21-Gene expression profile assay on core needle biopsies predicts responses to neoadjuvant endocrine therapy in breast cancer patients

This study examined postmenopausal estrogen receptor-positive breast cancer patients who received prospective neoadjuvant endocrine therapy (NAET) with tamoxifen or anastrozole to determine if the 21-gene recurrence score (RS) predicts NAET responses. RS scores were determined from pretreatment core biopsy specimens. Although half of the specimens yielded insufficient RNA, the remaining samples were highly representative. Patients with a low RS tended to respond better than those with an intermediate or high RS (n = 43). Response rates by RS were similar between the tamoxifen and anastrozole groups. Patients with a low RS tended to have better relapse-free survival (RFS) than those with an intermediate or high RS (5y-RFS; 100% vs. 84% and 73%, respectively). These results suggest that RS predicts responses to NAET with tamoxifen or anastrozole. Because this pilot study examined a small sample size, these results should be validated in larger studies.     

Effects of intratumoral injection of CCL2 on monocyte-endothelial cell interactions in mouse pancreatic cancer

OBJECTIVE: Although monocyte infiltration is an important aspect of the host response to tumor growth, the mechanisms of recruitment and their impact on tumor growth are still unknown. The authors studied monocyte-endothelial interaction and the effect of chemokine CCL2 in orthotopic mouse pancreatic cancer. METHODS: The rolling and adhesion of labeled monocytes in peritumoral and intratumoral areas were assessed by using an intravital microscope. Further, the effects of intratumoral injection or superfusion of CCL2 on in situ recruitment of monocytes and other immune cells and adhesion molecules were investigated. RESULTS: More monocytes were recruited in the peritumoral area than in the intratumoral area with increased vascular interaction, and the effect was more apparent by intratumoral CCL2 injection than superfusion. In both CCL2-treated groups infiltration of CD11b(+), CD68(+), and CD4(+) cells were increased, but the magnitude of increase was larger in intratumoral injection. Quantitative RT-PCR for the tumor tissue revealed that ICAM-1 expression was increased by the injection of CCL2. CONCLUSION: These results show intratumoral injection of CCL2 induces effective interaction between monocytes and endothelial cells in the peritumoral area of pancreatic cancer accompanied by the upregulation of ICAM-1 and may possibly become a tool for immunotherapy by promoting the infiltration of immune cells in cancers.     

Configurations of the mitral valve during off-pump coronary artery bypass grafting: endoscopic and three-dimensional analysis

BACKGROUND AND AIM OF THE STUDY: During off-pump coronary artery bypass grafting (CABG), mitral regurgitation (MR) has been experienced in relation to the procedures. The study aim was to evaluate the mitral valve configuration, with particular focus on annular behavior, during off-pump CABG, using cardiac endoscopy and digital 3-D ultrasound sonomicrometry. METHODS: Following implantation of six crystals of the digital 3-D ultrasound sonomicrometer around the mitral annulus, and two crystals on the epicardial base of the papillary muscles, off-pump CABG was simulated in seven beagle dogs. The heart was perfused with pellucid Krebs-Henseleit solution in situ, with controlled left ventricular end-diastolic pressure (LVEDP). The behaviors of the mitral annulus and leaflets were then observed endoscopically with only cardiac displacement, 15 min occlusion of the left anterior descending artery (LAD), or 15 min occlusion of the left circumflex artery (LCx) with cardiac displacement. Dimensions between the crystals were also recorded using digital 3-D ultrasound sonomicrometry. RESULTS: With only cardiac displacement, no MR was observed endoscopically, and no major changes occurred in annular configuration when coronary perfusion was maintained. In one dog, MR was observed only from the anterolateral site after LAD occlusion. MR from the posteromedial site was observed by occlusion of the LCx in all cases, with significant (p < 0.01) enlargement of the mitral annular dimensions. CONCLUSION: Cardiac displacement alone did not cause MR if coronary perfusion was maintained. Occlusion of the LAD rarely caused MR from the anterolateral site, whereas occlusion of the LCx normally caused MR from the posteromedial site; the posteromedial annulus was enlarged, even when the LVEDP was controlled.     

Altered distribution of KCC2 in cortical dysplasia in patients with intractable epilepsy

PURPOSE: To examine the distribution of KCC2, a neuron-specific K(+)-Cl(-) cotransporter, in human cortical dysplasia (CD). METHODS: The immunohistochemical expression of KCC2 was investigated in 18 CD specimens obtained during epilepsy surgery. The histopathologic diagnoses were focal CD (FCD) type I (eight cases), FCD type II (six cases), and hemimegalencephaly (HME; four cases). Tissue sections were immunostained for KCC2 and compared with control sections. RESULTS: In the mature nondysplastic cortex, all the layers showed diffuse neuropil staining for KCC2. The somata were stained much less, although subcortical ectopic neurons displayed dense staining in the cytosol (intrasomatic staining). In FCD type I, the cortex showed neuropil staining for KCC2 with less-stained somata. Aberrant giant pyramidal neurons were also less stained at the soma, whereas immature neurons showed intrasomatic staining. Increased numbers of ectopic neurons with intrasomatic staining were noted in the subcortical white matter. In FCD type II, dysmorphic neurons displayed dense intrasomatic staining with reduced staining of the neighboring neuropils. Balloon cells did not stain for KCC2. Dysmorphic neurons in HME also showed intrasomatic staining. CONCLUSIONS: Neurons in CD tissues expressed KCC2. However, the subcellular distribution of KCC2 was altered, which might have affected the ionic homeostasis of Cl(-) and K(+) involved in epileptic activity within CD tissues.     

A new technique for the mapping of oxygen tension on the brain surface

Most measurements of oxygen tension (PO(2)) in the brain have been performed using oxygen microelectrodes. However, the insertion of microelectrodes into the brain per se causes cortical injury and hence could lead to erroneous PO(2) measurements. The recently developed "quenching lifetime method" requires the injection of fluorescent chemicals into the blood circulation. To address this issue, we tested the feasibility of our O(2)-sensitive fluorescent membrane technique in the rat brain, and visualized the spatial distribution of PO(2) on the brain surface as epifluorescent microscopic patterns. An O(2)-quenching fluorescence dye, tris (1,10-phenanthroline) Ru(2+), was immobilized in a highly gas-permeable, thin silicone-rubber film formed on a microscope coverslip. Unlike the original method, which was intended for transparent rat mesenteric tissue, any change in the redox state in the brain tissue will influence the optical measurement of PO(2). Thus, in the present study, the O(2)-sensing membrane was further coated with a thin opaque silicone-rubber to minimize this type of influence. This new method enabled us to visualize the PO(2) gradient on the rat brain without causing cortical injuries. In an ischemia/reperfusion model using Pulsinelli's four-vessel occlusion rats, the changes in the PO(2) were highly heterogeneous during the ischemic period and this heterogeneity, both temporal and spatial, was higher in the off-arteriolar area than in the peri-arteriolar area.     

A natural scavenger of peroxynitrites, rosmarinic acid, protects against impairment of memory induced by Abeta(25-35)

Peroxynitrite (ONOO⁻)-mediated damage is regarded to be responsible for the cognitive dysfunction induced by amyloid beta protein (Aβ) in Alzheimer's disease (AD). In the present study, we examined the protective effects of rosmarinic acid (RA), a natural scavenger of ONOO⁻, on the memory impairment in a mouse model induced by acute i.c.v. injection of Aβ_25–35. Mice daily received i.p. several doses of RA after the injection of Aβ_25–35. RA prevented the memory impairments induced by Aβ_25–35 in the Y maze test and novel object recognition task. RA, at the effective lowest dose (0.25 mg/kg), prevented Aβ_25–35-induced nitration of proteins, an indirect indicator of ONOO⁻ damage, in the hippocampus. At this dose, RA also prevented nitration of proteins and impairment of recognition memory induced by ONOO⁻-i.c.v.-injection. Co-injection of the non-memory-impairing dose of ONOO⁻ with Aβ_25–35 blocked the protective effects of RA (0.25 mg/kg). These results demonstrated that the memory protective effects of RA in the neurotoxicity of Aβ_25–35 is due to its scavenging of ONOO⁻, and that daily consumption of RA may protect against memory impairments observed in AD.     

Effects of a new orally active dopamine prodrug, docarpamine, on refractory ascites: a pilot study

OBJECTIVE: Refractory ascites is a debilitating condition in patients with cirrhosis. Recently, docarpamine, an orally active dopamine prodrug, was reported to increase renal blood flow, glomerular filtration, and sodium excretion. This suggests docarpamine may be useful for the treatment of refractory ascites. METHODS: In this study, we investigated docarpamine metabolism in cirrhotic patients and its effect on refractory ascites. RESULTS: Blood samples were obtained from seven cirrhotic patients and six healthy subjects after administration of 750 mg docarpamine, and plasma levels of free dopamine were measured. In healthy subjects, maximum plasma concentration (Cmax), time taken to reach Cmax (Tmax), elimination half-life (T(1/2)), and area under the plasma concentration-time curve (AUC) of plasma free dopamine were 76.8 +/- 24.1 ng/ml, 1.3 +/- 0.2 h, 0.8 +/- 0.1 h, and 97.5 +/- 21.1 ng x h/ml, respectively. In patients with cirrhosis, Cmax (53.1 +/- 24.9 ng/ml), T(1/2) (0.8 +/- 0.1 h), and AUC (100.6 +/- 45.6 ng x h/ml) were no different from healthy subjects when comparing each parameter, whereas Tmax (2.7 +/- 0.2) was significantly longer than that of healthy subjects. We treated 10 cirrhotic patients with refractory ascites with docarpamine or placebo and the same dose of diuretics used before hospitalization. After 8 wk of docarpamine treatment, ascites disappeared completely in three of the five patients and decreased in the remainder. However, in five patients treated with placebo, ascites was not changed or increased. Side effects were not observed in any case. CONCLUSIONS: Docarpamine was found to metabolize in cirrhotic patients as well as in normal subjects and may be an effective treatment for refractory ascites.     

Drug Discovery for Overcoming Chronic Kidney Disease (CKD): New Therapy for CKD by a (Pro)Renin-Receptor–Blocking Decoy Peptide

Discovery of the (pro)renin receptor uncovered a novel function of renin/prorenin as the receptor ligands in addition to the enzyme and its precursor. The bindings of renin and prorenin to the (pro)renin receptor trigger two major pathways: the angiotensin II–dependent pathway as a result of the enzymatic activation of renin/prorenin and the angiotensin II– independent intracellular pathway involving hypertrophic, hyperplastic, and profibrotic signals. A specific blocker of the receptor was discovered through identification of the amino acid sequence of prorenin prosegment that binds to the receptor and leads to non-proteolytic conversion of prorenin to its active form. A peptide containing this sequence was found to block the binding of prorenin to its receptor. Its infusion in animal models of diabetes and low-renin hypertension significantly inhibited the development and progression of nephropathy, but (pro)renin receptor blockade had no benefit in the clipped kidney of 2K1C rats or rat models of high-renin hypertension. Since renin is still active without a (pro)renin receptor, (pro)renin-receptor blockade elicits a maximum benefit under low-renin conditions. Thus, (pro)renin-receptor blockade can be a useful therapy for chronic kidney disease with low renin levels in the plasma.     

Long-term effects of neonatal MK-801 treatment on prepulse inhibition in young adult rats

Rationale

Blockade of N-methyl-d-asparate (NMDA) receptors has been shown to produce some of the abnormal behaviors related to symptoms of schizophrenia in rodents and human. Neonatal treatment of rats with non-competitive NMDA antagonists has been shown to induce behavioral abnormality in a later period.

Objectives

The aim of this study was to determine whether brief disruption of NMDA receptor function during a critical stage of development is sufficient to produce sensorimotor-gating deficits in the late adolescence or early adulthood in the rat.

Methods

Male pups received the NMDA receptor blocker MK-801 (0.13 or 0.20 mg/kg), or an equal volume of saline on postnatal day (PD) 7 through 10. The animals were tested twice for prepulse inhibition (PPI) and locomotor activity in pre- (PD 35-38) and post- (PD 56-59) puberty.

Results

Neonatal exposure to both doses MK-801 disrupted PPI in the adolescence and early adulthood. Low-dose MK-801 elicited long-term effects on startle amplitudes, whereas high-dose MK-801 did not. Neither dose of MK-801 showed a significant effect on spontaneous locomotor activity, whereas the high dose attenuated rearing.

Conclusions

The results of this study suggest neonatal exposure to MK-801 disrupted sensorimotor gating in the adolescence and early adulthood stages. These findings indicate that rats transiently exposed to NMDA blockers in neonatal periods are useful for the study of the pathophysiology and treatment of schizophrenia.