In vitro measurements of hemodynamic forces and their effects on endothelial cell mechanics at the sub-cellular level.

This paper presents micro-particle tracking velocimetry measurements over cultured bovine aortic endothelial cell monolayers in microchannels. The objective was to quantify fluid forces and cell morphology at the sub-cellular scale for monolayers subjected to steady shear rates of 5, 10, and 20 dyn/cm2. The ultimate goal of this study was to develop an experimental methodology for in vitro detailed study of physiologically realistic healthy and diseased conditions. Cell topography, shear stress, and pressure distributions were calculated from sets of velocity fields made in planes parallel to the microchannel wall. For each experiment, measurements were made in 3 h intervals for 18 h. It was found that there is a three-dimensional change in cell morphology as a result of applied shear stress. That is, cells flatten and become more wedge shaped in the stream direction while conserving volume by spreading laterally, i.e., in the cross-stream direction. These changes in cell morphology are directly related to local variations in fluid loading, i.e., shear stress and pressure. This paper describes the first flow measurements over a confluent layer of endothelial cells that are spatially resolved at the sub-cellular scale with a simultaneous temporal resolution to quantify the response of cells to fluid loading.

Dexamethasone promotes long-term functional recovery of neuromuscular junction in a murine model of tourniquet-induced ischaemia-reperfusion.

AIM: Tourniquet-induced ischaemia and subsequent reperfusion cause serious ischaemia-reperfusion (IR) injury in the neuromuscular junction (NMJ) and skeletal muscle. Here, we investigated whether dexamethasone (Dex) promotes long-term functional recovery of the NMJ and skeletal muscle in tourniquet-induced hindlimb IR. METHODS: Unilateral hindlimb of C57/BL6 mice was subjected to 3 h of ischaemia following 6 weeks of reperfusion (6-wk IR). Dex treatment began on the day of IR induction and lasted for different periods. Sciatic nerve-stimulated gastrocnemius muscle contraction was detected in situ. Function of the NMJ was measured in situ using electrophysiological recording of the miniature endplate potential (mEPP) and endplate potential (EPP). Western blot was used to detect protein expression of nicotinic acetylcholine receptors (nAChRs) in gastrocnemius muscles. RESULTS: Gastrocnemius muscle contraction in mice with 6-wk IR was about 60% of normal skeletal muscle contraction recorded in age-matched sham mice. The amplitude of the mEPP and EPP was lower in mice with 6-wk IR, compared to sham mice. Dex treatment for 1 or 3 days did not restore the function of the NMJ and improve gastrocnemius muscle contraction in mice with 6-wk IR. Dex treatment for 1 week exerted a maximum effect on improving the function of the NMJ and skeletal muscle, with the effect of Dex gradually lessening with prolonged Dex treatment. There are no significant differences in protein expression of nAChR-α1 and nAChR-ß1 subunits in the gastrocnemius muscle among all groups. CONCLUSION: Dex promotes repair of the NMJ and subsequently restores skeletal muscle contractile function in tourniquet-induced 6-wk IR.

Spontaneous and coherent anti-Stokes Raman spectroscopy of human gastrocnemius muscle biopsies in CH-stretching region for discrimination of peripheral artery disease.

Peripheral Artery Disease (PAD) is a common manifestation of atherosclerosis, characterized by lower leg ischemia and myopathy in association with leg dysfunction. In the present study, Spontaneous and coherent anti-Stokes Raman scattering (CARS) spectroscopic techniques in CH-stretching spectral region were evaluated for discriminating healthy and diseased tissues of human gastrocnemius biopsies of control and PAD patients. Since Raman signatures of the tissues in the fingerprint region are highly complex and CH containing moieties are dense, CH-stretching limited spectral range was used to classify the diseased tissues. A total of 181 Raman spectra from 9 patients and 122 CARS spectra from 12 patients were acquired. Due to the high dimensionality of the data in Raman and CARS measurements, principal component analysis (PCA) was first performed to reduce the dimensionality of the data (6 and 9 principal scores for Raman and CARS, respectively) in the CH-stretching region, followed by a discriminant function analysis (DFA) to classify the samples into different categories based on disease severity. The CH2 and CH3 vibrational signatures were observed in the Raman and CARS spectroscopy. Raman and CARS data in conjunction with PCA-DFA analysis were capable of differentiating healthy and PAD gastrocnemius with an accuracy of 85.6% and 78.7%, respectively.

Protein concentration and mitochondrial content in the gastrocnemius predicts mortality rates in patients with peripheral arterial disease.

OBJECTIVE: This study evaluated the hypothesis that protein concentration and mitochondrial content in gastrocnemius biopsies from patients with peripheral arterial disease (PAD) predict mortality rates. BACKGROUND: PAD patients experience advancing myopathy characterized by mitochondrial dysfunction, myofiber degradation, and fibrosis in their ischemic legs, along with increased mortality rates. METHODS: Samples from the gastrocnemius of PAD patients were used for all analyses. Protein concentration was normalized to muscle wet weight, and citrate synthase activity (standard measure of mitochondrial content in cells) was normalized to muscle wet weight and protein concentration. Protein and citrate synthase data were grouped into tertiles and 5-year, all-cause mortality for each tertile was determined with Kaplan-Meier curves and compared by the modified Peto-Peto test. A Cox-regression model for each variable controlled for the effects of clinical characteristics. RESULTS: Of the 187 study participants, 46 died during a mean follow-up of 23.0 months. Five-year mortality rate was highest for patients in the lowest tertile of protein concentration. Mortality was lowest for patients in the middle tertile of citrate synthase activity when normalized to either muscle wet weight or protein concentration. The mortality hazard ratios (HRs) from the Cox analysis were statistically significant for protein concentration normalized to muscle wet weight (lowest vs middle tertile; HR = 2.93; P = 0.008) and citrate synthase normalized to protein concentration (lowest vs middle tertile; HR = 4.68; P = 0.003; and lowest vs highest tertile; HR = 2.36; P = 0.027). CONCLUSIONS: Survival analysis of a contemporaneous population of PAD patients identifies protein and mitochondrial content of their gastrocnemius as predictors of mortality rate.

Endovascular repair of abdominal aortic aneurysm does not improve early survival versus open repair in patients younger than 60 years.

OBJECTIVES: Multiple randomised trials have demonstrated lower perioperative mortality after endovascular aneurysm repair (EVAR) compared to open surgical repair for infrarenal abdominal aortic aneurysms (AAAs). However, in these trials the mortality advantage for EVAR is being lost within 2 years of repair and the patients evaluated are relatively older with no study specifically comparing EVAR and open repair for patients younger than 60 years of age. DESIGN: A retrospective analysis of prospectively collected data. MATERIALS AND METHODS: Patients younger than 60 years of age who underwent EVAR and open surgical repair for elective infrarenal AAA were identified from the 2007-09 National Surgical Quality Improvement Program (NSQIP) - a prospective database maintained at 237 centres across the United States. Univariate and multivariate analyses were performed. RESULTS: Of the 651 patients, 369 (56.7%) underwent EVAR and 282 (43.3%) underwent open repair. Thirty-day mortality for EVAR and open repair were 1.1% and 0.4%, respectively. This was not significantly different on univariate (P = 0.22) as well as multivariate (P = 0.69) analysis after controlling for other co-morbidities. On multivariate analysis, body mass index, history of stroke and bleeding disorder prior to surgery were associated with a higher 30-day mortality after AAA repair (combined open and EVAR). CONCLUSIONS: These contemporary results demonstrate that the 30-day mortality rate after open repair is similar to that after EVAR in patients younger than 60 years with infrarenal AAA.

Factors affecting outcome in proximal abdominal aortic aneurysm repair.

Sixty-five consecutive patients undergoing nonemergent repair of an abdominal aortic aneurysm (AAA) originating above the visceral and/or renal arteries were studied to determine operative results and identify factors influencing outcome of proximal AAA repair. Factors associated with postoperative morbidity were analyzed using multivariate analysis. There were no postoperative deaths, paraplegia/paraparesis, or symptomatic visceral ischemia. Proximal AAA repair can be accomplished with acceptable mortality. If renal artery bypass or reimplantation is anticipated, cold renal perfusion may protect against renal dysfunction. Postoperative pulmonary dysfunction can be reduced by avoiding radial division of the diaphragm.

Analysis of coagulation changes associated with supraceliac aortic crossclamping using thromboelastography.

INTRODUCTION: The etiology of the coagulation changes seen with supraceliac (SC) aortic crossclamping (AXC) remains controversial; both primary fibrinolysis and clotting factor consumption have been implicated. The cause of these changes was investigated with thromboelastography (TEG), a test that measures the viscoelastic properties of thrombus to dynamically assess coagulation and fibrinolysis. METHODS: Eight pigs underwent SC AXC for 30 min; 5 pigs undergoing 30 min of infrarenal (IR) aortic clamping served as controls. Blood was drawn before AXC, before unclamping, and 5 and 60 min after unclamping. Thromboelastography and standard coagulation tests [prothrombin time (PT), partial thromboplastin time (PTT), fibrinogen, and platelet count] were performed. Measured TEG parameters included fibrinolytic index (a measure of fibrinolysis), r value (a reflection of intrinsic coagulation cascade activity), and the alpha angle and K values (measures of the speed of solid clot formation). Repeated measures ANOVA and t test were used for statistical analysis. RESULTS: There was no difference in the fibrinolytic index at any time point between the two groups. Increased activity of the intrinsic coagulation cascade during SC clamping was reflected by a lower R value just before unclamping (12.6 +/- 3.0 vs 20.0 +/- 3.0, P = 0.048) compared to IR AXC. Decreased speed of solid clot formation was noted 5 min after unclamping in the SC group but not the IR group [as defined by an increased K value (ANOVA, P = 0.010) and a decreased alpha angle value (ANOVA, P = 0.005)]. Fibrinogen levels were lower in the SC than in the IR group 5 (P = 0.013) and 60 min after unclamping (P = 0.02), but PT, PTT, and platelets did not differ between the groups at any time points. CONCLUSIONS: Thirty minutes of SC AXC does not result in fibrinolysis. There is increased clotting activity during SC clamping followed by decreased speed of clot formation and decreased fibrinogen levels after unclamping. These changes are consistent with clotting factor consumption.

Retrograde jejunogastric intussusception.

Retrograde gastrointestinal intussusception is a rare entity, most commonly reported after gastric resection and gastrojejunostomy. Its occurrence in the absence of previous gastric resection is extremely unusual, with only four cases reported. All cases were associated with previously placed gastrostomy tubes and implicated these as the inciting factor. We present a fifth case and review the literature. The mechanism of this phenomenon is described and recommendations to prevent this potentially fatal complication are made.

Lymphoepithelial cyst in the pancreas: a case report and review of the literature.

BACKGROUND: Lymphoepithelial cysts of the pancreas constitute a rare clinicopathologic entity. CASE REPORT: We report a case of lymphoepithelial cyst of the pancreas and review the world literature. RESULTS: Lymphoepithelial cysts are true pancreatic cysts lined by squamous epithelium and surrounded by mature lymphoid tissue. The cyst arises typically in middle aged men, and is usually asymptomatic or causes nonspecific abdominal complaints. There is no specific serologic marker for this entity. None of its radiologic characteristics can help differentiate it from other cystic lesions of the pancreas. Fine-needle aspiration cytology may be able to suggest its benign nature and identify it as a true cyst of the pancreas. The outcome after surgical excision is uniformly good with good symptom control and no recurrences. RECOMMENDATIONS: In the symptomatic patient or the asymptomatic patient with acceptable surgical risk a simple cyst excision should be performed after verification of the diagnosis with frozen section. In the asymptomatic patient with a high surgical risk, in whom fine-needle aspiration suggests the diagnosis of a lymphoepithelial cyst, observation of the lesion is recommended. When simple cyst excision is technically not possible, extensive resections/reconstructions should be avoided and drainage/bypass procedures may be considered.

Phosphorus 31 nuclear magnetic resonance spectroscopy suggests a mitochondrial defect in claudicating skeletal muscle.

OBJECTIVE: Decreased oxygen supply is generally accepted as the primary cause of muscle dysfunction in patients with peripheral arterial occlusive disease (PAOD) and intermittent claudication, although reported morphologic changes in the mitochondria of claudicating muscle suggest that impaired energy utilization may also play a role. With the measurement of the phosphate-rich compounds of muscle energy metabolism (adenosinetriphosphate [ATP], adenosinediphosphate [ADP], and phosphocreatine [PCr]) and pH, phosphorus P 31 magnetic resonance spectroscopy ((31)P MRS) provides a unique, noninvasive method to investigate this hypothesis further. METHODS: Calf muscle bioenergetics were studied in 12 men with moderate claudication (ankle-brachial index >/=0.5 and .5, Pearson moment correlation). CONCLUSIONS: Phosphorus 31 MRS provides the first direct evidence of defective energy metabolism in the mitochondria of claudicating calf muscle. This defect appears to be independent of both arterial flow and the severity of occlusive disease in patients with mild to moderate claudication. Coupled with documented ultrastructural and DNA abnormalities in the mitochondria of claudicating skeletal muscle, these data provide evidence for a secondary cause of muscle dysfunction in intermittent claudication.

31P MRS studies of exercising human muscle at high temporal resolution.

Methods for measuring mitochondrial activity from 31P magnetic resonance spectroscopy data collected during and after exercise were compared in controls, weight lifters, and peripheral vascular occlusive disease (PVOD) patients. There were trends toward increasing mitochondrial activity during exercise in order from PVOD patients, moderately active controls, highly active controls, to weight lifters. Results from PVOD patients show divergence of some measures due to 1) the non-exponential nature of phosphocreatine recovery, and 2) potential breakdown of [ADP] control of the mitochondria due to lack of oxygen (for Qmax calculation). These results demonstrate the utility of obtaining and directly analyzing high time resolution data rather than assuming monoexponential behavior of metabolite recovery.

Response to angiotensin inhibition in rats with sustained renovascular hypertension correlates with response to removing renal artery stenosis.

PURPOSE: Sustained (late-phase) renovascular hypertension is associated with lower plasma renin activity than is the early phase. It is not clear to what extent this reduced plasma renin activity reflects diminished influence of the renin-angiotensin system. It also is not clear whether this change in the character of the disease influences the effectiveness of surgical removal of the renal artery stenosis in reversing hypertension. Using an animal model of sustained (> or =10 weeks after renal artery clipping) two-kidney, one-clip renovascular hypertension, we hypothesized that the magnitude of the depressor response to selective angiotensin II receptor blockade with losartan would reflect the influence of the renin-angiotensin system on hypertension and enable us to predict the depressor response to subsequent surgical removal of the clip. METHODS: The left renal arteries of 20 male Sprague-Dawley rats weighing 150 to 200 gm were fitted with a silver clip (0.23 mm internal diameter). Systolic blood pressure was measured by means of tail-cuff plethysmography for 10 weeks. Rats were then given losartan orally (30 mg/kg a day) for 1 week while blood pressure was monitored. After an additional week to allow recovery, 13 rats underwent surgical unclipping, and seven underwent sham repair. Blood pressure again was monitored over the final week. RESULTS: All two-kidney one-clip rats had hypertension 10 weeks after clipping (mean systolic blood pressure 206 +/- 10 mm Hg). Losartan decreased systolic blood pressure by 36 +/- 6 mm Hg. The response was variable, ranging from 3 to 66 mm Hg, and overall blood pressure did not normalize (170 +/- 8 mm Hg). Subsequent surgical unclipping decreased systolic blood pressure by 46 +/- 9 mm Hg. Again the response was variable, ranging from 10 to 99 mm Hg, although overall blood pressure did not normalize (164 +/- 7 mm Hg). The decrease in blood pressure after unclipping showed a high correlation with the blood pressure decrease after losartan administration (r = 0.861, p < 0.001). Resting plasma renin activity (before intervention) was 16 +/- 4 ng angiotensin I per milliliter per hour and was not predictive of the response to either losartan or surgical unclipping. The rats subjected to sham operations had no statistically significant changes in blood pressure. Histologic evaluation showed patent renal arteries without appreciable stenosis or intimal hyperplasia after removal of the clips. CONCLUSIONS: In sustained two-kidney, one-clip renovascular hypertension, the depressor response to angiotensin II receptor blockade is attenuated, suggesting that late-phase hypertension becomes increasingly angiotensin II-independent. In our model, the extent to which sustained renovascular hypertension becomes refractory to 7 days of angiotensin II blockade is highly predictive of the ultimate outcome of surgical repair of renal artery stenosis.

Superficial temporal artery aneurysms.

OBJECTIVE: We analyzed the data from our vascular registry to determine the cause, clinical features, and cost-effective management of this uncommon pathologic entity. DESIGN: Patients referred to the vascular surgery outpatient clinic of a tertiary referral center during the past 18 years were evaluated. SUBJECTS: The subjects were six male patients (14 to 32 years) referred for evaluation of a unilateral pulsatile mass over the temporal region of the head. INTERVENTION: Diagnosis of superficial temporal artery aneurysm was verified by loss of the aneurysm's pulse with compression of the ipsilateral proximal superficial temporal artery. All treated aneurysms were electively ligated and excised as an ambulatory procedure. RESULTS: The symptoms were resolved. No recurrences or other complications were seen. CONCLUSIONS: Although rare, a superficial temporal artery aneurysm should be considered when a temporal head mass is evaluated. This condition is almost always a result of blunt or penetrating head trauma. Clinical examination is sufficient to confirm the diagnosis. Simple elective ligation and excision of the aneurysm is curative.

Secondary aortoesophageal fistula.

Secondary aortoesophageal fistula is the rarest type of aortopeptic fistula, characterized by communication between the reconstructed aorta and the esophagus. This condition has been reported to be uniformly fatal, even after prompt diagnosis and treatment. We report what may be the first case of a successfully managed secondary aortoesophageal fistula that occurred 14 months after repair of a Crawford type II thoracoabdominal aortic aneurysm. The entity was diagnosed with a combination of esophagogastroduodenoscopy and aortography. At exploration, a communication between the proximal anastomosis and the esophagus with otherwise minimal mediastinal contamination was encountered. The anastomosis was replaced with an interposition polytetrafluoroethylene graft, and the esophageal defect was debrided, primarily closed, and reinforced with adjacent old aneurysm wall. There were no postoperative complications, and the patient remains well 18 months after fistula repair.