leishmania species in Yemen. Exploring the possible animal reservoir hosts is therefore needed for effective control to be achieved.     

Rehabilitation of a contract killer: caspase-3 directs stem cell differentiation

Activation of caspase-3 is generally acknowledged as a penultimate step in apoptotic cell death pathways. Two studies in this issue of Cell Stem Cell (Fujita et al., 2008; Janzen et al., 2008) provide compelling data to demonstrate that caspase-3 is also a conserved inductive cue for stem cell differentiation.     

Stimulated release of exogenous GABA and glutamate from cerebral cortical synaptosomes and brain slices by bis (acetato) tetrakis (imidazole) copper(II) complex

Severe matermal zinc deficiency has a devastating effect on pregnancy outcome. Studies of humans and experimental animals show that matermal zinc deficiency can cause infertility, prolonged labor, intrauterine growth retardation, teratogenesis, severe immunological deficiencies, or fetal death. The additional need for zinc during pregnancy can be met by an increase in zinc intake. An increase in zinc supplements, when excessive, can cause a decrease in copper. Therefore, it is important to determine the zinc and copper concentrations in embryonic tissue in experimental models and their relationship with embryo number and viability. BALB/c mice were divided into groups according to zinc oral supplementation and gestational age. Phagocytosis was assesed in peritoneal macrophages from dams. The zinc and copper concentrations were obtained by inductively coupled plasma-optical emission spectrometry. Zn and Cu data concentrations in all the analyzed samples were above the detection limits. No spectral interferences were found in both elements (standard reference material was used). Zinc concentrations show a tendency to increase in embryos (14 gestational days and 21 gestational days) supplemented with zinc. Copper concentrations showed a noticeable tendency to diminish (36% and 27%, respectively) in the same period. In contrast, in placenta Zn values were increased by 30% and Cu values were decreased by 26%. We suggest a pivotal role of the placenta metabolism with its homeostatic mechanisms, in these findings. An important increment appeared in the +Zn embryo number (40%) relative to control (−Zn) embryos at 21 d gestational age. Embryo mortality was at 6% in +Zn embryos and at 20% in −Zn embryos. We consider these findings, both in the number and in the viability of +Zn embryos, outstanding.     

The subfamily Encotyllabinae (Monogenea: Capsalidae) with the description of Alloencotyllabe caranxi n. g., n. sp. and Encotyllabe kuwaitensis n. sp.

Alloencotyllabe caranxi n. g., n. sp. is found in groups of 9–15 specimens attached close together to the lower pharyngeal plate of Caranx sp. It is characterized by having an elongate body, a prohaptor with large spines, an armed penis which lies in a pouch and a vaginal pouch guarded by two sets of glands. Encotyllabe kuwaitensis n. sp. is attached individually to the lower pharyngeal plate of Caranx sp. It is characterized by having an elongate body and tandem testes. E. spari is reported from the lower pharyngeal tooth plate of Plectorhynchus cinctus, P. pictus and P. schotaf. All fish hosts were caught in Kuwaiti waters in the Arabian Gulf. The subfamily Encotyllabinae is reviewed and the genus Neoencotyllabe is regarded as a genus inquirendum. The new genus is attached to the subfamily Encotyllabinae.     

Surface morphology of Probolocoryphe uca (Sarkisian, 1957) (Digenea: Microphallidae) from Kuwait Bay

The surface ultrastructure of Probolocoryphe uca (Digenea: Microphallidae), recovered from a rat experimentally fed on crabs, Nanosesarma minutum(Brachyura: Grapsidae), naturally infected with the metacercariae, was studied by scanning electron microscopy. The flukes were leaf-like, ventrally concave and pyriform or ovoid in outline. The anterior end was modified into a sucker-like organ, comprising a protrusible disc-shaped structure surrounded by single-pointed spines. This organ is probably involved in the attachment and feeding process in a manner similar to the action of the oral suckers. Apart from the sucker-like organ, the entire tegumental surface was covered with triangular spines with multi-pointed tips. Ciliated, dome-shaped papillae were observed, singly or in groups, arranged symmetrically on the sucker-like organ and around the oral and ventral suckers. Kuwait Bay constitutes a new geographical record and the crab N. minutum is a new second intermediate host record for P. uca.     

Functional transcriptomics: An experimental basis for understanding the systems biology for cancer cells

We review gene signatures associated with multi-drug resistance and RAS oncogene-mediated transformation as established by microarray analysis and cDNA subtraction. In view of the complexity and diversity of gene expression patterns, we discuss experimental strategies based on signaling pertubation and target gene silencing by RNA interference to narrow down the number of critical genes and to assess their function individually. In addition, we address the question of how oncogenic signaling pathways and the genetic program are wired. DNA methylation of target gene promoters was recognized as one of the critical mechanisms for gene repression, particularly for target genes capable of constraining malignant growth.     

Effect of Depolarizing Agents on the Incorporation of Amino Acids into Soluble Cytoplasmic and Membrane-Bound Proteins of Synaptosome Fractions

Abstract: The incorporation of [U-¹⁴C] protein hydrolysate and [U-¹⁴C]leucine into the trichloroacetic acid (TCA)-insoluble membrane and the soluble synaptoplasm proteins of synaptosomes was studied. Following treatment with the depolarizing agents veratrine, Tityus toxin, or potassium, the specific radioactivity of both precursor pool and proteins was measured to examine the link between protein labeling and the fall in the free amino acid pool due to depolarization-induced release of glutamate and aspartate. By reducing the size of the fall in precursor pool due to depolarization by using a nontransmitter amino acid such as leucine (as compared with the usual use of protein hydrolysate), it was shown that the amount by which the pool is reduced is proportional to the change in the protein labeling observed. These results confirm that membrane depolarization causes a large increase in the labeling of membrane-bound proteins as compared with the soluble synaptosomal proteins.