Bëlohlávek, Marek

Abstract Context Humans with obesity and insulin resistance exhibit lipid accumulation in skeletal muscle, but the underlying biological mechanisms responsible for the accumulation of lipid in the muscle of these individuals remain unknown. Objective We investigated how plasma insulin modulates the extraction of circulating triglycerides (TGs) and nonesterified fatty acids (NEFAs) from ingested and endogenous origin in the muscle of lean, insulin-sensitive humans (Lean-IS) and contrasted these responses to those in humans with obesity and insulin resistance (Obese-IR). Methods The studies were performed in a postprandial state associated with steady-state plasma TG concentrations. The arterio-venous blood sampling technique was employed to determine the extraction of circulating lipids across the forearm muscle before and after insulin infusion. We distinguished the kinetics of TGs and NEFAs from ingested origin from those from endogenous origin across muscle by incorporating stable isotope-labeled triolein in the ingested fat. Results Insulin infusion rapidly suppressed the extraction of plasma TGs from endogenous but not ingested origin in the muscle of the Lean-IS, but this response was absent in the muscle of the Obese-IR. Furthermore, in the muscle of the Lean-IS, insulin infusion decreased the extraction of circulating NEFAs from both ingested and endogenous origin; however, this response was absent for NEFAs from ingested origin in the muscle of the Obese-IR subjects. Conclusion Partitioning of circulating lipids away from the skeletal muscle when plasma insulin increases during the postprandial period is impaired in humans with obesity and insulin resistance.

Background: Myocardial reperfusion following prolonged ischemia may paradoxically cause additional injury, including edema and microvascular damage. These structural alterations augment tissue echogenicity measurable by integrated backscatter (IB). We sought to characterize alterations of myocardial IB in an ischemic and then reperfused region of the rat heart. Methods: Myocardial IB in 12 adult male Sprague-Dawley rats was studied at baseline, 60 minutes after left anterior descending coronary artery occlusion, and chronologically after coronary reopening. Short-axis scans of the mid left ventricle were acquired at a frequency of 8 MHz. Mean±SD values of IB were measured in the region of interest at baseline, ischemia, and reperfusion, and assessed by a paired t-test. Results: See figure and table. IB did not significantly change between baseline and ischemia. However, within 1 minute of reperfusion, IB significantly increased, as compared to both baseline and ischemia, and continued to increase until 10 minutes after reperfusion, when a plateau was reached. Conclusions: Myocardial reperfusion following acute prolonged ischemia in a rat heart significantly augments tissue echogenicity. A continuing increase of IB within minutes of the reperfusion onset and the subsequent plateau of elevated IB values suggest progression and completion, respectively, of reperfusion injury, detectable by a high-frequency transducer.

Abstract In collaboration with the American College of Veterinary Pathologists

By binding to a nebulin site on actin in skeletal myofibrils phalloidin (PH) causes a force decrease (phase 1) followed by an increase in force (phase 2). The initial force decrease likely reflects a disturbance in actin‐nebulin interaction. The aim of this work was to determine the role of myosin cross‐bridges (XBs) and Ca 2+ in PH‐induced force decrease in skinned rabbit psoas fibers. Force response to PH was studied in the presence of 30 mM 2,3‐Butanedione monoxime (an agent reducing a fraction of strongly bound XBs), where we found that the phase 2 response was suppressed and PH induced 60–70% force decrease. Pretreatment with PH of relaxed (pCa 9, strongly bound XBs are absent) or rigor (in the absence of ATP, all XBs are strongly bound to actin) muscles also resulted in force loss. At the same duration of pretreatment, the remaining force was higher and more sensitive to PH in the latter case. The same properties were found for dethiophalloidin (DTPH), where it was also shown that the force loss was essentially the same at pCa 4 and pCa 9. We concluded that rigor XBs, but not Ca 2+ , affect the rate of PH binding to actin in myofibrils. The XBs mediated changes in nebulin site on actin are supportive for suggested regulatory nebulin role. Supported by NIH grants HL68555 and HL37680.

Percutaneous transendocardial injections of therapeutic agents into the myocardium may not always be effective. We used an animal model for assessing the efficacy of the injections using linoleic acid as a testing agent. Efficacious delivery into the myocardium of a beating heart was indicated by rapidly developed local myocardial necrosis and wall motion abnormalities using echocardiography. We employed this experimental model to test our innovative technology, an acoustically active injection catheter. The Doppler ultrasound‐guided acoustically active injection catheter effectively delivers the substance to the myocardium but needs further technical improvements to minimize an unwanted systemic distribution of the agent.

Abstract Background: In Phase II trials, carfilzomib (Cfz) was associated with dyspnea (34%), hypertension (14%), renal insufficiency (24%), and peripheral edema (24%) with cardiac events reported in 7% of patients (pts). In order to better understand these toxicities, we prospectively instituted detailed cardiac evaluation in 62 Cfz treated multiple myeloma (MM) pts. Methods: Pts received Cfz between 8/11-5/14. Parameters recorded included Cfz dose, concurrent chemotherapy, hydration, blood pressure, day 1 and 2 creatinine, troponin and NT-proBNP, baseline and cycle 4 echocardiograms with EF, strain and compliance. Notable cardiac events were examined for attribution. Results: Median age was 65 years, with 60% male. 20 pts had no prior therapies, 42 were relapsed; mean 4 prior therapies (range 1-10). 20 received induction chemotherapy (Cfz, dex, thalidomide, cyclophosphamide). At relapse, Cfz was given alone in 21 pts, with added cyclophosphamide in 10, and with an IMiD in 10. Cfz dose was 20mg/m2 days 1,2 and ranged from 27-45mg/m2in subsequent treatment days. Dex was given at 20-40mg weekly in 77% pts. Hydration (250-500ml) was delivered in 89% of patients pre and 63% post treatment. Baseline hypertension (>140mmHg) was present in 20% of pts and rose ≥10mmHg in 26% on day 2, but dropped ≥10mmHg in 36%. On day 8, more pts (32%) had hypertension pre-treatment. Baseline creatinine was elevated in 25% pts. 7 of 55 (13%) pts had a creatinine rise (≥0.3 mg/dL) on day 2, 5 of whom had elevation at baseline. Thus, with normal baseline creatinine and pre-treatment hydration 2/45 (4%) pts had a creatinine rise. Troponin levels on day 1 were normal in all 16 tested patients. 2/25 pts had an elevated troponin on day 2, one with associated MI on day 4. In these 2 pts, symptomatic pre-existing ischemic heart disease and severe cardiac amyloid were confounding factors. Mean baseline NT-proBNP (n=22) was elevated at 565 pg/mL (range <50-3666; normal <77) with normal baseline values in only 41%. On day 2 (n=25), the mean was significantly higher at 1580 pg/mL. Between day 1 and 2, 72% of pts had a rise ≥50 pg/mL; mean change 1387 (range 0 to +6187). On day 2, 52% had a NT-proBNP ≥500 pg/mL, while 36% were ≥1000 pg/mL. 30 pts had baseline and post-C4 treatment echocardiograms, all with normal pre-treatment EFs. Three pts (10%) developed impaired systolic function (EF<50%), however, 2 had plausible concurrent alternative etiologies (VTE/PE, hospitalized multilobar pneumonia) while one patient (during C2) had an asymptomatic unexplained decline in EF (56 to 42%) detected pre-op while hospitalized for hip fracture. Thus, 1/30 (3%) had an EF change in absence of concurrent acute illness. A fourth patient had normal echo at C4 but was hospitalized at C10 with severe sepsis and a diminished EF was documented. All 4 pts with low EF were pre-treated (≥2 therapies plus SCT). Changes in cardiac strain often precede an EF fall. Only 14/25 pts had normal baseline cardiac strain (≥ -18%). Post treatment strain measurement significantly improved in 4 pts and worsened in 4 pts, the latter accompanied by EF decline within the normal range in 2 pts (68/60%, 65/58%). 28 pts had E/e’ ratio (LVEDP) reviewed as a marker of ventricular compliance. 46% had abnormal ratios (≥ 15), prior to Cfz treatment. 20% of patients experienced worsening parameters post treatment, however, an equal number of pts demonstrated improvement. There was no consistent correlation with change in EF. Conclusions: MM pts frequently have baseline elevated cardiac peptides (59%) and abnormal cardiac strain (15% of newly diagnosed and 36% at relapse). A frequent, and sometimes dramatic, rise in the NT-proBNP occurs immediately following Cfz based chemotherapy. Acute structural cardiac events were however uncommon (3%) in the absence of confounding illness. Overall 5/62 (8%) patients had serious cardiac events during treatment with Cfz (4 EF decline, 1 MI). These toxicities were considered probably attributable to Cfz in 3/62 (5%) patients. Prospective controlled studies with longer term follow up and bortezomib treated controls are necessary to accurately document the cardiovascular impact of Cfz and the role of proteasome inhibition. Disclosures Off Label Use: bortezomib was used in combination therapy to treat relapsed low grade lymphomas and Waldenstrom's macroglobulinemia. Fonseca:Medtronic, Otsuka, Celgene, Genzyme, BMS, Lilly, Onyx, Binding Site, Millennium, AMGEN: Consultancy, patent for the prognostication of MM based on genetic categorization of the disease. He also has sponsored research from Cylene and Onyx Other, Research Funding. Bergsagel:Novartis: Research Funding; Constellation Pharmaceutical: Research Funding; OncoEthix: Research Funding; MundiPharma: Research Funding. Reeder:Millennium, Celgene, Novartis: Research Funding. Mikhael:Onyx: Research Funding; Celgene: Research Funding; Sanofi: Research Funding; Novartis: Research Funding. Stewart:Celgene Novartis Array BioPharma BMS, research funding Millenium: Consultancy, Research Funding.

B‐mode ultrasound imaging guidance of cannulas can be compromised by noise, artifacts, and echogenicity that is not distinctive from that of surrounding anatomy. We have modified a venovenous extracorporeal membrane oxygenation cannula by embedding piezoelectric crystals into each of its 3 blood flow ports. Each vibrating crystal acoustically interacts with a Doppler imaging signal and produces an instantaneous color marker. The aim of this study was to compare identification of the extracorporeal membrane oxygenation cannula ports by B‐mode imaging versus the color Doppler marker. Unlike B‐mode imaging, the color Doppler marker identified the corresponding port even in highly challenging closed‐chest scans in anesthetized pigs. The method could improve guidance accuracy of cannulas by ultrasound scans.

Minimally invasive procedures using ultrasound are becoming prominent, especially in applications like biopsy, catheter guidance, and surgeries. In our previous work a catheter tip is fitted with a miniature omnidirectional piezoelectric crystal; the ultrasonic field from the imaging array and the piezoelectric crystal interact resulting in a symmetric Doppler shift. Previously, a frequency domain algorithm was used to identify pixels having the symmetric Doppler shift. The identified pixels can then be visualized on B-mode and color map with a unique color thus representing the location of the catheter tip. However, frequency domain algorithms are computationally expensive and require changes to data workflow. Using mean frequency estimators with symmetric Doppler shift results in the catheter tip being displayed as zero mean noise making it hard to distinguish from color Doppler noise. To overcome this limitation, complex demodulation can be used. Modulating the time domain signal with sine and cosine by the piezoelectric excitation frequency and summing them up results in cancellation of one of the symmetric Doppler components. With only one frequency component remaining mean frequency estimators can be used to identify the location of the catheter. The presented algorithm has lower computational complexity and requires minimal change in software design.

Opening a chest in an experimental echocardiographic animal study eliminates ultrasound signal attenuation by the chest wall. We developed a scanning technique that involves the use of an attenuative pad created from a mixture of urethane and titanium dioxide. The pad was interposed within transmission gel between the transducer face and cardiac surface in open‐chest scans in a porcine model. Comparative measurements of left ventricular echogenicity without and with the pad demonstrate that the pad reproducibly causes ultrasound signal attenuation that closely mimics chest attenuation in clinical transthoracic echocardiographic studies.