Synthesis and biological evaluation in vitro and in mammalian cells of new heteroaryl carboxyamides as HIV-protease inhibitors.

New heteroaryl HIV-protease inhibitors bearing a carboxyamide spacer were synthesized in few steps and high yield, from commercially available homochiral epoxides. Different substitution patterns were introduced onto a given isopropanoyl-sulfonamide core modifying the type of heteroarene and the central core, with the presence of either H or benzyl group. Their in vitro inhibition activity against recombinant protease showed a general beneficial effect of carboxyamide moiety, the IC50 values ranging between 1 and 15nM. In particular benzofuryl derivatives showed IC50 values among the best for such structurally simple inhibitors. Docking analysis allowed to identify the favorable situation of such benzofuryl derivatives in terms of number of interactions in the active site, supporting the experimental results on activity. The inhibition activity of such molecules has been also evaluated in HEK293 cells expressing the protease fused to green fluorescent protein, by western blotting analysis, fluorescence microscopy and cytofluorimetry.

The hepatitis B x antigen anti-apoptotic effector URG7 is localized to the endoplasmic reticulum membrane.

Hepatitis B x antigen up-regulates the liver expression of URG7 that contributes to sustain chronic virus infection and to increase the risk for hepatocellular carcinoma by its anti-apoptotic activity. We have investigated the subcellular localization of URG7 expressed in HepG2 cells and determined its membrane topology by glycosylation mapping in vitro. The results demonstrate that URG7 is N-glycosylated and located to the endoplasmic reticulum membrane with an Nlumen-Ccytosol orientation. The results imply that the anti-apoptotic effect of URG7 could arise from the C-terminal cytosolic tail binding a pro-apoptotic signaling factor and retaining it to the endoplasmic reticulum membrane.

A neuroprotective phase precedes striatal degeneration upon nucleolar stress.

The nucleolus is implicated in sensing and responding to cellular stress by stabilizing p53. The pro-apoptotic effect of p53 is associated with several neurodegenerative disorders, including Huntington's disease (HD), which is characterized by the progressive loss of medium spiny neurons (MSNs) in the striatum. Here we show that disruption of nucleolar integrity and function causes nucleolar stress and is an early event in MSNs of R6/2 mice, a transgenic model of HD. Targeted perturbation of nucleolar function in MSNs by conditional knockout of the RNA polymerase I-specific transcription initiation factor IA (TIF-IA) leads to late progressive striatal degeneration, HD-like motor abnormalities and molecular signatures. Significantly, p53 prolongs neuronal survival in TIF-IA-deficient MSNs by transient upregulation of phosphatase and tensin homolog deleted on chromosome 10 (PTEN), a tumor suppressor that inhibits mammalian target of rapamycin signaling and induces autophagy. The results emphasize the initial role of nucleolar stress in neurodegeneration and uncover a p53/PTEN-dependent neuroprotective response.

In vitro properties of small ruminant lentivirus genotype E.

Small ruminant lentivirus genotype E lacks the dUTPase subunit and vpr-like gene. Two strains (Roccaverano and Seui) with identical genetic organization have been described, with the env HV1-HV2 domains being the most divergent. Although dUTPase and vpr-like deletions have been involved in the RT fidelity in non dividing cells, both strains were able to replicate efficiently in blood derived macrophages (BDM), while virus production of E1 subtype was reduced or abrogated in replicating fibroblastic-like cells. The transcriptional activity of genotype E was similar in these two cellular populations. When viral pseudotypes were generated with the env of both viruses, Roccaverano pseudotype displayed a paranuclear localization on BDM, suggesting a different mechanism of entry. Polymorphic GAS and TAS sites in the U3 region, further suggest that a population different from classically activated macrophages can be infected by these viruses, opening new insights into lentiviruses with low or null pathogenic potential.

Adult neurogenesis without germinal layers: the "atypical" cerebellum of rabbits.

Unlike non mammalian vertebrates, adult neurogenesis in mammals is detectable in highly restricted brain sites. Persistent neurogenesis is thought to depend on stem cells residing in neural stem cell niches which are remnants of the embryonic germinal layers. Local progenitors which retain some proliferative capacity have been identified in the mature brain parenchyma, yet they do not support a constitutive, 'actual' neurogenesis, but rather a 'potential' neurogenesis which does not extrinsecate fully and spontaneously in vivo. In contrast with such a view, genesis of neuronal and glial cells from local progenitors does occur in the peripuberal and adult rabbit cerebellum. This process is independent from persisting germinal layers and involves different cell populations.

American Academy of Pediatrics. Committee on Substance Abuse. Marijuana: A continuing concern for pediatricians.

Marijuana, the common name for products derived from the plant Cannabis sativa, is the most common illicit drug used by children and adolescents in the United States.(1) Despite growing concerns by the medical profession about the physical and psychological effects of its active ingredient, Delta-9-tetrahydrocannabinol, survey data continue to show that increasing numbers of young people are using the drug as they become less concerned about its dangers.(1)

Assessment, diagnosis, and treatment of the dually diagnosed adolescent.

The awareness of the prevalence and presentation of psychiatric diagnoses is essential in the quality treatment of adolescent substance abusers. An ongoing relationship with a psychiatrist who can be available for consultation as needed is helpful. Careful observation and history-taking and appropriate consultation results in better detection and treatment of comorbid disorders and, ultimately, in better treatment of the patient.

High rate of affective disorders in probands with attention deficit disorder and in their relatives: a controlled family study.

In a controlled family study of attention deficit disorder, data were collected on first-degree relatives of 22 children with attention deficit disorder and 20 normal children. The rate of major affective disorder was significantly higher in the attention deficit disorder probands (32%) and their relatives (27%) than in the normal control subjects (0%) and their relatives (6%). The findings indicate that attention deficit disorder is associated with higher risk for affective disorder and suggest that probands who have both disorders may represent a distinct subgroup.

Disabling obsessive thoughts responsive to antidepressants.

The authors report four patients with disabling obsessive thoughts who responded in dramatic fashion to antidepressant medication. None met criteria for major depression. This response is discussed in light of the current literature on obsessive-compulsive disorder.

A family study of patients with attention deficit disorder and normal controls.

In a family study of Attention Deficit Disorder (ADD), we collected data on first-degree relatives of 22 children with ADD and 20 normal children. The morbidity risk for ADD was 31.5% in the first group. This was significantly higher than the rate of 5.7% in the control group. Relatives of ADD probands were also shown to be at higher risk for Oppositional Disorders and Major Depressive Disorder (MDD). The findings indicate that ADD is a familial disorder associated with increased familial risk of other psychiatric disorders.