Mohamed M. Ali

Anatomy and embryology of the lateral sellar compartment (cavernous sinus) medial wall

In order to study the microanatomy of the lateral sellar compartment (cavernous sinus) medial wall, serial histological sections of human fetuses and adults, as well as dissections under operative microscope, were performed. The results were compared to high resolution Magnetic Resonance Images in human, to microdissections and to serial histological sections in adult nonhuman primates (Papio Cynocephalus anubis). We were able to show that the sellar compartment and both lateral sellar osteodural compartments are not separated from each other by a dural wall, but by a more or less dense, interrupted, fibrous tissue which derived from the mesenchyme surrounding the hypophysis, carotid artery, cranial nerves and venous channels. In the human fetus, the previous mentioned structures are located in a unique interperiosteodural space. Histoarchitecture of the superior and lateral wall dura-mater was different from the underlying mesenchyme derived connective tissue and was easily distinguished through histological examination. These findings correspond to MRI data. We conclude that there is no medial dural wall limiting the lateral sellar compartment (cavernous sinus), both parasellar and the hypophyseal compartment should be considered as a unique extradural space. The only dense connective tissue surrounding the pituitary gland is its own glandular capsule and the periosteum.

Genistein increases estrogen receptor beta expression in prostate cancer via reducing its promoter methylation.

Genistein has protective effects against prostate cancer (PCa) but whether this protection involves an estrogen receptor (ER) β dependent mechanism has yet to be elucidated. ER-β has a tumor suppressor role in PCa and its levels decline with cancer progression which was linked to ER-β promoter hypermethylation. Genistein has been suggested to have demethylating activities in cancer. However, the ability of genistein to reverse ER-β promoter hypermethylation in PCa has not been studied. In addition, there are great discrepancies among studies that examined the effect of genistein on ER-β gene expression. Therefore, we sought to explore effects of genistein on ER-β promoter methylation as a mechanism of modulating ER-β expression using three PCa cell lines, LNCaP, LAPC-4 and PC-3. We also examined the role of ER-β in mediating the preventive action of genistein. Our data demonstrated that genistein at physiological ranges (0.5-10 μmol/L) reduced ER-β promoter methylation significantly with corresponding dose-dependent increases in ER-β expression in LNCaP and LAPC-4 but not in PC-3 cells, which could be attributed to the low basal levels of ER-β promoter methylation in PC-3 cell line. Genistein induced phosphorylation, nuclear translocation and transcriptional activity of ER-β in all three PCa cell lines. Inhibitory effects of genistein on LAPC-4 and PC-3 cell proliferation were diminished using a specific ER-β antagonist. In conclusion, genistein and ER-β act together to prevent PCa cell proliferation; genistein increases ER-β levels via reducing its promoter methylation and ER-β, in turn, mediates the preventive action of genistein.

A 3-O-Sulfated Heparan Sulfate Binding Peptide Preferentially Targets Herpes Simplex Virus 2-Infected Cells

ABSTRACT Herpes simplex virus 2 (HSV-2) is the primary cause of genital herpes, which is one of the most common sexually transmitted viral infections worldwide and a major cofactor for human immunodeficiency virus infection. The lack of an effective vaccine or treatment and the emergence of drug-resistant strains highlight the need for developing new antivirals for HSV-2. Here, we demonstrate that a low-molecular-weight peptide isolated against 3-O-sulfated heparan sulfate (3-OS HS) can efficiently block HSV-2 infection. Treatment with the peptide inhibited viral entry and cell-to-cell spread both in vitro and in vivo using a mouse model of genital HSV-2 infection. Quite interestingly, the peptide showed a preferential binding to HSV-2-infected cells, with more than 200% increased binding compared to uninfected cells. Our additional results show that heparan sulfate expression is upregulated by 25% upon HSV-2 infection, which is a significant new finding that could be exploited for designing new diagnostic tests and treatment strategies against HSV-2-infected cells. In addition, our results also raise the possibility that 3-OS HS modifications within HS may be upregulated even more to accommodate for a significantly higher increase in the peptide binding to the infected cells.

Reduced vasodilator function following acute resistance exercise in obese women

Obesity contributes to stress induced impairments in endothelium-dependent vasodilation (EDV), a precursor to atherosclerosis. Since obesity is associated with inflammation and oxidative stress, we sought to determine if a single bout of strenuous weight lifting (SWL) reduces EDV among sedentary obese adults. Participants included 9 obese (OB) (BMI 30.0–40.0 kg/m2) and 8 lean (LN) (BMI 18.5–24.9 kg/m2) sedentary young women. All participants underwent a single bout of SWL using a progressive leg-press protocol. Brachial artery flow-mediated dilation (FMD) (an index of EDV) was determined using ultrasonography before and after SWL. Sublingual nitroglycerin (NTG) was used to determine brachial artery endothelium-independent vasodilation following SWL. Brachial artery FMD was significantly reduced in OB and LN women (LN: 6.4 ± 1.6%, p = 0.22) after SWL. There was no difference in the magnitude of change pre- and post-SWL between groups (OB: −2.4 ± 0.6% and LN: −2.2 ± 1.6%, p = 0.84). Dilation to NTG was lower in OB (21.6 ± 1.3%) compared to LN women (27.6 ± 2.1%, p = 0.02) and associated with body weight (r = −0.70, p = 0.01). These data suggest that EDV is reduced in woman after acute resistance exercise. Dilations to NTG were lower in obese compared to lean woman and associated with body weight suggesting that changes in sensitivity of blood vessels to NO occurs during obesity. These findings may be important for understanding vascular risk following acute exercise in obesity.

Massage therapy restores peripheral vascular function after exertion.

OBJECTIVE To determine if lower extremity exercise-induced muscle injury reduces vascular endothelial function of the upper extremity and if massage therapy (MT) improves peripheral vascular function after exertion-induced muscle injury. DESIGN Randomized, blinded trial with evaluations at 90 minutes, 24 hours, 48 hours, and 72 hours. SETTING Clinical research center. PARTICIPANTS Sedentary young adults (N=36) were randomly assigned to 1 of 3 groups: (1) exertion-induced muscle injury and MT (n=15; mean age ± SE, 26.6 ± 0.3); (2) exertion-induced muscle injury only (n=10; mean age ± SE, 23.6 ± 0.4), and (3) MT only (n=11; mean age ± SE, 25.5 ± 0.4). INTERVENTION Participants were assigned to exertion-induced muscle injury only (a single bout of bilateral, eccentric leg press exercise), MT only (30-min lower extremity massage using Swedish technique), or exertion-induced muscle injury and MT. MAIN OUTCOME MEASURES Brachial artery flow-mediated dilation (FMD) was determined by ultrasound at each time point. Nitroglycerin (NTG)-induced dilation was also assessed (0.4 mg). RESULTS Brachial FMD increased from baseline in the exertion-induced muscle injury and MT group and the MT only group (7.38%±.18% to 9.02%±.28%, P<.05 and 7.77%±.25% to 10.2%±.22%, P<.05, respectively) at 90 minutes and remained elevated until 72 hours. In the exertion-induced muscle injury only group, FMD was reduced from baseline at 24 and 48 hours (7.78%±.14% to 6.75%±.11%, P<.05 and 6.53%±.11%, P<.05, respectively) and returned to baseline after 72 hours. Dilations of NTG were similar over time. CONCLUSIONS Our results suggest that MT attenuates impairment of upper extremity endothelial function resulting from lower extremity exertion-induced muscle injury in sedentary young adults.

An Important Role for Syndecan-1 in Herpes Simplex Virus Type-1 Induced Cell-to-Cell Fusion and Virus Spread

Herpes simplex virus type-1 (HSV-1) is a common human pathogen that relies heavily on cell-to-cell spread for establishing a lifelong latent infection. Molecular aspects of HSV-1 entry into host cells have been well studied; however, the molecular details of the spread of the virus from cell-to-cell remain poorly understood. In the past, the role of heparan sulfate proteoglycans (HSPG) during HSV-1 infection has focused solely on the role of HS chains as an attachment receptor for the virus, while the core protein has been assumed to perform a passive role of only carrying the HS chains. Likewise, very little is known about the involvement of any specific HSPGs in HSV-1 lifecycle. Here we demonstrate that a HSPG, syndecan-1, plays an important role in HSV-1 induced membrane fusion and cell-to-cell spread. Interestingly, the functions of syndecan-1 in fusion and spread are independent of the presence of HS on the core protein. Using a mutant CHO-K1 cell line that lacks all glycosaminoglycans (GAGs) on its surface (CHO-745) we demonstrate that the core protein of syndecan-1 possesses the ability to modulate membrane fusion and viral spread. Altogether, we identify a new role for syndecan-1 in HSV-1 pathogenesis and demonstrate HS-independent functions of its core protein in viral spread.

Short-term regular aerobic exercise reduces oxidative stress produced by acute high intraluminal pressure in the adipose microvasculature

High blood pressure has been shown to elicit impaired dilation in the vasculature. The purpose of this investigation was to elucidate the mechanisms through which high pressure may elicit vascular dysfunction and determine the mechanisms through which regular aerobic exercise protects arteries against high pressure. Male C57BL/6J mice were subjected to 2 wk of voluntary running (~6 km/day) for comparison with sedentary controls. Hindlimb adipose resistance arteries were dissected from mice for measurements of flow-induced dilation (FID; with or without high intraluminal pressure exposure) or protein expression of NADPH oxidase II (NOX II) and superoxide dismutase (SOD). Microvascular endothelial cells were subjected to high physiological laminar shear stress (20 dyn/cm2) or static condition and treated with ANG II + pharmacological inhibitors. Cells were analyzed for the detection of ROS or collected for Western blot determination of NOX II and SOD. Resistance arteries from exercised mice demonstrated preserved FID after high pressure exposure, whereas FID was impaired in control mouse arteries. Inhibition of ANG II or NOX II restored impaired FID in control mouse arteries. High pressure increased superoxide levels in control mouse arteries but not in exercise mouse arteries, which exhibited greater ability to convert superoxide to H2O2 Arteries from exercised mice exhibited less NOX II protein expression, more SOD isoform expression, and less sensitivity to ANG II. Endothelial cells subjected to laminar shear stress exhibited less NOX II subunit expression. In conclusion, aerobic exercise prevents high pressure-induced vascular dysfunction through an improved redox environment in the adipose microvasculature.NEW & NOTEWORTHY We describe potential mechanisms contributing to aerobic exercise-conferred protection against high intravascular pressure. Subcutaneous adipose microvessels from exercise mice express less NADPH oxidase (NOX) II and more superoxide dismutase (SOD) and demonstrate less sensitivity to ANG II. In microvascular endothelial cells, shear stress reduced NOX II but did not influence SOD expression.

Nox2 contributes to hyperinsulinemia-induced redox imbalance and impaired vascular function

Insulin resistance promotes vascular endothelial dysfunction and subsequent development of cardiovascular disease. Previously we found that skeletal muscle arteriolar flow-induced dilation (FID) was reduced following a hyperinsulinemic clamp in healthy adults. Therefore, we hypothesized that hyperinsulinemia, a hallmark of insulin resistance, contributes to microvascular endothelial cell dysfunction via inducing oxidative stress that is mediated by NADPH oxidase (Nox) system. We examined the effect of insulin, at levels that are comparable with human hyperinsulinemia on 1) FID of isolated arterioles from human skeletal muscle tissue in the presence and absence of Nox inhibitors and 2) human adipose microvascular endothelial cell (HAMECs) expression of nitric oxide (NO), endothelial NO synthase (eNOS), and Nox-mediated oxidative stress. In six lean healthy participants (mean age 25.5±1.6 y, BMI 21.8±0.9), reactive oxygen species (ROS) were increased while NO and arteriolar FID were reduced following 60 min of ex vivo insulin incubation. These changes were reversed after co-incubation with the Nox isoform 2 (Nox2) inhibitor, VAS2870. In HAMECs, insulin-induced time-dependent increases in Nox2 expression and P47phox phosphorylation were echoed by elevations of superoxide production. In contrast, phosphorylation of eNOS and expression of superoxide dismutase (SOD2 and SOD3) isoforms showed a biphasic response with an increased expression at earlier time points followed by a steep reduction phase. Insulin induced eNOS uncoupling that was synchronized with a drop of NO and a surge of ROS production. These effects were reversed by Tempol (SOD mimetic), Tetrahydrobiopterin (BH4; eNOS cofactor), and VAS2870. Finally, insulin induced nitrotyrosine formation which was reversed by inhibiting NO or superoxide generation. In conclusions, hyperinsulinemia may reduce FID via inducing Nox2-mediated superoxide production in microvascular endothelial cells which reduce the availability of NO and enhances peroxynitrite formation. Therefore, the Nox2 pathway should be considered as a target for the prevention of oxidative stress-associated endothelial dysfunction during hyperinsulinemia.

Circuit Resistance Training Attenuates Acute Exertion-Induced Reductions in Arterial Function but Not Inflammation in Obese Women

BACKGROUND Cardiovascular disease (CVD) is a leading cause of preventable death among young women in the United States. Habitual resistance exercise training is known to have beneficial effects on endothelial function and CVD risk factors, including obesity; however, previous studies show that acute resistance exercise impairs endothelial function in obese adults who are sedentary, a response that may be linked to inflammation. We sought to determine if circuit-based resistance training (CRT) attenuates acute resistance exercise-induced reductions in endothelial function in a population of young, obese, sedentary women and whether or not inflammation plays a role in this response. METHODS Eighteen obese [body mass index (BMI) 30.0-40.0 kg · m(-2)] young premenopausal women were randomly assigned to either a CRT group or a no-exercise control group (CON). Conduit artery endothelial function was assessed using brachial artery flow-mediated dilation (FMD) determined by ultrasound before and after a single bout of strenuous weightlifting (SWL). In addition, circulating inflammatory mediators (tumor necrosis factor-α and C-reactive protein), blood pressure, fasting blood lipids, glucose, waist circumference, body composition, and aerobic capacity were assessed. RESULTS Among participants randomized to the CRT group, 8 weeks of training led to considerable increases in FMD after SWL (P=0.001) compared to the CON group. However, no significant differences between the groups were observed in circulating inflammatory mediators, blood pressure, fasting blood lipids, or other physical and physiological characteristics. CONCLUSIONS This study shows that CRT alleviates acute exertion-induced reductions in endothelial function among obese sedentary women in the absence of changes in inflammation.

Short-term high salt intake reduces brachial artery and microvascular function in the absence of changes in blood pressure.

Objectives: The aims of this study were to test the hypothesis that short-term high salt intake reduces macrovascular and microvascular endothelial function in the absence of changes in blood pressure and to determine whether acute exercise restores endothelial function after high salt in women. Materials and methods: Twelve women were administered high salt (11 g of sodium chloride for 7 days) and then underwent a weightlifting session. Brachial artery flow-mediated dilation and nitroglycerin dilation were measured with ultrasound at baseline, after high salt, and after weightlifting. Subcutaneous fat tissue biopsies were obtained at baseline, after high salt, and after weightlifting. Resistance arteries from biopsies were cannulated for vascular reactivity measurements in response to flow [flow-induced dilation (FID)] and acetylcholine. Results: Blood pressure was similar before and after high salt diet. Brachial flow-mediated dilation was reduced after high salt diet but was not affected by acute weightlifting. Brachial nitroglycerin dilations were similar before and after high salt. FID and acetylcholine-induced dilation of resistance arteries were similar to that of before and after high salt diet. FID and acetylcholine-induced dilation was not altered by weightlifting after high salt diet. However, N&ohgr;-nitro-L-arginine methyl ester significantly reduced FID at baseline and after exercise but had no effect dilator reactivity after high salt diet alone. Conclusion: These data suggest that high salt intake reduces brachial artery endothelial function and switches the mediator of vasodilation in the microcirculation to a non-nitric oxide-dependent mechanism in healthy adults and acute exercise may switch the dilator mechanism back to nitric oxide during high salt diet.