Id-1 is a key transcriptional regulator of glioblastoma aggressiveness and a novel therapeutic target.

Glioblastoma is the most common form of primary adult brain tumors. A majority of glioblastomas grow invasively into distant brain tissue, leading to tumor recurrence, which is ultimately incurable. It is, therefore, essential to discover master regulators that control glioblastoma invasiveness and target them therapeutically. We show here that the transcriptional regulator Id-1 plays a critical role in modulating the invasiveness of glioblastoma cell lines and primary glioblastoma cells. Id-1 expression levels positively correlate with glioma cell invasiveness in culture and with histopathologic grades in patient biopsies. Id-1 knockdown dramatically reduces glioblastoma cell invasion that is accompanied by profound morphologic changes and robust reduction in expression levels of "mesenchymal" markers, as well as inhibition of self-renewal potential and downregulation of glioma stem cell markers. Importantly, genetic knockdown of Id-1 leads to a significant increase in survival in an orthotopic model of human glioblastoma. Furthermore, we show that a nontoxic compound, cannabidiol, significantly downregulates Id-1 gene expression and associated glioma cell invasiveness and self-renewal. In addition, cannabidiol significantly inhibits the invasion of glioblastoma cells through an organotypic brain slice and glioma progression in vivo. Our results suggest that Id-1 regulates multiple tumor-promoting pathways in glioblastoma and that drugs targeting Id-1 represent a novel and promising strategy for improving the therapy and outcome of patients with glioblastoma.

Cannabidiol enhances the inhibitory effects of delta9-tetrahydrocannabinol on human glioblastoma cell proliferation and survival.

The cannabinoid 1 (CB(1)) and cannabinoid 2 (CB(2)) receptor agonist Delta(9)-tetrahydrocannabinol (THC) has been shown to be a broad-range inhibitor of cancer in culture and in vivo, and is currently being used in a clinical trial for the treatment of glioblastoma. It has been suggested that other plant-derived cannabinoids, which do not interact efficiently with CB(1) and CB(2) receptors, can modulate the actions of Delta(9)-THC. There are conflicting reports, however, as to what extent other cannabinoids can modulate Delta(9)-THC activity, and most importantly, it is not clear whether other cannabinoid compounds can either potentiate or inhibit the actions of Delta(9)-THC. We therefore tested cannabidiol, the second most abundant plant-derived cannabinoid, in combination with Delta(9)-THC. In the U251 and SF126 glioblastoma cell lines, Delta(9)-THC and cannabidiol acted synergistically to inhibit cell proliferation. The treatment of glioblastoma cells with both compounds led to significant modulations of the cell cycle and induction of reactive oxygen species and apoptosis as well as specific modulations of extracellular signal-regulated kinase and caspase activities. These specific changes were not observed with either compound individually, indicating that the signal transduction pathways affected by the combination treatment were unique. Our results suggest that the addition of cannabidiol to Delta(9)-THC may improve the overall effectiveness of Delta(9)-THC in the treatment of glioblastoma in cancer patients.

Radiation-induced caspase-8 mediates p53-independent apoptosis in glioma cells.

Malignant gliomas are almost uniformly fatal and display exquisite radiation resistance. Glioma cells lacking wild-type (WT) p53 function are more susceptible to radiation-induced apoptosis than their isogenic counterparts expressing WT p53. We explored the mechanisms of such apoptosis and found that, in the absence of WT p53, radiation increases caspase-8 expression and activity. Inhibition of caspase-8 expression using caspase-8 antisense or small interfering RNA (siRNA) oligonucleotides partially blocks radiation-induced apoptosis. In contrast, inhibition of the mitochondrial death pathway by expression of Bcl-2 has no effect on radiation-induced caspase-8 activity or apoptosis. Our data indicate that, in contrast to commonly accepted models of p53-dependent radiation-induced apoptosis, in our cell system, radiation relies on caspase-8 activity to help mediate p53-independent cell death. In a system of inducible E2F1 activity, E2F1 activated caspase-8 and, accordingly, decreased cellular viability, effects that were abolished by caspase-8 siRNA. In this model, in the absence of WT p53, p21Cip1 is not induced, and E2F1 activity is sustained and allows transcription and activation of caspase-8. This model may explain why p53 mutations in adult gliomas paradoxically correlate with improved survival and enhanced response to radiation.

Bioenergy definitions and research guidelines.

A model for the functional and observable interrelation among the various components in a physical bioenergy system is presented. The analogy is made between electric circuits and electromagnetic interactions, and contact and noncontact bioenergy transfer. It is postulated that there exists some form of bioenergy that has the capacity to do work and that this energy behaves in a manner similar to electricity in that the physical concepts of electromotive force, current, and impedance have their equivalents in bioenergy. It is further postulated that these analogous components are related by an equivalent to Ohm's and other physical laws of electricity. This is extended to a conjecture that bioenergy healing is the transfer of information from a practitioner to a healee. Research guidelines for bioenergy measurements are presented, including basic measurement practices for electrical and electromagnetic systems through direct measurements and the use of indirect measurement experiments for detecting these or other forms of bioenergy transfer. The research guidelines are divided into 2 sections: those involving direct measurement of the physical electrical properties of a practitioner, in particular the difficulties associated with electrical measurements of extremely low-level signals outside of a Faraday shield or electromagnetic measurements outside of a radio frequency anechoic chamber; and those for conducting experiments in which the effects of bioenergy are being investigated on the healee or other target system without direct measurements of the means for bioenergy transfer.

Distant healing intention: definitions and evolving guidelines for laboratory studies.

This paper provides definitions and a discussion of evolving guidelines for conducting research on the effects of distant healing intention (DHI) on living systems in the laboratory. We consider the relevance of DHI laboratory research to applied healing, special theoretical challenges, and other considerations that distinguish DHI research from other domains of laboratory science. Two sample protocols for investigating DHI are provided, one involving the human autonomic nervous system as the "target" of distant intention, and the other involving cell cultures. In essence, DHI differs from other alternative healing modalities because it postulates that mental intention alone can affect living systems at a distance, unbounded by the usual constraints of both space and time. This postulate challenges scientific assumptions that often go unexamined, including the nature of causality, the distinction between subjective and objective states, and the efficacy of double-blind protocols in controlling for experimenters' intentions. Previous laboratory research in this domain suggests that DHI effects warrant serious study, but most scientists and funding agencies are unaware of the evidence or the relevant literature. By following these evolving guidelines, researchers' designs and their ultimate publications will conform more closely to the quality standards expected by scientific journals, and such publications will in turn attract the attention of a broader range of scientists. This seems especially important for alternative healing research in general and for distant healing in particular; both realms enjoy broad public support but have largely eluded serious attention by mainstream science.