Background.-The pathophysiology of chronic migraine (CM) is not fully understood. We aimed to examine transcranial magnetic stimulation (TMS) indices of cortical excitability in patients with CM and also performed PET studies to ascertain if there were any areas of activation and inhibition for possible correlation.Methods.-Excitability of the cortex was assessed by a reliable parameter of magnetic suppression of perceptual accuracy (MSPA) profiles using transcranial magnetic stimulation in 25 patients with CM. Of these 10 patients were also studied with ( 18 F-FDG PET) scans. Results.-MSPA demonstrated decreased inhibition in CM compared to normal controls and episodic migraine. The percentage of letters reported correct at 100 ms was 84.37 for CM compared to 19.14 for normal controls and 57.41 for episodic migraine. The PET evaluation in 10 subjects demonstrated increased cerebral metabolism in areas of in the brainstem compared to the global flow. There were also decreased areas of cerebral metabolism in the medial frontal and parietal as well as the somatosensory cortex.Conclusions.-Patients with CM appear to be characterized by reduced visual suppression correlating with high cortical excitability. In a cohort of these subjects there was brainstem activation and inhibition in certain areas of the cortex suggesting a potential dysfunction in the inhibitory pathways.

Osteopontin is a secreted glycosylated phosphoprotein known to be involved in numerous physiologic functions and associated with the late stages of various cancers. We used preneoplastic and neoplastic mouse models of prostate cancer to determine the onset of elevated expression of osteopontin in the development of this disease. Osteopontin alterations occurred early in the disease with dysregulated expression observed in lesions of low-grade prostatic intraepithelial neoplasia (PIN). Over time, osteopontin expressing dysplastic cells seemed to increase in number in high-grade PIN and increased further in adenocarcinoma, and in metastasis, almost all of the cancer cells immunohistochemically stained positive for osteopontin overexpression. We examined the biological properties of human prostate cancer cell lines LNCaP and PC-3, in which osteopontin overexpression was achieved via lentiviral gene transduction. Evidence was obtained that osteopontin could contribute to a proliferative advantage in both cell types, although more significantly in LNCaP than PC-3. Osteopontin also influenced their in vitro invasive ability, and again, most strikingly in the weakly oncogenic LNCaP. Furthermore, excess osteopontin induced the LNCaP cells to acquire a strong intravasation potential in vivo in the chicken embryo chorioallantoic membrane assay for blood vessel penetration. These results establish a correlation between an increased gradient of osteopontin expression throughout the stages of murine prostate cancer, beginning from the preneoplastic lesions to distant metastases that suggests a proliferative and invasive advantages to those prostate tumor cells overexpressing osteopontin. Together, these findings support a strategy designed to target osteopontin in the context of prostate cancer therapy. (Cancer Res 2006; 66(2): 883-8)

Objective To assess the risk of hypertension in patients with migraine who received erenumab in clinical trials and in the postmarketing setting. Background Erenumab is a monoclonal antibody for migraine prevention that targets the calcitonin gene‐related peptide (CGRP) receptor. Hypertension is a theoretical risk for inhibitors of the CGRP pathway. Although no evidence of an association between erenumab treatment and hypertension was observed during the clinical development program, adverse events (AEs) of hypertension have been identified in the postmarketing setting. Methods Safety data from four phase 2 and phase 3 clinical trials were used to perform a pooled analysis of hypertension AEs in patients with migraine receiving erenumab. Postmarketing AEs of hypertension were identified from the Amgen Global Safety database from May 17, 2018, through January 31, 2020. Results In the pooled analysis of clinical trials, hypertension AEs (placebo, 9/1043 [0.9%]; erenumab 70 mg, 7/893 [0.8%]; erenumab 140 mg, 1/507 [0.2%]) and percentage of patients initiating medication to treat hypertension (12/1043 [1.2%], 7/893 [0.8%], 1/507 [0.2%], respectively) were similar across treatment groups. A total of 362 AEs of hypertension were identified from the postmarketing setting, 26.2% (95/362) of which were serious, >245,000 patient‐years of exposure. The exposure‐adjusted incidence of hypertension was 0.144 per 100 patient‐years. Conclusions Clinical trials did not demonstrate an increased risk of hypertension with erenumab compared with placebo, and AE rates of hypertension reported with erenumab in the postmarketing setting were generally low. Additional data are needed to fully characterize the extent to which hypertension is a risk associated with erenumab.

Objective: To assess cardiovascular (CV) safety of erenumab in clinical trial patients associated with degree of CV risk.Background: Hypertension has been considered a theoretical risk associated with the inhibition of the calcitonin gene-related peptide pathway in migraine management, particularly in a patient population with pre-existing CV risk factors.Methods: Data pooled from four double-blind, randomized trials were used to assess blood pressure (BP) changes and CV safety in patients grouped based on 10-year risk of cardiac, cerebrovascular, and peripheral artery disease as no-risk-factors, low-risk (>0% to ≤10%), moderate-risk (>10% to ≤20%), and high-risk (>20%) categories. CV safety was assessed as ischemic cardiovascular and cerebrovascular adverse events (ICCAE). Results: There was no apparent difference between placebo-(N = 1032) and erenumabtreatment groups (70 mg, N = 885; 140 mg, N = 504) in clinical worsening of BP category from baseline to Months 1-3 (14% [143/1032] placebo vs. 13% [114/885] and 14% [71/504] for erenumab 70 and 140 mg, respectively) regardless of baseline BP category. The adverse event (AE) profile of erenumab was similar across CV risk categories throughout the long-term analysis. Erenumab-treated patients with high and moderate 10-year CV risk (N = 107) did not experience any ICCAEs during the double-blind treatment period; there was a single ICCAE (a cerebral dural venous sinus thrombosis) observed in the low-risk erenumab group (N = 273). There were no increases in AEs during the long-term extensions of up to 5 years (N = 2499; 3482 patient-years of exposure to erenumab) with exposure-adjusted incidence rates of cardio/cerebrovascular disorder AEs of 0.4, 0.5, 0.0, and 1.1 (per 100 patient-years) for no risk factor (N = 1805), low (N = 492), moderate (N = 121), and high (N = 81) 10-year CV risk groups, respectively.

Cryosectioned tissues from snap-frozen samples offer the advantage of preserving proteins at the cellular and subcellular levels and maintaining overall cell integrity in the tissue of interest without the use of chemical fixatives. To prevent specific or nonspecific degradation of proteins by autolytic and/or proteolytic processes, it is common practice to immediately store frozen tissue sections obtained from a cryostat under cryogenic conditions, for example −80°C. Our laboratory recently challenged this widely held belief by extracting proteins from brain tissue samples that were archived for 1 day, 1 week, 1 month, and 6 months at various storage conditions (frozen, ambient, or desiccated) without the use of chemical fixatives. Our results from immunofluorescent stains, immunoperoxidase stains, silver stains, and Western blot analyses demonstrated that snap-frozen, heat-dried tissue sections stored and desiccated at ambient laboratory conditions are comparable to frozen samples stored up to 6 months. Keywordsprotein snap-frozen heat-dried; desiccated; protein extraction; molecular biology; immunohistochemistryThe study design and analysis of ex vivo tissue samples determines the method by which the specimen is processed and preserved. Tissues used for morphologic or immunohistochemical analyses are frequently fixed with a common chemical fixative, such as formalin, and are embedded in paraffin. While formalin-fixed paraffin-embedded specimens are well preserved and conveniently stored at ambient room temperatures, the cross-linking and sulfide bond formation caused by the fixative make them less compatible with current molecular techniques. 1-3 Hence, epitope retrieval methodologies are used to recover the antigens; but recovery in some instances can be less than optimal. 1,4,5 An alternative to the use of fixatives is the centuryold method of freezing the tissue at subzero temperatures. Freezing the tissue provides a The widely accepted belief that snap-freezing any tissue adequately preserves protein integrity at cellular and subcellular levels has served as the gold standard for molecular analysis. [1][2][3]9 Ever since Altmann 7 described cellular degradation in the form of proteolysis, the conventional method of cryogenic storage of frozen tissues has been used, and few deviations from this practice have been reported. However, the question remains whether sectioned tissue samples need to be maintained at freezing temperatures to preserve protein integrity. Challenging conventional practice, our laboratory asked whether proteins can be extracted from tissues, specifically brain sections, that were previously snap-frozen and stored at various conditions for a month or longer. MATERIALS AND METHODS Tissue AcquisitionBrain tissue was harvested from rats (n = 5) immediately after intravenous euthanasia (1.0 mL Euthasol I.C., Delmarva Laboratories Inc, Greenland, NH). Tissues were snap-frozen at −55°C in methylbutane cooled with dry ice. The tissue samples were submerged in the cryogenic solution fo...