Sex-Specific Acute Cerebrovascular Responses to Photothrombotic Stroke in Mice.

Mechanisms underlying cerebrovascular stroke outcomes are poorly understood, and the effects of biological sex on cerebrovascular regulation post-stroke have yet to be fully comprehended. Here, we explore the overlapping roles of gonadal sex hormones and rho-kinase (ROCK), two important modulators of cerebrovascular tone, on the acute cerebrovascular response to photothrombotic (PT) focal ischemia in mice. Male mice were gonadectomized and female mice were ovariectomized to remove gonadal hormones, whereas control ("intact") animals received a sham surgery prior to stroke induction. Intact wild-type (WT) males showed a delayed drop in cerebral blood flow (CBF) compared with intact WT females, whereby maximal CBF drop was observed 48 h following stroke. Gonadectomy in males did not alter this response. However, ovariectomy in WT females produced a "male-like" phenotype. Intact Rock2+/- males also showed the same phenotypic response, which was not altered by gonadectomy. Alternatively, intact Rock2+/- females showed a significant difference in CBF values compared with intact WT females, displaying higher CBF values immediately post-stroke and showing a maximal CBF drop 48 h post-stroke. This pattern was not altered by ovariectomy. Altogether, these data illustrate sex differences in acute CBF responses to PT stroke, which seem to involve gonadal female sex hormones and ROCK2. Overall, this study provides a framework for exploring sex differences in acute CBF responses to focal ischemic stroke in mice.

Pharmacokinetic modeling of dynamic contrast-enhanced (DCE)-MRI in PI-RADS category 3 peripheral zone lesions: preliminary study evaluating DCE-MRI as an imaging biomarker for detection of clinically significant prostate cancers.

PURPOSE: To determine if pharmacokinetic modeling of DCE-MRI can diagnose CS-PCa in PI-RADS category 3 PZ lesions with subjective negative DCE-MRI. MATERIALS AND METHODS: In the present IRB approved, bi-institutional, retrospective, case-control study, we identified 73 men with 73 PZ PI-RADS version 2.1 category 3 lesions with MRI-directed-TRUS-guided targeted biopsy yielding: 12 PZ CS-PCa (ISUP Grade Group 2; N = 9, ISUP 3; N = 3), 27 ISUP 1 PCa and 34 benign lesions. An expert blinded radiologist segmented lesions on ADC and DCE images; segmentations were overlayed onto pharmacokinetic DCE-MRI maps. Mean values were compared between groups using univariate analysis. Diagnostic accuracy was assessed by ROC. RESULTS: There were no differences in age, PSA, PSAD or clinical stage between groups (p = 0.265-0.645). Mean and 10th percentile ADC did not differ comparing CS-PCa to ISUP 1 PCa and benign lesions (p = 0.376 and 0.598) but was lower comparing ISUP ≥ 1 PCa to benign lesions (p < 0.001). Mean Ktrans (p = 0.003), Ve (p = 0.003) but not Kep (p = 0.387) were higher in CS-PCa compared to ISUP 1 PCa and benign lesions. There were no differences in DCE-MRI metrics comparing ISUP ≥ 1 PCa and benign lesions (p > 0.05). AUC for diagnosis of CS-PCa using Ktrans and Ve were: 0.69 (95% CI 0.52-0.87) and 0.69 (0.49-0.88). CONCLUSION: Pharmacokinetic modeling of DCE-MRI parameters in PI-RADS category 3 lesions with subjectively negative DCE-MRI show significant differences comparing CS-PCa to ISUP 1 PCa and benign lesions, in this study outperforming ADC. Studies are required to further evaluate these parameters to determine which patients should undergo targeted biopsy for PI-RADS 3 lesions.

Injectable human recombinant collagen matrices limit adverse remodeling and improve cardiac function after myocardial infarction.

Despite the success of current therapies for acute myocardial infarction (MI), many patients still develop adverse cardiac remodeling and heart failure. With the growing prevalence of heart failure, a new therapy is needed that can prevent remodeling and support tissue repair. Herein, we report on injectable recombinant human collagen type I (rHCI) and type III (rHCIII) matrices for treating MI. Injecting rHCI or rHCIII matrices in mice during the late proliferative phase post-MI restores the myocardium's mechanical properties and reduces scar size, but only the rHCI matrix maintains remote wall thickness and prevents heart enlargement. rHCI treatment increases cardiomyocyte and capillary numbers in the border zone and the presence of pro-wound healing macrophages in the ischemic area, while reducing the overall recruitment of bone marrow monocytes. Our findings show functional recovery post-MI using rHCI by promoting a healing environment, cardiomyocyte survival, and less pathological remodeling of the myocardium.

Hyperfiltration in ubiquitin C-terminal hydrolase L1-deleted mice.

Neuronal ubiquitin C-terminal hydrolase L1 (UCHL1) is a deubiquitinating enzyme that maintains intracellular ubiquitin pools and promotes axonal transport. Uchl1 deletion in mice leads to progressive axonal degeneration, affecting the dorsal root ganglion that harbors axons emanating to the kidney. Innervation is a crucial regulator of renal hemodynamics, though the contribution of neuronal UCHL1 to this is unclear. Immunofluorescence revealed significant neuronal UCHL1 expression in mouse kidney, including periglomerular axons. Glomerular filtration rate trended higher in 6-week-old Uchl1-/- mice, and by 12 weeks of age, these displayed significant glomerular hyperfiltration, coincident with the onset of neurodegeneration. Angiotensin converting enzyme inhibition had no effect on glomerular filtration rate of Uchl1-/- mice indicating that the renin-angiotensin system does not contribute to the observed hyperfiltration. DCE-MRI revealed increased cortical renal blood flow in Uchl1-/- mice, suggesting that hyperfiltration results from afferent arteriole dilation. Nonetheless, hyperglycemia, cyclooxygenase-2, and nitric oxide synthases were ruled out as sources of hyperfiltration in Uchl1-/- mice as glomerular filtration rate remained unchanged following insulin treatment, and cyclooxygenase-2 and nitric oxide synthase inhibition. Finally, renal nerve dysfunction in Uchl1-/- mice is suggested given increased renal nerve arborization, decreased urinary norepinephrine, and impaired vascular reactivity. Uchl1-deleted mice demonstrate glomerular hyperfiltration associated with renal neuronal dysfunction, suggesting that neuronal UCHL1 plays a crucial role in regulating renal hemodynamics.

Vascular Smooth Muscle-Specific EP4 Receptor Deletion in Mice Exacerbates Angiotensin II-Induced Renal Injury.

AIMS: Cyclooxygenase inhibition by non-steroidal anti-inflammatory drugs is contraindicated in hypertension, as it may reduce glomerular filtration rate (GFR) and renal blood flow. However, the identity of the specific eicosanoid and receptor underlying these effects is not known. We hypothesized that vascular smooth muscle prostaglandin E2 (PGE2) E-prostanoid 4 (EP4) receptor deletion predisposes to renal injury via unchecked vasoconstrictive actions of angiotensin II (AngII) in a hypertension model. Mice with inducible vascular smooth muscle cell (VSMC)-specific EP4 receptor deletion were generated and subjected to AngII-induced hypertension. RESULTS: EP4 deletion was verified by PCR of aorta and renal vessels, as well as functionally by loss of PGE2-mediated mesenteric artery relaxation. Both AngII-treated groups became similarly hypertensive, whereas albuminuria, foot process effacement, and renal hypertrophy were exacerbated in AngII-treated EP4VSMC-/- but not in EP4VSMC+/+ mice and were associated with glomerular scarring, tubulointerstitial injury, and reduced GFR. AngII-treated EP4VSMC-/- mice exhibited capillary damage and reduced renal perfusion as measured by fluorescent bead microangiography and magnetic resonance imaging, respectively. NADPH oxidase 2 (Nox2) expression was significantly elevated in AngII-treated EP4-/- mice. EP4-receptor silencing in primary VSMCs abolished PGE2 inhibition of AngII-induced Nox2 mRNA and superoxide production. INNOVATION: These data suggest that vascular EP4 receptors buffer the actions of AngII on renal hemodynamics and oxidative injury. CONCLUSION: EP4 agonists may, therefore, protect against hypertension-associated kidney damage. Antioxid. Redox Signal. 25, 642-656.

MRI evaluation of small (<4cm) solid renal masses: multivariate modeling improves diagnostic accuracy for angiomyolipoma without visible fat compared to univariate analysis.

OBJECTIVE: To assess MRI for diagnosis of angiomyolipoma without visible fat (AMLwvf). MATERIAL AND METHODS: With IRB approval, a retrospective study in consecutive patients with contrast-enhanced (CE)-MRI and <4 cm solid renal masses from 2002-2013 was performed. Ten AMLwvf were compared to 77 RCC; 33 clear cell (cc), 35 papillary (p), 9 chromophobe (ch). A blinded radiologist measured T2W signal-intensity ratio (SIR), chemical-shift (CS) SI-index and area under CE-MRI curve (CE-AUC). Regression modeling and ROC analysis was performed. RESULTS: T2W-SIR was lower in AMLwvf (0.64 ± 0.12) compared to cc-RCC (1.37 ± 0.30, p < 0.001), ch-RCC (0.94 ± 0.19, p = 0.005) but not p-RCC (0.74 ± 0.17, p = 0.2). CS-SI index was higher in AMLwvf (16.1 ± 31.5 %) compared to p-RCC (-5.2 ± 26.1 %, p = 0.02) but not ch-RCC (3.0 ± 12.5 %, p = 0.1) or cc-RCC (7.7 ± 17.9 %,p = 0.1). CE-AUC was higher in AMLwvf (515.7 ± 144.7) compared to p-RCC (154.5 ± 92.8, p < 0.001) but not ch-RCC (341.5 ± 202.7, p = 0.07) or cc-RCC (520.9 ± 276.9, p = 0.95). Univariate ROC-AUC were: T2SIR = 0.86 (CI 0.77-0.96); CE-AUC = 0.76 (CI 0.65-0.87); CS-SI index = 0.66 (CI 0.4.3-0.85). Logistic regression models improved ROC-AUC, A) T2 SIR + CE-AUC = 0.97 (CI 0.93-1.0) and T2 SIR + CS-SI index = 0.92 (CI 0.84-0.99) compared to univariate analyses (p < 0.05). The optimal sensitivity/specificity of T2SIR + CE-AUC and T2SIR + CS-SI index were 100/88.8 % and 60/97.4 %. CONCLUSION: MRI, using multi-variate modelling, is accurate for diagnosis of AMLwvf. KEY POINTS: • AMLwvf are difficult to prospectively diagnose with imaging. • MRI findings associated with AMLwvf overlap with various RCC subtypes. • T2W-SI combined with chemical-shift SI-index is specific for AMLwvf but lacks sensitivity. • T2W-SI combined with AUC CE-MRI is sensitive and specific for AMLwvf. • Models incorporating two or more findings are more accurate than univariate analysis.

Glucocorticoids modulate tumor radiation response through a decrease in tumor oxygen consumption.

PURPOSE: We hypothesized that glucocorticoids may enhance tumor radiosensitivity by increasing tumor oxygenation (pO(2)) through inhibition of mitochondrial respiration. EXPERIMENTAL DESIGN: The effect of three glucocorticoids (hydrocortisone, dexamethasone, and prednisolone) on pO(2) was studied in murine TLT liver tumors and FSaII fibrosarcomas. At the time of maximum pO(2) (t(max), 30 min after administration), perfusion, oxygen consumption, and radiation sensitivity were studied. Local pO(2) measurements were done using electron paramagnetic resonance. The oxygen consumption rate of tumor cells after in vivo glucocorticoid administration was measured using high-frequency electron paramagnetic resonance. Tumor perfusion and permeability measurements were assessed by dynamic contrast-enhanced magnetic resonance imaging. RESULTS: All glucocorticoids tested caused a rapid increase in pO(2). At t(max), tumor perfusion decreased, indicating that the increase in pO(2) was not caused by an increase in oxygen supply. Also at t(max), global oxygen consumption decreased. When irradiation (25 Gy) was applied at t(max), the tumor radiosensitivity was enhanced (regrowth delay increased by a factor of 1.7). CONCLUSION: These results show the potential usefulness of the administration of glucocorticoids before irradiation.

Determination of the maturity and functionality of tumor vasculature by MRI: correlation between BOLD-MRI and DCE-MRI using P792 in experimental fibrosarcoma tumors.

Using hypercapnia and carbogen as functional markers of vessel maturation and function, we compared blood oxygen level-dependent (BOLD) contrast with standard dynamic contrast-enhanced (DCE)-MRI quantitative parameters in murine fibrosarcoma. Our results show that there was no correlation between vessel maturity and contrast-agent uptake rate (K(in) (Trans)) or contrast agent efflux rate (k(ep)). In addition, DCE-MRI provided higher estimates of the fraction of functional tumor compared to BOLD-MRI. The two putative markers of regional vascular density, i.e., the magnitude of BOLD signal change during carbogen challenge (VF) and the fractional plasma volume found by DCE-MRI (V(p)), were only weakly correlated (r(2) = 0.02-0.14). Furthermore, VF showed no correlation with K(in) (Trans). A positive correlation was observed (r(2) = 0.75) between mean tumor VF and k(ep), but only when averaged over the whole tumor (which includes tumor regions completely unperfused by the gadolinium (Gd) contrast agent). This would merely reveal a relationship between perfusion status and the capacity to respond to carbogen breathing. In conclusion, characterizations of tumor microvasculature imaging using BOLD-MRI and DCE-MRI appear to be largely complementary, given the weak correlations between their corresponding derived parameters.

Measurement of xenon diffusing capacity in the rat lung by hyperpolarized 129Xe MRI and dynamic spectroscopy in a single breath-hold.

We used the dual capability of hyperpolarized 129Xe for spectroscopy and imaging to develop new measures of xenon diffusing capacity in the rat lung that (analogously to the diffusing capacity of carbon monoxide or DLCO) are calculated as a product of total lung volume and gas transfer rate constants divided by the pressure gradient. Under conditions of known constant pressure breath-hold, the volume is measured by hyperpolarized 129Xe MRI, and the transfer rate is measured by dynamic spectroscopy. The new quantities (xenon diffusing capacity in lung parenchyma (DLXeLP)), xenon diffusing capacity in RBCs (DLXeRBC), and total lung xenon diffusing capacity (DLXe)) were measured in six normal rats and six rats with lung inflammation induced by instillation of fungal spores of Stachybotrys chartarum. DLXeLP, DLXeRBC, and DLXe were 56 +/- 10 ml/min/mmHg, 64 +/- 35 ml/min/mmHg, and 29 +/- 9 ml/min/mmHg, respectively, for normal rats, and 27 +/- 9 ml/min/mmHg, 42 +/- 27 ml/min/mmHg, and 16 +/- 7 ml/min/mmHg, respectively, for diseased rats. Lung volumes and gas transfer times for LP (TtrLP) were 16 +/- 2 ml and 22 +/- 3 ms, respectively, for normal rats and 12 +/- 2 ml and 35 +/- 8 ms, respectively, for diseased rats. Xenon diffusing capacities may be useful for measuring changes in gas exchange associated with inflammation and other lung diseases.

Tumor radiosensitization by antiinflammatory drugs: evidence for a new mechanism involving the oxygen effect.

We hypothesized that nonsteroidal antiinflammatory drugs (NSAIDs) might enhance tumor radiosensitivity by increasing tumor oxygenation (pO2), via either a decrease in the recruitment of macrophages or from inhibition of mitochondrial respiration. The effect of four NSAIDs (diclofenac, indomethacin, piroxicam, and NS-398) on pO2 was studied in murine TLT liver tumors and FSaII fibrosarcomas. At the time of maximum pO2 (t(max), 30 minutes after administration), perfusion, oxygen consumption, and radiation sensitivity were studied. Local pO2 measurements were done using electron paramagnetic resonance. Tumor perfusion and permeability measurements were assessed by dynamic contrast-enhanced magnetic resonance imaging. The oxygen consumption rate of tumor cells after in vivo NSAID administration was measured using high-frequency electron paramagnetic resonance. Tumor-infiltrating macrophage localization was done with immunohistochemistry using CD11b antibody. All the NSAIDs tested caused a rapid increase in pO2. At t(max), tumor perfusion decreased, indicating that the increase in pO2 was not caused by an increase in oxygen supply. Also at t(max), global oxygen consumption decreased but the amount of tumor-infiltrating macrophages remained unchanged. Our study strongly indicates that the oxygen effect caused by NSAIDs is primarily mediated by an effect on mitochondrial respiration. When irradiation (18 Gy) was applied at t(max), the tumor radiosensitivity was enhanced (regrowth delay increased by a factor of 1.7). These results show the potential utility of an acute administration of NSAIDs for radiosensitizing tumors, and shed new light on the mechanisms of NSAID radiosensitization. These results also provide a new rationale for the treatment schedule when combining NSAIDs and radiotherapy.