Improving cancer therapeutics by molecular profiling.

The individualized medicine aims to identify the molecular basis of the individual's response to different therapeutic treatments. Individualized medicine is very relevant for human diseases such as cancer and it has become a major task to accomplish more efficient and specific therapeutics. An individualized response to treatment could underline therapeutic success or failure and, even more, could support the rationale for good or bad prognosis. The use of up to date genomic approaches is changing the way we understand modern medicine in terms of drug efficacy, toxicity and diagnosis. Results from genetic polymorphism studies, gene expression profiling and epigenetics illustrate how pharmacogenomic testing will contribute to the goal of individualized medicine. Antineoplastic drugs are designed to block the anomalous activity of specific molecules (therapeutic targets) that regulate cellular processes such as cell cycle. Understanding the relationship between molecular changes in therapeutic targets and enhanced antitumoral response or chemotherapeutic resistance is crucial to establish the clinical relevance of genomic approaches. The goal of this review is to discuss the basic and the clinical significance of genomic research on drug targets and its impact on the early diagnosis and treatment of cancer. We will also assess how these methodologies could contribute to individualized medicine in oncology. A special focus will be put on oncogenes and tumor suppressor genes. Aspects such as drug efficacy, side effects and the diagnostic value of antineoplastic pharmacogenomic research will be also considered.

Cannabinoid/opioid crosstalk in the central nervous system.

Promising therapeutic uses and a great variety of pharmacological effects are the leading forces that focus actual cannabinoid research. Cannabinoid and opioid systems share neuroanatomical, neurochemical, and paharmacological features. This fact supports the notion that actions induced by each one of these types of drugs involved an interaction between the endogenous opioid and endocannabinoid neuronal systems. Over the last decade our group and others have investigated cannabinoid/opioid crosstalk in the central nervous system by studying the mechanisms underlying pharmacological and biochemical interactions between the two systems in experimental paradigms of antinociception, drug reinforcement, and anxiety. The goal of this review is to revise the latest work done on this subject, with special emphasis on the research done with genetically modified animals. Whereas clinical progress is going ahead slowly, basic research in this area is progressing rapidly. Clinical applications derived from the cannabinoid/opioid crosstalk and based tightly on medical evidence are yet to come, but it is hoped that knowledge of this central messenger interaction will help to develop new alternatives for the treatment of some pathological states.

[Segmental renal infarction presenting as acute flank pain]. / Infarto renal segmentario como causa de dolor agudo en flanco.

OBJECTIVES: To report the clinical case of a male with acute onset right flank pain and significant deterioration of his general status. METHODS: 40-year-old male patient with acute onset of flank pain, progressive worsening, and fever up to 40 degrees C. Blood analysis (hemogram, coagulation tests, and serum ions), urine analysis, and abdominal x-rays, ultrasound and CT scan were performed. After the diagnosis of segmental renal infarction treatment with low molecular weight heparin was started. RESULTS: Patient improved clinically and was discharged from hospital 10 days later and received final discharge as outpatient after 6-month follow-up visit. CONCLUSIONS: Renal infarction should be taken into consideration in the differential diagnosis of acute flank pain, when more frequent causes as renal colic or inflammatory-infectious diseases are ruled out, mainly in patients with cardiovascular diseases because the most frequent cause is embolism.

[Arterial priapism in a prepuberal patient. Conservative treatment]. / Priapismo arterial en paciente prepúber. Tratamiento conservador.

OBJECTIVES: To report the case of a 7-year-old patient with high flow priapism solved with conservative treatment. METHODS: We performed history and physical examination, complementary tests including general blood tests (serum ions, hemogram and coagulation tests), peripheral blood smear, Doppler ultrasound and selective arteriography which led to the diagnosis of high flow priapism. Due to the absence of arteriovenous fistula conservative treatment was chosen. RESULTS: Patient improved clinically after five days, with penile detumescence and spontaneous erections were preserved. CONCLUSIONS: Conservative treatment is a valid option in patients with arterial priapism, mainly in those cases in whom performing embolization of a small fistula seen on arteriography is not possible or when such AV communication is not demonstrated.

Liver portal fibrosis in dioxin receptor-null mice that overexpress the latent transforming growth factor-beta-binding protein-1.

Mice lacking aryl hydrocarbon (dioxin) receptor (AhR) had variable degree of hepatic fibrosis and altered liver architecture. Transforming growth factor-beta (TGF-beta), a major profibrogenic molecule in the liver, is localized to the extracellular matrix by its association to the latent TGF-beta-binding protein-1 (LTBP-1). Very recently, LTBP-1 has been shown to be negatively regulated by the AhR. Embryonic fibroblasts from AhR-null (AhR(-/-)) mice overexpress LTBP-1 and secrete four times more active TGF-beta than wild-type fibroblasts. To test whether TGF-beta and LTBP-1 overexpression colocalize within the fibrotic nodule of AhR(-/-) liver, we have characterized this hepatic portal fibrosis using collagen protein staining, immunohistochemistry and in situ hybridization. LTBP-1 mRNA and protein were overexpressed in the fibrotic region and colocalized with other indicators of fibrosis such as collagen and fibronectin and the fibroblast marker proteins alpha-actin and vimentin. TGF-beta protein also colocalized with fibrosis, although in contrast, TGF-beta mRNA expression, rather than restricted to the fibrotic compartment, was present throughout the hepatic parenchyma and exhibited similar levels in wild-type and AhR(-/-) mice. These results suggest that LTBP-1 targets TGF-beta to specific areas of the liver and that the AhR could be a negative regulator of liver fibrosis, possibly through the control of LTBP-1 and TGF-beta activities.

Repeated administration with Delta9-tetrahydrocannabinol regulates mu-opioid receptor density in the rat brain.

Several studies have demonstrated reciprocal, as well as synergistic interactions between cannabinoid and opioid systems. The aim of this study was to explore the time-related effects of repeated administration of Delta9-tetrahydrocannabinol on mu-opioid receptor autoradiography in various brain regions of the rat. To this aim, the effects of Delta9-tetrahydrocannabinol (Delta9-THC, 5 mg/kg/day; i.p.) were examined after 1, 3, 7 and 14 days of repeated administration on regions containing mu-opioid receptors: (i) forebrain [caudate-putamen, nucleus accumbens (core and shell) and piriform cortex]; (ii) amygdala (medial pars and cortical posteromedial pars), hypothalamus (ventromedial and dorsomedial nuclei, zona incerta), hippocampal regions (CA1, CA2, CA3, dentate girus), hindbrain (substantia nigra and ventral tegmental area); and (iii) thalamus, including 12 thalamic nuclei. In most of these regions, repeated cannabinoid administration increases mu-opioid receptor density; however, the onset, degree of magnitude reached and time-related effects produced by administration with Delta9-tetrahydrocannabinol are dependent upon the brain region examined. It appears that the major increase in mu-opioid receptor density occurs 1 and 3 days after Delta9-THC administration. In some regions, this increase is maintained and, for most of the brain areas examined, this effect is no longer significant by 14 days of administration, suggesting tolerance to cannabinoid treatment. Taken together, the results of this study suggest that cannabinoids produce a time-related differential responsiveness in mu-opioid receptor density in several brain areas that may be relevant to an understanding of the alterations associated with cannabinoid exposure.

Extinction of cocaine self-administration produces alterations in corticotropin releasing factor gene expression in the paraventricular nucleus of the hypothalamus.

The long-term effect of cocaine self-administration on corticotropin releasing factor (CRF) mRNA content in the hypothalamic CRF-containing neurons has not yet been established. The purpose of this study was to examine the time course effects of the extinction of cocaine self-administration behavior on CRF gene expression in the paraventricular nucleus of the hypothalamus (PVN) using in situ hybridization histochemistry (IHHS). Seventy-two littermate male Lewis rats were randomly assigned in triads to one of three conditions: (a) contingent intravenous self-administration of 1 mg/kg/injection of cocaine (CONT), (b) non-contingent injections of either 1 mg/kg/injection of cocaine (NONCONT) or (c) saline yoked (SALINE) to the intake of the self-administering subject. The self-administering rats were trained to self-administer cocaine under a fixed ratio 5 (FR5) schedule of reinforcement for a minimum of 3 weeks. After stable baseline levels of drug intake had been reached, saline was substituted for drug. Following this first extinction period, cocaine self-administration was reinstated for an additional period of 2 weeks. Immediately after cessation of the last session of cocaine self-administration (Day 0) and 1, 5 and 10 days after the second extinction period, animal brains in each triad were removed to be processed for IHHS. CRF mRNA levels in the PVN were significantly lower in the NONCONT cocaine group at Day 0 compared to CONT or SALINE groups. On Day 1, hypothalamic CRF gene expression significantly decreased in the CONT cocaine group with respect to the SALINE group, but there were no differences between the cocaine groups or among the NONCONT cocaine and SALINE groups. After 5 and 10 days of extinction, no differences were found in CRF mRNA content in the PVN between the three conditions of this study. These results suggest that, after the extinction of cocaine self-administration, changes in hypothalamic CRF gene expression are differentially affected depending upon the type of cocaine administration, and that the stages of cocaine withdrawal might not be associated with enduring changes in hypothalamic CRF mRNA levels.

The relative contribution of monoamine oxidase and cytochrome p450 isozymes to the metabolic deamination of the trace amine tryptamine.

Tryptamine is a trace amine in mammalian central nervous system that interacts with the trace amine TA(2) receptor and is now thought to function as a neurotransmitter or neuromodulator. It had been reported that deamination of tryptamine to tryptophol was mediated by CYP2D6, a cytochrome P450 that is expressed in human brain, suggesting that tryptamine may be an endogenous substrate for this polymorphic enzyme. We were unable to confirm this report and have reinvestigated tryptamine metabolism in human liver microsomes (HLM) and in microsomes expressing recombinant human cytochrome P450 and monoamine oxidase (MAO) isozymes. Tryptamine was oxidized to indole-3-acetaldehyde by HLM and recombinant human MAO-A in the absence of NADPH, and indole-3-acetaldehyde was further reduced to tryptophol by aldehyde reductase in HLM in the presence of NADPH. Steady-state kinetic parameters were estimated for each reaction step by HLM and MAO-A. The CYP2D6 substrates bufuralol and debrisoquine showed strong inhibition of both tryptophol production from tryptamine in HLM and the formation of indole-3-acetaldehyde from tryptamine catalyzed by recombinant MAO-A. Anti-CYP2D6 monoclonal antibody did not inhibit these reactions. Pargyline, a nonselective MAO inhibitor, did not show cross inhibition to debrisoquine 4-hydroxylation and dextromethorphan O-demethylation by HLM and recombinant CYP2D6 enzyme. This is the first unequivocal report of the selective conversion of tryptamine to tryptophol by MAO-A. CYP2D6 does not contribute to this reaction.

Gender differences in proenkephalin gene expression response to delta9-tetrahydrocannabinol in the hypothalamus of the rat.

Chronic exposure to delta9-tetrahydrocannabinol (delta9-THC) produces an activation of preproenkephalin (PENK) gene expression in the rat hypothalamus. The levels of circulating gonadal steroids concurrently modulate this neuropeptide in male and female rats. However, whether gonadal steroids regulate delta9-THC effects on PENK gene expression in the hypothalamus of male and female rats remains unknown. To test this hypothesis, experiments were carried out on intact, 2-week-gonadectomized, 1-week-gonadectomized, 1-week-dihydrotestosterone (DHT) replaced male rats, and 2-week-gonadectomized, 1-week-gonadectomized, 1-week-oestradiol replaced female rats. One week after hormonal replacement, animals were treated with vehicle or delta9-THC (5 mg/kg/day, i.p. 7 days). In males, delta9-THC administration to intact animals induced PENK mRNA in the paraventricular nucleus (PVN) and ventromedial nucleus (VMN) of the hypothalamus. Orchidectomy did not affect basal PENK mRNA levels in the PVN, but reduced PENK mRNA levels in the VMN. However, delta9-THC treatment induced PENK gene expression to the same extent in both hypothalamic nuclei of intact, castrated and DHT-replaced males. In females, ovariectomy decreased PENK gene expression in PVN and VMN. delta9-THC administration increased PENK gene expression in castrated females, but had no effect in the oestradiol-replaced group. Taken together, these results suggest gender differences in the response of chronic exposure to cannabinoids on PENK gene expression in the hypothalamus. Furthermore, it appears that alterations in opioid gene expression induced by cannabinoids in female rats depend upon the presence or absence of circulating oestradiol.