Decreased aggression and increased repetitive behavior in Pten haploinsufficient mice.

Aggression is an aspect of social behavior that can be elevated in some individuals with autism spectrum disorder (ASD) and a concern for peers and caregivers. Mutations in Phosphatase and tensin homolog (PTEN), one of several ASD risk factors encoding negative regulators of the PI3K-Akt-mTOR pathway, have been reported in individuals with ASD and comorbid macrocephaly. We previously showed that a mouse model of Pten germline haploinsufficiency (Pten(+/-) ) has selective deficits, primarily in social behavior, along with broad overgrowth of the brain. Here, we further examine the social behavior of Pten(+/-) male mice in the resident-intruder test of aggression, using a comprehensive behavioral analysis to obtain an overall picture of the agonistic, non-agonistic and non-social behavior patterns of Pten(+/-) mice during a free interaction with a novel conspecific. Pten(+/-) male mice were involved in less aggression than their wild-type littermates. Pten(+/-) mice also performed less social investigation, including anogenital investigation and approaching and/or attending to the intruder, which is consistent with our previous finding of decreased sociability in the social approach test. In contrast to these decreases in social behaviors, Pten(+/-) mice showed increased digging. In summary, we report decreased aggression and increased repetitive behavior in Pten(+/-) mice, thus extending our characterization of this model of an ASD risk factor that features brain overgrowth and social deficits.

Innovations in oral gene delivery: challenges and potentials.

The oral delivery of peptide, protein, vaccine and nucleic acid-based biotechnology products is the greatest challenge facing the drug delivery industry. Oral delivery is attractive due to factors such as ease of administration, leading to improved patient convenience and compliance, thereby reducing overall healthcare costs. The realization that gene therapy will provide a tangible and potentially huge new therapeutic opportunity has stimulated interest in oral gene delivery. Here we summarize the oral gene delivery vehicles currently in use and highlight potential areas of application, along with the challenges that need to be overcome before this new technology enters the clinic.

labial acts to initiate neuronal fate specification, but not axon pathfinding, in the embryonic brain of Drosophila.

Drosophila embryos lacking the homeotic gene labial (lab) show two types of defects in brain development: (1) cells in the brain lab domain do not express neuronal markers or extend axons, and (2) axons originating from outside the lab domain stop at this region or project ectopically. A severe disruption of neuronal patterning and axon scaffolding is the net result. It is not clear how the absence of Lab can result in both neuronal fate defects and axon pathfinding defects. I have expressed Lab in short pulses in lab loss-of- function embryos, and this gave almost complete rescue; for example, the tritocerebral commissure was restored. Rescue only occurred when Lab was provided at the time when cells in the brain are adopting a neuronal fate. Lab expression later, when the first axons are seen in the lab domain, did not give rescue. I conclude that Lab expression helps to establish neuronal identity in the lab domain, and these neurons act as a permissive substrate for axon extension. However, Lab itself is not required at the time of axon pathfinding through this region.

Mapping studies and genetic analysis of transfer genes of the multi-resistant IncHI2 plasmid, R478.

A bank of transfer-defective Tn7 insertion mutants of the multi-resistant IncHI2 megaplasmid, R478, was generated. Complementation analysis of these mutations identified a large 144-kb transfer-associated region of R478. A 6.8-kb segment from the transfer region was sequenced. The precise locations of Tn7 insertion within four distinct R478::Tn7 transfer-defective mutants were mapped and each insertion was found to disrupt a specific open reading frame. These transfer-associated determinants of R478 were designated htdB (H transfer determinant), htdD, htdT and htdC. Both htdB and htdC encoded amino acid sequences that showed a low homology with pilus biosynthetic proteins encoded by the F plasmid.

Isolation and characterization of hepatocytes and Kupffer cells.

Simplified isolation procedures are described for the parenchymal cell of the liver and the major non-parenchymal cell, the Kupffer cell. Hepatocytes are obtained in a purity of approximately 100%; a yield 10 X 10(6) cells/g liver tissue and the viability is greater than 85%. The recovery of Kupffer cells is 82%, viability 87% and purity 67%. Characterization of Kupffer cells is by the peroxidatic reaction, of hepatocytes by gluconeogenesis and also culture on collagen plates in a non-protein medium yielding albumin.