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BACKGROUND Although the combination of doxorubicin (DOX) and trastuzumab (TRZ) reduces the progression and recurrence of breast cancer, these anticancer drugs are associated with significant cardiotoxic side effects. OBJECTIVE We investigated whether prophylactic administration of flaxseed (FLX) and its bioactive components, α-linolenic acid (ALA) and secoisolariciresinol diglucoside (SDG), would be cardioprotective against DOX + TRZ-mediated cardiotoxicity in a chronic in vivo female murine model. METHODS Wild-type C57BL/6 female mice (10-12 wk old) received daily prophylactic treatment with one of the following diets: 1) regular control (RC) semi-purified diet; 2) 10% FLX diet; 3) 4.4% ALA diet; or 4) 0.44% SDG diet for a total of 6 wks. Within each arm, mice received 3 weekly injections of 0.9% saline or a combination of DOX [8 mg/(kg.wk)] and TRZ [3 mg/(kg.wk)] starting at the end of week 3. The main outcome was to evaluate the effects of FLX, ALA, and SDG on cardiovascular remodeling and markers of apoptosis, inflammation, and mitochondrial dysfunction. Significance between measurements was determined using a 4 (diet) × 2 (chemotherapy) × 2 (time) mixed factorial design with repeated measures. RESULTS In the RC + DOX + TRZ-treated mice at week 6 of the study, the left ventricular ejection fraction (LVEF) decreased by 50% compared with the baseline LVEF (P

Although anticancer systemic therapy agents clearly lead to improved survival in patients with cancer, these can come at the cost of serious complications including cardiotoxicity. Two types of targeted systemic therapies currently in use for colorectal cancer (CRC) and renal cell cancer (RCC), respectively, include the vascular endothelial growth factor inhibitor bevacizumab (BVZ) and the tyrosine kinase inhibitor sunitinib (SNT). Despite the beneficial effects of BVZ and SNT in improving clinical outcomes in the settings of CRC and RCC, there is an increased risk of cardiac dysfunction. The aim of the present study was to determine whether prophylactic administration of renin-angiotensin system (RAS) inhibitors would attenuate the cardiotoxic side effects of BVZ or SNT in a chronic in vivo murine model. A total of 194 wild-type C57Bl/6 male mice received: 1) 0.9% saline, 2) BVZ (10 mg·kg−1·wk−1), or 3) SNT (40 mg·kg−1·day−1) for 4 wk. Within each arm, mice received daily prophylactic treatment with hydralazine (0.05 mg/ml), aliskiren (50 mg/kg), perindopril (4 mg/kg), or valsartan (2 mg/kg). Although hydralazine effectively lowered blood pressure in BVZ- or SNT-treated mice, it did not prevent left ventricular systolic dysfunction. Prophylactic administration of aliskiren, perindopril, or valsartan prevented adverse cardiovascular remodeling in mice treated with either BVZ or SNT. The addition of RAS antagonists also downregulated expression of phosphorylated p38 and Bcl-2-like 19-kDa interacting protein 3 in SNT-treated mice. In our chronic in vivo murine model, RAS antagonists partially attenuated the development of BVZ- or SNT-mediated cardiac dysfunction. Future clinical studies are warranted to investigate the cardioprotective effects of prophylactic treatment with RAS inhibitors in the settings of CRC and RCC. NEW & NOTEWORTHY In the evolving field of cardio-oncology, bevacizumab and sunitinib improve clinical outcomes in the settings of metastatic colorectal cancer and renal cell cancer, respectively. These anticancer drugs, however, are associated with an increased risk of cardiotoxicity. The prophylactic administration of renin-angiotensin system antagonists is partially cardioprotective against bevacizumab- and sunitinib-mediated cardiac dysfunction.

Background Normothermic ex vivo heart perfusion (EVHP) has been shown to improve the preservation of hearts donated after circulatory arrest and to facilitate clinical successful transplantation. Steroids are added to the perfusate solution in current clinical EVHP protocols; however, the impact of this approach on donor heart preservation has not been previously investigated. We sought to determine the impact of steroids on the inflammatory response and development of myocardial edema during EVHP. Methods Thirteen pigs were anesthetized, mechanical ventilation was discontinued, and a hypoxemic cardiac arrest ensued. A 15-minute warm-ischemic standoff period was observed, and then hearts were resuscitated with a cardioplegic solution. Donor hearts were then perfused ex vivo in a normothermic beating state for 6 hours with 500 mg of methylprednisolone (steroid: n = 5) or without (control: n = 8). Results The addition of steroids to the perfusate solution reduced the generation of proinflammatory cytokines (interleukin-6, -8, -1β, and tumor necrosis factor-α) and the development of myocardial edema during EVHP (percentage of weight gain: control = 26% ± 7% versus steroid = 16% ± 10%, p = 0.049). Electron microscopy suggested less endothelial cell edema in the steroid group (injury score: control = 1.8 ± 0.2 versus steroid = 1.2 ± 0.2, p = 0.06), whereas perfusate troponin-I (control = 11.9 ± 1.9 ng/mL versus steroid = 9.5 ± 2.4 ng/mL, p = 0.448) and myocardial function were comparable between the groups. Conclusions The addition of methylprednisolone to the perfusion solution minimizes the generation of proinflammatory cytokines and development of myocardial edema during normothermic ex vivo perfusion of hearts donated after circulatory arrest.

Background In the breast cancer setting, anticancer therapies including doxorubicin (DOX) and trastuzumab (TRZ) are associated with a significantly increased risk of cardiotoxicity. Despite the increasing support for the role of oxidative stress (OS) in its pathophysiology, we still do not have an optimal antioxidant for the prevention of DOX + TRZ–mediated cardiac dysfunction. The objective of this study was to investigate whether the novel antioxidant N-acetylcysteine amide (NACA) can attenuate DOX + TRZ–induced heart failure in a murine model. Methods A total of 100 C57Bl/6 female mice received 1 of the following drug regimens: (1) saline, (2) NACA, (3) DOX, (4) TRZ, (5) DOX + TRZ, (6) NACA + DOX, (7) NACA + TRZ, and (8) NACA + DOX + TRZ. Serial echocardiography was performed over a 10-day study period, after which the mice were killed for histologic and biochemical analyses. Results In mice receiving DOX, the left ventricular ejection fraction (LVEF) decreased from 73% ± 4% to 43% ± 2% on day 10. In mice receiving DOX + TRZ, the LVEF decreased from 72% ± 3% to 32% ± 2% on day 10. Prophylactic administration of NACA to mice receiving DOX or DOX + TRZ was cardioprotective, with an LVEF of 62% ± 3% and 55% ± 3% on day 10, respectively. Histologic and biochemical analyses demonstrated a loss of cellular integrity, increased OS, and increased cardiac apoptosis in mice treated with DOX + TRZ, which was attenuated by the prophylactic administration of NACA. Conclusions NACA attenuated the cardiotoxic side effects of DOX + TRZ in a murine model of chemotherapy-induced cardiac dysfunction by decreasing OS and apoptosis.

The resuscitation of hearts donated after circulatory death (DCD) is gaining widespread interest; however, the method of initial reperfusion (IR) that optimizes functional recovery has not been elucidated. We sought to determine the impact of IR temperature on the recovery of myocardial function during ex vivo heart perfusion (EVHP). Eighteen pigs were anesthetized, mechanical ventilation was discontinued, and cardiac arrest ensued. A 15-min standoff period was observed and then hearts were reperfused for 3 min at three different temperatures (5°C; N = 6, 25°C; N = 5, and 35°C; N = 7) with a normokalemic adenosine-lidocaine crystalloid cardioplegia. Hearts then underwent normothermic EVHP for 6 h during which time myocardial function was assessed in a working mode. We found that IR coronary blood flow differed among treatment groups (5°C = 483 ± 53, 25°C = 722 ± 60, 35°C = 906 ± 36 mL/min, p

Patients with glioblastoma multiforme (GBM) have an average life expectancy of approximately 15 months. Recently, statins have emerged as a potential adjuvant cancer therapy due to their ability to inhibit cell proliferation and induce apoptosis in many types of cancer. The exact mechanisms that mediate the inhibitory actions of statins in cancer cells are largely unknown. The purpose of this proceeding paper is to discuss some of the known anticancer effects of statins, while focusing on GBM therapy that includes adjunct therapy of statins with chemotherapeutic agents.

The recent introduction of novel anticancer therapies, including bevacizumab (BVZ) and sunitinib (SNT), is associated with an increased risk of cardiotoxicity. However, early identification of left ventricular (LV) systolic dysfunction may facilitate dose modification and avoid the development of advanced heart failure. Using a murine model of BVZ- and SNT-mediated cardiotoxicity, we investigated whether cardiac biomarkers and/or tissue velocity imaging (TVI) using echocardiography can detect early changes in cardiac function, before a decrease in LV ejection fraction is identified. A total of 75 wild-type C57Bl/6 male mice were treated with either 0.9% saline, BVZ, or SNT. Serial monitoring of blood pressure, high-sensitivity troponin I, and echocardiographic indexes were performed over a 14-day study period, after which the mice were euthanized for histological and biochemical analyses. Mice treated with either BVZ or SNT developed systemic hypertension as early as day 7, which increased by day 14. Cardiac biomarkers, specifically high-sensitivity troponin I, were not predictive of early LV systolic dysfunction. Although conventional LV ejection fraction values decreased at day 13 in mice treated with either BVZ or SNT, TVI confirmed early LV systolic dysfunction at day 8. Histological and biochemical analysis demonstrated loss of cellular integrity, increased oxidative stress, and increased cardiac apoptosis in mice treated with BVZ or SNT therapy at day 14. In a murine model of BVZ- or SNT-mediated cardiomyopathy, noninvasive assessment by TVI detected early LV systolic dysfunction before alterations in conventional echocardiographic indexes.

Doxorubicin (DOX) and trastuzumab (TRZ) are highly effective chemotherapeutic agents in the breast cancer setting, limited by their cardiotoxic side effects. Among the potential mechanisms for this drug-induced cardiomyopathy, increased production of oxidative stress (OS) through a nitric oxide synthase 3 (NOS3)-dependent pathway has gained recent attention. The objective of the study was to determine the role of NOS3 and OS in a clinically relevant female murine model of DOX- and TRZ-induced heart failure.A total of 120 female mice (60 wild-type [WT] and 60 NOS3 knockout [NOS3(-/-)]) were treated with either 0.9% saline, DOX, TRZ, or DOX with TRZ (DOX+TRZ). Serial echocardiography was performed for a total of 10 days, after which the mice were euthanized for histological and biochemical analyses.In WT female mice receiving DOX+TRZ, left ventricular ejection fraction (LVEF) decreased from 75 ± 3% at baseline to 46 ± 2% at day 10 (P0.05). In the NOS3(-/-) group, LVEF decreased from 72 ± 3% at baseline to 35 ± 2% at day 10 (P0.05). LVEF was significantly lower in NOS3(-/-) female mice receiving DOX+TRZ than WT mice at day 10 (P0.05). Compared with WT, NOS3(-/-) female mice also demonstrated increased mortality after treatment with DOX+TRZ, corroborating the echocardiographic findings. Histological analysis demonstrated increased myofibrillar degradation and loss of cell integrity in NOS3(-/-) female mice treated with DOX+TRZ. There was increased generation of oxidized phosphatidylcholine, a marker of OS, in NOS3(-/-) female mice receiving DOX+TRZ compared with control mice.Congenital absence of NOS3 potentiates the cardiotoxic side effects of DOX+TRZ in an acute female murine model of chemotherapy-induced cardiomyopathy.

Glioblastoma multiforme (GBM) is the deadliest brain tumor with an approximate 14 month survival rate after diagnosis and treatment. Temozolomide (TMZ), the chemotherapeutic drug of choice for GBM, is an alkylating agent that causes DNA damage. TMZ treatment results in the induction of apoptosis in GBM cells, however, it induces autophagy and consequently chemoresistance. Statins are mevalonate (MEV) cascade inhibitors with beneficial effects on the enhancement of the survival rate of patients with different types of cancer. Here, we determined the effect of simvastatin (Simva), a blood brain barrier permeable statin, on the sensitization of GBM cells to TMZ induced apoptosis through inhibition of autophagy flux. We pretreated two GMB cell lines, U251 and U87 cells, with low doses of Simva (1 and 2.5 M, respectively) with or without different intermediates of the mevalonate cascade and then treated cells with TMZ (100 M) for 48-96 hrs. A signficiantly reduced viability and increased in the population of apoptotic dead cells were observed in GBM cells treated with the Simva-TMZ compared to cells treated with TMZ alone. Addition of MEV, Farnesyl pyrophosphate, Geranylgeranyl pyrophosphate and cholesterol did not attenuate these effects significantly. Sima-TMZ treatment did not alter the total cholesterol pool in U87 and U251 cells compared to controls. Western blot analysis, immunocytochemistry and transmission electron microscopy revealed that Simva-TMZ inhibited autophagic flux. Overall, the results suggest that sensitization of GBM cells to TMZ-induced apoptosis by Simva is independent on the cholesterol biosynthetic pathway but may involve inhibition of autophagy.

Background: Hearts donated after circulatory death may represent an additional donor source. The influx of sodium and calcium ions across the sarcolemma play a central role in the pathogenesis of ischemia-reperfusion injury; however, this process may be inhibited if the initial reperfusion solution is rendered hypocalcemic and acidic. We sought to determine the calcium concentration and pH of the initial reperfusion solution that yielded optimal functional recovery of hearts donated after circulatory death during ex vivo heart perfusion. Methods: Pigs were anesthetized, mechanical ventilation was discontinued, and a 15-minute standoff period was observed after circulatory arrest. Hearts were reperfused with a normothermic cardioplegia of varying calcium concentrations (part 1 [50 μmol/L, n = 4; 220 μmol/L, n = 9; 500 μmol/L, n = 4; and 1,250 μmol/L, n = 5]) and pH (part 2 [7.9, n = 5; 7.4, n = 9; 6.9, n = 8; and 6.4, n = 6]). Myocardial function was then assessed in a physiologic working model 1 hour after initiation of normothermic ex vivo heart perfusion. Results: The calcium concentration and pH of the cardioplegic solution affected the development of myocardial edema (part 1: 50 μmol/L = 5.8% ± 0.9%; 220 μmol/L = 4.3% ± 0.4%; 500 μmol/L = 7.0% ± 0.6%; and 1,250 μmol/L = 6.6% ± 0.8% weight gain, p = 0.015; part 2: 7.9 = 3.6% ± 0.4%, 7.4 = 4.3% ± 0.4%, 6.9 = 3.7% ± 0.6%, and 6.4 = 6.4% ± 1.3% weight gain, p = 0.056) and the recovery of myocardial function (cardiac index part 1: 50 μmol/L = 2.6 ± 0.6; 220 μmol/L = 6.0 ± 0.8; 500 μmol/L = 2.3 ± 0.5; and 1,250 μmol/L = 1.9 ± 0.6 mL · m−1 · g−1, p < 0.001; part 2: 7.9 = 1.5 ± 0.7; 7.4 = 6.0 ± 0.8; 6.9 = 8.4 ± 1.8; and 6.4 = 3.1 ± 0.8 mL · m−1 · g−1, p = 0.003) during ex vivo heart perfusion. Conclusions: Initial reperfusion of hearts donated after circulatory death with a hypocalcemic and moderately acidic cardioplegia minimizes edema and optimizes functional recovery during subsequent ex vivo heart perfusion.

Vanadium can induce potent hypoglycemic effects in type 1 and type 2 diabetes mellitus animals, but toxic adverse effects have inhibited the translation of these findings. Administration of vanadate in a black tea decoction has shown impressive hypoglycemic effects without evidence of toxicity in short-term studies. The purpose of this study was to investigate the hypoglycemic action and the toxic adverse effects of a tea/vanadate (T/V) decoction in diabetic rats over a 14-month treatment period. Streptozotocin-induced type 1 diabetes mellitus rats were orally gavaged with 40 mg sodium vanadate in a black tea decoction only when blood glucose levels were greater than 10 mmol/L. Glycemic status and liver and kidney function were monitored over 14 months. All of the diabetic rats in this treatment group (n = 25) required treatment with the T/V decoction at the start of the study to reduce blood glucose levels to less than 10 mmol/L. Diarrhea was uncommon among the T/V-treated animals during the first week of T/V treatment and was absent thereafter. There was no evidence of liver or kidney dysfunction or injury. From 2 to 6 months, fewer animals required the T/V treatment to maintain their blood glucose levels. After 9 months of treatment, none of the diabetic animals required any T/V to maintain their blood glucose levels at less than 10 mmol/L. Oral administration of a T/V decoction provides safe, long-acting hypoglycemic effects in type 1 diabetes mellitus rats. The typical glycemic signs of diabetes were absent for the last 5 months of the study.

Objective Although the combination of doxorubicin (Dox) and trastuzumab (Trz) reduces breast cancer progression and recurrence, it is limited by significant cardiotoxic side effects. Little is known about the utility of antioxidants in the prevention of this drug-induced cardiomyopathy. The aim of the study was to determine whether the antioxidant probucol (Prob) would be useful in attenuating Dox and Trz-mediated cardiotoxicity. Methods A total of 114 mice were randomized to treatment with Trz, Dox, or Dox+Trz. Within each arm, mice received prophylactic treatment with placebo or Prob. Serial murine echocardiography with tissue Doppler imaging was performed daily for 10 days. At 10 days posttreatment, the hearts were removed for histopathologic and Western blot analyses. Results Left ventricular cavity dimensions and systolic parameters were preserved in mice prophylactically treated with Prob after the administration of Dox+Trz. Although the combination of Dox+Trz demonstrated >80% mortality at day 5, prophylactic treatment with Prob reduced mortality to 40% at day 10. There was decreased histologic evidence of cardiac damage and reduced apoptosis due to Dox+Trz in mice pretreated with Prob. Conclusion The cardiotoxic effects of Dox+Trz are partially attenuated by the prophylactic administration of the antioxidant Prob.

Lung cancer is the leading cause of death worldwide. Physical and chemical agents such as tobacco smoke are the leading cause of various lung cancers. The intrinsic heterogeneity of normal lung tissue may be affected in different ways, giving rise to different types of lung cancers classified as either small‐cell lung cancer (SCLC) or non‐small cell lung cancer (NSCLC). Adenocarcinoma, a NSCLC, accounts for 40 percent of all lung cancer cases and the incidence is increasing worldwide, especially among women. The survival rate and prognosis is poorest for adenocarcinoma. Therefore, diagnosis at the earliest stage (Stage I, localized) is critical for increasing survival rates of those suffering from lung cancer. However, many factors affect early diagnosis including the variable natural growth of tumors plus technological and human factors associated with manipulation of tissue samples and interpretation of results. This article reviews potential problems associated with diagnosing lung cancer and considers future directions of diagnostic technology.

s S251 normothermic beating state and transitioned into working mode for assessment of myocardial function by measuring the cardiac index (mL/minute/gram heart tissue) achieved at a left atrial pressure of 8 mmHg and an aortic diastolic pressure of 40 mmHg. RESULTS: Hearts sustained an equivalent period of warm ischemia (5 C1⁄428 1, 25 C1⁄429 1, 35 C1⁄427 1 minutes, p1⁄40.50) prior to IR. During IR coronary blood flow (5 C1⁄4483 53, 25 C1⁄4722 60, 35 C1⁄4906 36 mL/min, p

We have previously reported that weekly administration of the adenosine deaminase inhibitor, 2'-deoxycoformycin (dCF), reduces the incidence of insulin-dependent diabetes mellitus (IDDM) in the BB Wistar rat, and this effect is likely due to immunosuppression by dCF. In the present study, we examined the effect of altering the dose and scheduling of dCF on prevention of IDDM in the BB rat. When rats were treated from day 25 of age with 2.5, 4, or 10 mg of dCF/kg/week, the percentage of diabetes-free animals at 120 days of age was 40, 60, and 80% respectively, compared with 10% for control animals, demonstrating increased protection against IDDM with increased dCF dose. Histological assessment of the pancreata from animals that became diabetic revealed a marked mononuclear infiltrate and a loss of positive staining for beta cell granules. In contrast, pancreata from animals that remained diabetes-free appeared normal. Protection against IDDM by dCF was time dependent and only occurred if treatments were initiated by day 30 of age. In addition, the protective effect persisted after drug withdrawal. Further studies are required to determine the optimum duration of therapy with dCF to prevent IDDM and to examine the immunological mechanism responsible for this effect.

Background Cigarette smoking is the leading cause of preventable death and has been implicated in pathogenesis of pulmonary, oral and systemic diseases. Smoking during pregnancy is a risk factor for the developing fetus and may be a major cause of infant mortality. Moreover, the oral cavity, and all cells within are the first to be exposed to cigarette smoke and may be a possible source for the spread of toxins to other organs of the body. Fibroblasts in general are morphologically heterogeneous connective tissue cells with diverse functions. Apoptosis or programmed cell death is a crucial process during embryogenesis and for the maintenance of homeostasis throughout life. Deregulation of apoptosis has been implicated in abnormal lung development in the fetus and disease progression in adults. Caspases are proteases which belong to the family of cysteine aspartic acid proteases and are key components for downstream amplification of intracellular apoptotic signals. Of 14 known caspases, caspase-3 is the key executioner of apoptosis. In the present study we explored the hypothesis that cigarette smoke (CS) extract activates caspase-3 in two types of fibroblasts, both of which would be exposed directly to cigarette smoke, isolated fetal rat lung fibroblasts and adult rat periodontal ligament (PDL) fibroblasts. Methods Isolated fetal rat lung fibroblasts and adult PDLs were used. Cells were exposed to different concentrations of CS for 60 min. Caspase-3 activity and its inhibition by Z-VAD-fmk were measured by caspase-3 fluorometric assay. The effect of CSE on cellular viability was measured using the MTT formazan assay. Caspase-3 expression was detected by western blot analysis and cellular localization of caspase-3 was determined by immunofluorescence using fluorescence microscopy. Results It was observed in fetal rat lung fibroblast cells that CSE extract significantly (p

Thermal infrared spectroscopy has become an increasingly important tool for remote compositional analysis and geologic mapping. Most published laboratory measurements have been obtained in bidirectional reflection or transmission, whereas remotely sensed thermal infrared data are obtained by measuring the emitted energy. Section 2 of this paper describes a laboratory technique for determining calibrated emissivities of natural surfaces. Equations are developed to account for the energy reflected from the environment and to determine directly the sample temperature from measurements of hot and cold blackbody targets. Two methods for determining emissivity are developed: one in which only a hot sample measurement is made and the reflected background energy is removed by modeling, and a second in which the sample is cooled and the reflected energy is measured directly. Relative emissivity can be obtained to approximately 1% and absolute emissivities can be obtained to 2-15%, depending on the validity of the assumption that the emissivity of the sample is unity at some wavelength. The emission data agree well within the hemispherically integrated reflection data but point out probelms associated with bidirectional reflectance measurements. Section 3 applies emissivity measurements to the study of layered surfaces consisting of desert varnish coatings on granite and granodiorite rock suites. Two linear models are developed: the first assumes linear mixing of independent emission from the substrate and varnish (checkerboard model); the second models tansmission through an absorbing/emitting medium. Regardless of whether the varnish is or is not relatively transparant and strongly absorptive, the spectral effect of varnish increases linearly with varnish thickness, indicating that thick patches of varnish dominate the spectral properties. As a result, medium varnish thickness can be determined from spectral measurements. In addition, the composition of a substrate can be estimated through varnish layers up to 40-50 micrometers in median thickness, and the composition of the varnish material can be determined if the substrate material is known. The varnishes studied are composed primarily of clay materials, consistent with previous studies of varnish composition.

The effect of the adenosine deaminase (ADA) inhibitor 2′-deoxycoformycin (dCF) on the development of insulin-dependent diabetes mellitus (IDDM) was assessed in the BB Wistar rat. Sixty-one male rats were treated from days 30 to 120 with 0, 0.5, 1.0 or 1.5 mg dCF/kg/week. The incidence of IDDM was 78% in the controls and was significantly (P < 0.01) decreased in rats receiving 1.5 mg dCF/kg/week (32%), but not in rats receiving lower doses of the drug. However, for those rats that became diabetic the mean time to the development of IDDM was unchanged in animals receiving dCF compared with control. dCF treatment did not produce significant weight loss in the animals or gross changes in the thymus, spleen or kidneys. Although the protective effect of dCF against IDDM was likely produced by immunosuppression, the different dCF dosages had similar effects on ADA suppression in spleen or thymus and on dATP accumulation in these organs.

Heterotopic heart transplants were performed on 50 New Zealand white rabbits. Groups of 5 rabbits were randomly assigned to receive, through an intravenous route, rapamycin (RAPA) or cyclosporine at the following doses: RAPA (0.05, 0.1, 0.5, and 1.0 mg/kg/day); CsA (5.0, 10.0, and 15.0 mg/kg/day). Drug vehicle and saline controls were also included. Trough blood concentrations were monitored in both RAPA- and CsA-treated groups on a weekly basis throughout the study. Biochemical assessment of renal and liver function was performed at the beginning and end of the study. Animals receiving RAPA exhibited excellent allograft survival; only two animals in the lowest dosage group (0.05 mg/kg/day) rejected their grafts. In contrast, no rejection occurred in the CsA-treated groups. Animals that rejected their grafts were maintained on the drug until the endpoint of the study was reached at 60 days posttransplant to monitor drug induced side-effects. In some instances animals were sacrificed prior to this time due to infectious and other complications. No significant changes in renal or liver function were noted in the RAPA-treated group, while in the group of animals receiving the highest dose of CsA (15.0 mg/kg/day) a significant decrease in creatinine clearance was noted. A correlation was shown to exist between dose and the trough concentrations of both drugs. The whole-blood concentrations of RAPA that resulted in maximal efficacy with minimal toxicity was in the range of 10-60 micrograms/L. Rabbits having trough whole-blood concentrations of < 10 micrograms/L rejected their grafts. A much wider therapeutic range for CsA (50-300 micrograms/L) was noted. The results suggest that RAPA is as efficacious as CsA in prevention of allograft rejection in the animal model tested. The therapeutic monitoring of trough blood concentrations of RAPA, as with CsA, may be useful in guiding dosage adjustments to maximize the immunosuppressive efficacy while minimizing drug-induced side-effects.

s S251 normothermic beating state and transitioned into working mode for assessment of myocardial function by measuring the cardiac index (mL/minute/gram heart tissue) achieved at a left atrial pressure of 8 mmHg and an aortic diastolic pressure of 40 mmHg. RESULTS: Hearts sustained an equivalent period of warm ischemia (5 C1⁄428 1, 25 C1⁄429 1, 35 C1⁄427 1 minutes, p1⁄40.50) prior to IR. During IR coronary blood flow (5 C1⁄4483 53, 25 C1⁄4722 60, 35 C1⁄4906 36 mL/min, p

Objective To describe the clinical courses of patients who received intravitreal injections of highly concentrated amphotericin B deoxycholate for suspected fungal endophthalmitis. Methods Retrospective medical record review of 3 cases of intraocular toxicity from highly concentrated amphotericin B. Results The first patient developed posttraumatic endophthalmitis and received an undiluted dose (500 μg) of amphotericin B. He developed severe intraocular inflammation and required a pars plana lensectomy, vitrectomy, and scleral buckle after developing a cataract and retinal detachment. Six years later, his visual acuity stabilized at 20/30. The second patient developed endogenous endophthalmitis and was treated with 5 intravitreal injections of amphotericin B and underwent 3 surgical procedures. The surgeon later discovered that the patient had received 55 μg of amphotericin B during the second injection. Three months after the injection, the patient's visual acuity was 20/60. The third patient developed chronic postoperative endophthalmitis following cataract extraction. He received 160 μg of amphotericin B and was immediately treated with a vitreous washout. Two years later, his visual acuity improved to 20/30. The vitreous culture results were negative in each case. A key finding was that the amphotericin B solution appeared to be yellow instead of nearly colorless. Conclusions We present 3 cases of intraocular toxicity from highly concentrated amphotericin B. In every case, the overly concentrated amphotericin B solution was yellow in color. Although severe noninfectious panophthalmitis resulted in every case, the visual acuity outcomes were good. Physicians should examine the color of amphotericin B solution prior to intraocular administration. If the solution appears to be yellow, the medication should not be injected.

Ciclosporin (CsA) was administered intravenously for 30 days to New Zealand white rabbits at the following doses: group A, 2.5 mg/kg/day; group B, 5.0 mg/kg/day; group C, 10 mg/kg/day; group D, saline control; group E, vehicle (cremophor-EL) control. Creatinine clearance was significantly reduced (p less than 0.05) at time of sacrifice in group C as compared to controls. There were no apparent differences among the five groups with respect to body weight gain or blood pressure. Morphologically, there were marked changes in the cytoarchitecture of the kidneys from all groups of animals treated with CsA. At the light microscopic level, in contrast to controls, there was the presence of leukocyte infiltration, tubular atrophy, interstitial fibrosis, and arteriolopathy. At the ultrastructural level, numerous vacuoles, lysosomal-like structures, and loss of cellular integrity were seen in the proximal and distal tubules, as well as interstitial fibrosis in the form of numerous collagen fibers. No changes in the glomeruli were seen in any of the CsA-treated groups. These findings are consistent with chronic CsA nephrotoxicity similar to that seen in man.

It has been established that phospholipids and cholesterol interact in films of pulmonary surfactant (PS). Generally it is thought that phospholipids increase film stability whereas cholesterol increases film fluidity. To study this further, we modified dietary cholesterol in mice which received either standard rodent lacking cholesterol (sd), or high cholesterol (2%) diet (hc) for 1 month. Phospholipid stability was investigated by a capillary surfactometer (CS), which measures airflow resistance and patency. PS was collected by bronchiolar lavage and centrifuged to obtain the surface-active film (SAF). Results showed that the hc-SAF had significantly more cholesterol than sd-SAF. CS analyses at 37°C showed no significance differences in airflow resistance between hc-SAF and sd-SAF. However, at 37°C, sd-SAF showed greater ability to maintain patency compared to hc-SAF, whereas at 42°C hc-SAF showed patency ability similar to sd-SAF. The results suggested that increased cholesterol in hc-SAF induced less stability in the SAF possibly due to cholesterol's fluidizing effect on phospholipids at physiological temperatures. Pediatr Pulmonol. 2008; 43:426–434. © 2008 Wiley-Liss, Inc.

This research was designed to evaluate musculoskeletal anatomy of the quadriceps region relative to the patellofemoral joint. The hypothesis for the study was that the oblique portion (VMO) of the vastus medialis muscle (VM) is anatomically positioned to function primarily as an active medial stabilizer of the patella. Because many clinicians believe that the VMO functions independently as an active medial stabilizer of the patellofemoral joint (PFJ), PFJ rehabilitation protocols commonly target the VMO in an attempt to restore normal joint mechanics. It is unclear whether this purported selective function is supported by the underlying anatomical structure. Through dissection of 32 limbs from 24 intact cadavers with normal patellar alignment, data were collected on VM fiber alignment and innervation, the presence of fascial plane, and the length of VM about the patella. Statistical analyses demonstrated that the oblique and long heads of the VM muscle had significantly different (P < 0.05) angles of fiber orientation, as expected. When measurements were taken relative to a vertical axis (standardizing limb alignment between cadavers), the difference in fiber angles between oblique and long heads of the VM was reduced significantly. Additionally

Background. The practical application of sentinel lymph node biopsy in squamous cell carcinoma of the head and neck is restricted by the time sensitivity of blue dye and lack of spatial resolution and nonspecific node enhancement with radiocolloid. This study evaluates the use of magnetic resonance (MR) lymphangiography and carbon dye labeling to circumvent these limitations. Methods. Gadomer/carbon dye mixture was injected into the tongue and stifle of adult swine (n = 4). MR lymphatic mapping was followed by intraoperative mapping with isosulfan blue dye. Sentinel lymph node biopsy and completion node dissection were performed 60 minutes after injection in four nodal basins and at 7 days after injection in eight. Results. The technique was successful in all 12 nodal basins. The sentinel lymph nodes were stained black at the time of the immediate and delayed dissections Conclusions. MR lymphangiography provides temporal and anatomic localization of the sentinel lymph node with a single investigation. Carbon dye is a sensitive and persistent visual marker of MRI-targeted sentinel lymph nodes.

Summary Valproic acid (VPA) is an antiepileptic drug used clinically. Because of its known teratogenic properties VPA is not recommended for women of child bearing age. The present study was designed to assess the effects of VPA on both fetal and maternal organs. Randomized groups of pregnant mice were treated as follows: Group 1 (n = 10) 500 mg/kg VPA/day on gestation days 8-11; Group 2 (n = 10) 600 mg/kg VPA/day on gestation days 8-11; and Group 3 (n = 4) saline-injected controls. On gestation day 18, the pregnant mice were euthanized, fetuses collected and prepared for scanning electron microscopy. In addition, fetal and maternal organs were processed for routine histology, immunohistochemistry for growth factors (TGF alpha, beta-1, beta-2 and EGF) and transmission electron microscopy. Scanning microscopy revealed specific lesions induced by VPA in the fetus, namely spina bifida occulta, exencephaly, and exophthalmia. On the other hand, there were no detectable morphological changes in fetal or maternal organs by routine histology, immunohistochemistry or electron microscopy. The data suggest that the lesions present in the fetus are due to a direct effect by VPA on retinoic acid, a ubiquitous compound that has a role in normal development, rather than the lack of transport of sufficient nutrients to the fetus as a result of placental insufficiency due to VPA-induced toxicity.

Breast cancer is a leading cause of death in women, occurring in one of eight during their lifetime. A comparable cancer in men, prostate cancer, claims the lives of just under 30,000 men in the United States each year with a prevalence of one in six (1). These tumors, along with carcinomas of the lung, thyroid, and kidney have a high propensity to metastasize to bone, which constitutes one of their most serious complications, creates a great challenge for treatment, and often carries a poor prognosis. Metastases are the most frequent bone tumors and cause significant morbidity due to pain, osteolysis, pathological fractures, hypercalcemia, and anemia (2). Established bone metastases are resistant to treatment and current therapeutic approaches such as endocrine therapy, radiation therapy, and chemotherapy are often ineffective (3). This chapter has been written on the premise that a better understanding of the pathophysiology of bone metastasis may provide insight into the design of diagnostic techniques that would help to identify bone metastasis formation at its earliest stages. This may provide an opportunity for early treatment and alleviate the necessity to treat them at a late stage.

Background and Objectives The challenge for implementation of sentinel lymph node biopsy is to develop a reliable minimally invasive technique that identifies all possible sentinel nodes with high temporal and spatial resolution. This study evaluated the use of a magnetic resonance imaging (MRI) contrast agent (USPIO) for preoperative sentinel node detection. Methods Anesthetized pigs received interstitial or intradermal injections of ultra small superparamagnetic of iron oxide (USPIO) (0.2 or 5 mg Fe) in the L/R posterior tongue and stifles (knee) respectively. MRI was done before, during injection and at 0.25, 0.5, 1, 2, 4, 6, 24, and 48 hr after which isosulfan blue sentinel node mapping was done. Results In the tongue, both doses of USPIO identified the sentinel node in the early images. No additional nodes were detected by MR at 24 or 48 hr. In the hind limb, sentinel nodes identified on the early MR images were also identified by the isosulfan blue. In both locations, the higher dose also identified secondary nodes some of which were also identified by the isosulfan blue. All sentinel nodes that were identified by USPIO on MRI were noted to be stained brown at the time of dissection. Conclusions Interstitial MR lymphangiography is a useful technique for the detection of sentinel lymph nodes. This method provides excellent simultaneous temporal and spatial resolution, is minimally invasive, and can be performed preoperatively. J. Surg. Oncol. 2001;78:151–156. © 2001 Wiley-Liss, Inc.

Working with the College of Physicians and Surgeons of Manitoba, and using tonsillectomies as a basis of inquiry, MCHPE examined surgical rates and patterns of practice. This project had three major aims: to review whether current patterns of delivery provide optimal care; to enhance the development of clinical guidelines; and to inform and influence physician practice.Both a population-based method of inquiry (which permits comparisons across population groups) and a provider-based approach (which offers insights into differences in the nature of care offered by different types of hospitals and physicians) were used.Synergies between these two approaches offered useful insight into aspects of quality and efficiency of care.Consistent with other jurisdictions, there was a high degree of variability across regions. However, there were also a number of surprising findings, including high rates of surgery in females, in older children, and among residents of rural areas. Data analysis raised a number of quality-of-care issues related to small caseload volumes, performance of procedures in very young children, and patterns of postoperative care in rural hospitals. The analyses provided impetus for addressing these issues in the guideline and suggested that the target audience for intervention should be rural physicians rather than urban specialists.This project demonstrated that data analysis can provide a powerful adjunct to the development and implementation of clinical practice guidelines.

Ocular toxicity resulting from chemicals in eye drops is common. Preservatives such as benzalkonium chloride (BAC) and related quaternary ammonium salts have been shown to play an important part in these reactions.1 The symptoms can range from mild injection and itching to severe conjunctival reactions. The term pseudopemphigoid has been applied to drug-induced cicatrising conjunctivitis.2–5 We present a patient who developed drug-induced granulomatous cicatrising conjunctivitis secondary to glaucoma drops. A 60-year-old man presented to the eye clinic for ocular irritation. His ocular history was positive for glaucoma, for which he was using latanoprost (Xalatan, Pfizer, New York, New York, USA), and brimonidine 0.2% (a generic preparation). Examination showed severe conjunctival injection bilaterally, …

Pharmacodynamic (PD) monitoring of immunosuppressive drugs provides a novel approach to optimization of drug therapy in transplant recipients. We chose to investigate this using mycophenolic acid (MPA), an immunosuppressive drug that mediates its effect by the inhibition of inosine monophosphate dehydrogenase (IMPDH), a key enzyme in the de novo biosynthesis of purines. A comparison of the relationship between PD versus drug level monitoring was performed using a heterotopic cardiac transplant in New Zealand white rabbits. The animals were divided into four different treatment groups. Control animals were administered the drug vehicle, the treatment groups were administered mycophenolate mofetil (MMF) at doses of 40, 80, and 160 mg/kg/day. Statistically significant (p < 0.05) prolongation of graft survival was obtained at the 160 mg/kg/day dose group. The mean MPA concentration at this dose was approximately 2.5 mg/l, suggesting that this concentration may provide adequate immunosuppression. An increase in IMPDH activity appeared a few days prior to rejection, suggesting that measurement of enzyme activity may have potential for use as a marker of graft rejection. A significant (p < 0.05) relationship exists between MPA concentration and graft survival and the former with dose of MMF. There was a negative correlation (p = 0.17) between MPA concentration and IMPDH activity, while a trend (p = 0.37) to inverse relationship between graft survival and IMPDH activity was found. The data suggests that the measurement of the biological response may provide a useful adjunct to traditional therapeutic drug monitoring (TDM) for optimization of dosing of immunosuppressive drugs.

Objective To investigate whether a dietary supplement of vitamin B 6 could attenuate the elevation of systolic blood pressure (SBP) in Zucker obese or spontaneously hypertensive rats, or rats ingesting sucrose. Methods Zucker obese rats (fa/fa), Sprague-Dawley rats with sucrose-induced elevation of SBP, spontaneously hypertensive rats (SHRs) and their corresponding controls were tested for the effects of vitamin B ingestion in different ways : (1) vitamin B 6 was included as a supplement (five times the normal intake) from the start of the experiment until the development of hypertension ; (2) vitamin B 6 supplement was removed from the diet of Zucker obese and Zucker lean control groups after 16 weeks on the dietary treatments ; and (3) a diet deficient in vitamin B 6 was instituted in SHRs and control Wistar-Kyoto (WKY) rats. The SBP of rats in all groups was monitored in the conscious animal by tail-cuff plethysmography. The effects of the various treatments on the uptake of calcium by caudal artery segments were examined. Results Male Zucker obese rats (fa/fa) of age 6 weeks fed a commercial rat chow developed hypertension in 3-4 weeks, whereas their lean controls (Fa/Fa) did not The inclusion of a vitamin B 6 supplement (five times the normal intake) resulted in a complete attenuation of the hypertension in the obese strain. Removal of the vitamin B 6 supplement from the diet of these obese rats resulted in the retum of hypertension within 2 weeks. Similar changes in SBP were also observed in the Zucker lean controls treated with vitamin B 6 . The ingestion of sucrose by male Sprague-Dawley rats resulted in modest elevation of SBP that was attenuated by the inclusion of the vitamin B 6 supplement in their diet In contrast, there was no response to the inclusion or removal of dietary vitamin B 6 supplement in the SHRs. However, the WKY control rats responded to both these conditions in a similar manner to that seen in the Sprague-Dawley strain. Increased peripheral resistance resulting from increased permeability of vascular smooth muscle plasma membrane to Ca 2+ is thought to be one of the mechanisms of hypertension. Changes in SBP correlated with changes in the uptake of calcium by caudal artery segments in all the groups studied. The Zucker obese and sucrose-induced hypertensive rats have abnormalities in carbohydrate metabolism. The vitamin B 6 supplement decreased the random or fasting blood glucose levels in the Zucker obese and sucrose-fed rats respectively. Conclusion This is the first observation that animal models of hypertension can be classified on the basis of their response to a vitamin B 6 supplement. On this basis, the etiology of hypertension in SHRs is quite distinct from that in Zucker obese rats and in rats ingesting sucrose.

Bilateral testicular tumors (adrenal rests) may occur in untreated or poorly controlled congenital adrenal hyperplasia. This case report describes two unique associated phenomena: (1) psychologic disturbances similar to those seen with exogenous androgen abuse, which resolved with appropriate glucocorticoid suppression of androgen over-production by this abnormal adrenal/adrenal rest tissue; and (2) testicular failure which showed a partial, delayed recovery with corticosteroid therapy. The need for a careful history and biochemical screening for all patients with bilateral testicular tumors is reinforced.

A computer program, MOD (mixed oligo designer), which aids in planning site-directed mutagenesis experiments using highly substituted oligodeoxyribonucleotides (oligos), is described. The program calculates the relationship between the degree of oligo substitution and the mutation frequency, in order to achieve an optimal level of mutagenesis. The program can be used on a wide variety of computers and runs under a number of different operating systems.

Cyclosporine A (CsA) represents one of the more important therapeutic advances in the field of kidney transplantation. However, its effectiveness is limited by serious side effects, most notably nephrotoxicity. Investigation of the mechanisms of CsA-induced renal dysfunction has been hampered by the lack of a suitable experimental model. The majority of studies using the rodent have failed to exhibit all of the structural changes seen in chronic CsA-induced nephrotoxicity reported in man, using pharmacologic doses administered orally, subcutaneously, or intravenously. More recently, studies using the rabbit as an experimental model have demonstrated leucocyte infiltration, tubular atrophy, interstitial fibrosis, and arteriolopathy after therapeutic doses of CsA over 30 days. These changes are similar to those seen in chronic CsA-induced nephrotoxicity in man.