Warm Circumpolar Deep Water transport toward Antarctica driven by local dense water export in canyons.

Poleward transport of warm Circumpolar Deep Water (CDW) has been linked to melting of Antarctic ice shelves. However, even the steady-state spatial distribution and mechanisms of CDW transport remain poorly understood. Using a global, eddying ocean model, we explore the relationship between the cross-slope transports of CDW and descending Dense Shelf Water (DSW). We find large spatial variability in CDW heat and volume transport around Antarctica, with substantially enhanced flow where DSW descends in canyons. The CDW and DSW transports are highly spatially correlated within ~20 km and temporally correlated on subdaily time scales. Focusing on the Ross Sea, we show that the relationship is driven by pulses of overflowing DSW lowering sea surface height, leading to net onshore CDW transport. The majority of simulated onshore CDW transport is concentrated in cold-water regions, rather than warm-water regions, with potential implications for ice-ocean interactions and global sea level rise.

Southern Hemisphere westerlies as a driver of the early deglacial atmospheric CO2 rise.

The early part of the last deglaciation is characterised by a ~40 ppm atmospheric CO2 rise occurring in two abrupt phases. The underlying mechanisms driving these increases remain a subject of intense debate. Here, we successfully reproduce changes in CO2, δ13C and Δ14C as recorded by paleo-records during Heinrich stadial 1 (HS1). We show that HS1 CO2 increase can be explained by enhanced Southern Ocean upwelling of carbon-rich Pacific deep and intermediate waters, resulting from intensified Southern Ocean convection and Southern Hemisphere (SH) westerlies. While enhanced Antarctic Bottom Water formation leads to a millennial CO2 outgassing, intensified SH westerlies induce a multi-decadal atmospheric CO2 rise. A strengthening of SH westerlies in a global eddy-permitting ocean model further supports a multi-decadal CO2 outgassing from the Southern Ocean. Our results highlight the crucial role of SH westerlies in the global climate and carbon cycle system with important implications for future climate projections.

Flow dynamics of a novel counterpulsation device characterized by CFD and PIV modeling.

BACKGROUND: Historically, single port valveless pneumatic blood pumps have had a high incidence of thrombus formation due to areas of blood stagnation and hemolysis due to areas of high shear stress. METHODS: To ensure minimal hemolysis and favorable blood washing characteristics, particle image velocimetry (PIV) and computational fluid dynamics (CFD) were used to evaluate the design of a new single port, valveless counterpulsation device (Symphony). The Symphony design was tested in 6-h acute (n=8), 5-day (n=8) and 30-day (n=2) chronic experiments in a calf model (Jersey, 76 kg). Venous blood samples were collected during acute (hourly) and chronic (weekly) time courses to analyze for temporal changes in biochemical markers and quantify plasma free hemoglobin. At the end of the study, animals were euthanized and the Symphony and end-organs (brain, liver, kidney, lungs, heart, and spleen) were examined for thrombus formations. RESULTS: Both the PIV and the CFD showed the development of a strong moving vortex during filling phase and that blood exited the Symphony uniformly from all areas during ejection phase. The laminar shear stresses estimated by CFD remained well below the hemolysis threshold of 400 Pa inside the Symphony throughout filling and ejection phases. No areas of persistent blood stagnation or flow separation were observed. The maximum plasma free hemoglobin (<10mg/dl), average platelet count (pre-implant = 473 ± 56 K/µl and post-implant = 331 ± 62 K/µl), and average hematocrit (pre-implant = 31 ± 2% and post-implant = 29 ± 2%) were normal at all measured time-points for each test animal in acute and chronic experiments. There were no changes in measures of hepatic function (ALP, ALT) or renal function (creatinine) from pre-Symphony implantation values. The necropsy examination showed no signs of thrombus formation in the Symphony or end organs. CONCLUSIONS: These data suggest that the designed Symphony has good washing characteristics without persistent areas of blood stagnation sites during the entire pump cycle, and has a low risk of hemolysis and thrombus formations.

Hemodynamic and left ventricular pressure-volume responses to counterpulsation in mock circulation and acute large animal models.

Alternative therapies for treating heart failure patients are being explored to provide effective options for patients with progressive heart failure. Cardiac assist devices that promote myocardial recovery may be a potential solution. Ventricular assist devices (VAD) have demonstrated long-term efficacy and intraaortic balloon pumps (IABP) have shown short-term successes. In this paper, testing of a hybrid counterpulsation device (CPD) that couples the attributes of device longevity (VAD) with less invasive surgery (IABP) is presented. Hemodynamic and ventricular pressure-volume responses to a 40 ml CPD and 40 ml IABP were evaluated in vitro in an adult mock circulation and in vivo in a large animal heart failure model. The CPD is a flexing diaphragm ventricle with a controlled stroke volume up to 85 cc through a single, valveless cannula. In this study, the CPD was cannulated to the brachiocephalic artery to provide 40 ml of counterpulsation support. The CPD effectively provided diastolic augmentation increasing coronary flow and afterload reduction. These results were comparable to IABP. These preliminary studies suggest that CPD may be an effective therapy for treating patients with early stage heart failure.

Syngeneic central nervous system transplantation of genetically transduced mature, adult astrocytes.

Advances in the development of highly infectious, replication-deficient recombinant retroviruses provide an efficient means of stable transfer of gene expression. Coupled with ex vivo transduction, surrogate cell populations can be readily implanted into the brain, thus serving as vehicles for delivering selected gene products into the central nervous system (CNS). Here we report that rat astrocytes can be routinely and safely isolated from brain tissue of a living donor by use of short-term gelatin sponge implants. The mature, nontransformed astrocytes were easily expanded, maintained in long-term tissue cultures and were efficiently transduced with an amphotropic retrovirus harboring a heterologous, fused transgene. In vitro retroviral infection rendered the nontransformed cells essentially 100% viable after exposure. The level of efficiency of infection (30-50% effective genome integration of provirus and expression of transgene in target cell populations) and minimal cell toxicity obviated the need to harvest large numbers of target cells. Cultured transduced astrocytes were resilient and exhibited select peptide expression for up to 1 year. Subsequently, transduced astrocytes were used in a series of experiments in which cells were transplanted intracerebrally in syngeneic animals. Post-implantation, astrocytes seeded locally and either insinuated into the surrounding parenchyma in situ or exhibited a variable degree of migration, depending on the anatomic source of astrocytes and the targeted brain implantation site. Transduced astrocytes remained viable in excess of 8 months post-transplantation and exhibited sustained transgenic peptide expression of green fluorescent protein/neomycin phosphotransferase in vivo. The sequential isolation and culture of nontransformed, mature, adult astrocytes and recombinant retrovirus-mediated transduction in vitro followed by brain reimplantation represents a safe and effective means for transferring genetic expression to the CNS. This study lays the foundation for exploring the utility of using a human autologous transplantation system as a potential gene delivery approach to treat neurological disorders. Prepared and utilized in this manner, autologous astrocytes may serve as a vehicle to deliver gene therapy to the CNS.

Generation of cellular immunity to lymphocytic choriomeningitis virus is independent of CD1d1 expression.

CD1 molecules are cell surface glycoproteins, structurally similar to major histocompatibility complex (MHC) class I molecules. The murine CD1d1 molecule has been shown to be essential for the positive selection of a unique subpopulation of T cells [the natural killer (NK) T cells], as CD1d1-deficient mice lack NK T cells. These cells have recently been suggested to play an important role in the induction of innate immunity (i.e. NK cells) and the regulation of immune homeostasis. As such, it was asked whether NK T cells were necessary for the generation of cellular immunity to an acute virus infection. In these studies, the Armstrong strain of lymphocytic choriomeningitis virus (LCMV), a classic inducer of NK cells, and its pathogenic variant clone 13 were used. When NK-cell activity was assessed on day 3 post-LCMV infection, surprisingly, it was found that CD1d1-deficient mice could generate NK-cell activity at wild-type levels. Likewise, LCMV-specific cytotoxic T-lymphocyte (CTL) activity in CD1d1-deficient mice was indistinguishable from that generated in wild-type mice. Additionally, viral titres in the spleen (LCMV Armstrong) and blood (LCMV clone 13) of infected CD1d1-deficient mice were at comparable levels to those found in wild-type mice, as were virus infection-induced increases in cell surface H-2Kb in the spleen. Therefore, these results suggest that the LCMV-induced generation of NK-cell and virus-specific CTL activity, as well as viral clearance, are independent of CD1d1 expression.

Successful VATS ligation of a large anomalous branch producing IMA steal syndrome after MIDCAB.

The occurrence of a flow "steal phenomenon" from a large branch of the internal mammary artery (IMA) is rare and its pathogenesis is still controversial. We describe a case of a patent large anomalous left IMA branch which produced recurrent angina 3 years post MIDCAB. Transcatheter coil obliteration of the vessel produced symptomatic relief. However, six months later, chest pains recurred in association with unwinding of the coil and recannulization of the collateral vessel. Successful ligation of the culprit branch using a VATS approach is presented.

Lethal domestic violence in eastern North Carolina.

Strategies for preventing domestic violence can be tailored to a particular geographic or socioeconomic area if the patterns of domestic violence in the area are known. National statistics, although widely available, may not be applicable to a specific region. We reviewed homicide deaths in Eastern North Carolina between 1978 and 1999 to identify patterns in this rural area. Approximately 20% of the homicide deaths in eastern North Carolina are caused by intimate partners. Women accounted for 53% of the victims in 1976, similar to national figures but not rising to 72% as seen nationally in 1998. Latinos are an increasing presence in the area, but had only one recorded episode of lethal violence against an intimate partner. Gunshots accounted for most of the deaths (59% in men, 72% in women). Knowledge of such patterns can assist in selecting prevention strategies for this particular area. Over the last 25 years increasing attention has been devoted to domestic violence (DV), initially defined as abuse committed against a spouse, former spouse, fiancée, boy- or girlfriend, or cohabitant. As time has passed, the definition has been broadened to include other family members--elders, children, and siblings. The Centers for Disease Control and Prevention (CDC) now uses the term "intimate partner violence" for intentional emotional or physical abuse inflicted by a spouse, ex-spouse, a present or former boy- or girlfriend, or date. For the purposes of this paper, we consider DV interchangeable with intimate partner violence. There has been a national concern that abusive events are under-reported. The National Crime Victimization Survey, an anonymous household survey, indicated nearly 1 million incidents of non-lethal intimate partner violence per year between 1992 and 1996. The number decreased from 1.1 million in 1993 to 840,000 in 1996. Attempts to validate such data for a given geographic area often require subjects to violate anonymity--this may account for lower reports of violence. A recent national report from the Justice Department found a decline in both lethal and non-lethal DV. The number of men murdered by wives or girlfriends "plunged 60% from 1976 through 1998". FBI data on homicides showed that "intimate partners committed fewer murders each year during 1996, 1997, and 1998 than in any other year since 1976". Nationally, intimate partners caused 3000 deaths in 1976, 1590 (53%) in women; in 1998, they caused 1830 deaths, 1320 (73%) in women. But fatal cases of DV are only the tip of the iceberg, and may pertain only to a particular geographic area. We undertook the present study to assess the prevalence of lethal domestic violence in the 29 counties of eastern North Carolina (ENC) that make up the catchment area for the University Health Systems of Eastern Carolina (UHSEC). UHSEC includes the Brody School of Medicine at East Carolina University (BSM-ECU; previously known as East Carolina University-SOM) and Pitt County Memorial Hospital.

Investigation of capillary electrochromatography with brush-type chiral stationary phases.

Fused-silica capillaries (100 microm ID) were packed with the (3R, 4S)-Whelk-O chiral stationary phase (CSP) bonded on 3.0 microm silica particles. The enantiomers of 41 neutral analytes containing stereogenic centers, axes or planes were examined by packed capillary electrochromatography. More than 30 of these were cleanly resolved, owing to the selectivities and efficiencies afforded by this CSP. High reproducibility with no indication of diminished performance was observed using the same capillary for hundreds of runs (including intermediate change of the buffer system) over a period of several weeks. Acetate, 2-(N-morpholino)ethanesulfonic acid, or phosphate buffers, each modified with either acetonitrile or methanol, were used as mobile phases. The influence of buffer concentration, modifier amount, temperature, applied voltage, and pH on performance of the brush-type CSP was investigated.

A new technique to create an artificial stenosis in the native LAD using a hemoclip.

The study of hemodynamics associated with coronary atherosclerotic stenosis has been limited due to the lack of a safe, accurate, and reliable technique to create an artificial stenosis in an animal model. Existing techniques have often resulted in myocardial infarction (MI) or severe injury to the vessel and have been difficult to accurately quantify and reproduce. We developed a new technique to create an artificial stenosis in the native left anterior descending (LAD) coronary artery using a hemoclip in an experimental off-pump coronary artery bypass graft (CABG) animal model that overcomes these limitations. The native LAD was dissected at its proximal end and a hemoclip was applied to create varying degrees of artificial stenosis in 30 mongrel dogs during experimental off-pump CABG procedures. The precise application of the hemoclip was predetermined using a mathematical formula to calculate the reduced circumference required to create a specific stenosis. Using these calculations, artificial stenoses of 25%, 50%, 75%, and 90% were created in the LAD. Postoperative angiography demonstrated only 5-10% error between the true (angiography) and estimated (hemoclip technique) stenosis values. In all cases, the vessel remained intact without any apparent external trauma, and no indications of MI were present during electrocardiograph (ECG) monitoring. The creation of an artificial coronary stenosis using the hemoclip technique was safe, reliable, easy, and accurate.

Spectral analysis of graft flow for anastomotic error detection in off-pump CABG.

OBJECTIVE: Flow probes have been introduced as a non-invasive means of anastomotic quality assessment in off-pump coronary artery bypass graft (CABG). Flow waveform morphology cannot reliably be assessed visually unless severe anastomotic stenosis is present ( > 90%). We applied spectral analysis techniques to determine whether the frequency content of graft flow can improve the surgeon's ability to detect anastomotic errors. METHODS: Forty-six mammary to left anterior descending artery (LAD) anastomoses were created in mongrel dogs during off-pump CABG surgery. Graft flow was measured using transit-time flow probes with the LAD closed, and the mammary graft patent and with varying degrees of stenosis. The degree of anastomotic stenosis was created by an artificial stitch and verified by random postoperative angiography. Spectral analysis of the graft flow waveforms was performed. Differences in the magnitude and phase components of the graft flow for the first five harmonics were determined for the varying anastomosis test conditions. Differences were determined using analysis of variance and least square means techniques. RESULTS: The magnitude of the fundamental (zeroth) harmonic was statistically different in the internal mammary artery (IMA) with 0-25% stenosis compared to IMA with 50-75% stenosis (P < 0.01 ). Further, the magnitude of the first, second, and fourth harmonics were statistically different in IMA with 0-25% compared to IMA with 75% (P < 0.01). The phase of the first harmonic was statistically different in IMA with 25% stenosis than IMA with 50% stenosis (P < 0.01 ). No differences in interaction between the LAD and IMA for all ranges of stenosis were detected (P > 0.50). CONCLUSION: Spectral analysis of graft flow waveforms may be beneficial in detecting lesser degrees of anastomotic stenosis (i.e. < 90%) compared to traditional visual assessment of mean graft flow and/or graft flow waveform morphology.

Neural network pattern recognition analysis of graft flow characteristics improves intra-operative anastomotic error detection in minimally invasive CABG.

OBJECTIVE: The intra-operative assessment of the quality of anastomosis in minimally invasive coronary artery bypass surgery (CABG) is critical. Recent investigations demonstrated that flow probes used intra-operatively to assess anastomotic errors may give the surgeon a false sense of confidence as only severely stenotic anastomoses (>90%) could be reliably detected. We developed a neural network system using graft flow data and assessed its potential to improve anastomotic error detection. METHODS: Mammary to LAD grafts (n = 46) were constructed in mongrel dogs off-pump. Continuous beat-to-beat graft flow was recorded using transit-time flow probes. Various degrees of anastomotic stenoses (0-100%) were created by an additional suture. The degree of anastomotic stenosis was confirmed by postoperative angiography. A learning vector quantization neural network was created using heart rate, mean aortic pressure, mean systolic, maximum systolic, minimum systolic, mean diastolic, maximum diastolic, minimum diastolic, and mean graft flows. In addition, a spectral analysis of the flow waveforms was performed and the magnitude and phase of the first five harmonics were used to further develop the neural network. RESULTS: The neural network pattern recognition system was 94% accurate in detecting any stenosis >50%. To validate the model, a testing set was used with 20% of the data values, and the accuracy remained at 100% above chance alone. CONCLUSION: Pattern recognition of transit-time flow probe tracings using neural network systems can detect anastomotic errors significantly better than the surgeon's visual assessment, thereby improving the clinical outcome of minimally invasive CABG.

Genomics: the race is on.

'ellipsisgenomics has started a race in the pharmaceutical and biotechnology industryellipsis'

A review of transit-time flow measurement for assessing graft patency.

Anastomotic quality is a critical issue in minimally invasive coronary artery bypass surgery, particularly "off-pump". It is important to detect a "poor" anastomosis during the procedure so future re-operation can be avoided. Methods such as intraoperative angiography, thermal angiography, probing of the anastomosis, and graft flow measurement have been used intraoperatively to help identify anastomotic errors. With the evolution of stabilizers, graft patency rates for off-pump cases have improved, but many believe they are still not as high as those of the conventional procedure. For off-pump surgery to be accepted and practiced universally, patency rates must be equivalent to those of "on-pump" cases. Transit-time flow measurement has become an increasingly popular non-invasive method for assessing anastomotic quality. However, it is difficult to establish whether an anastomosis is patent based on mean graft flow alone. Spectral analysis of graft flow waveforms reveal characteristic patterns that identify intermediate ranges of stenosis between fully patent and totally occluded. Together, these two components of graft flow have been used in the construction of a neural network to help identify "faulty" anastomoses. Transit-time flow measurement is a non-invasive tool that can be beneficial in identifying fully patent or nearly occluded grafts, and may also help in distinguishing intermediate stenoses.

From genome to drug--optimising the drug discovery process.

Current drug discovery and development practices are technologically sophisticated and highly efficient. At the same time the failure rate of compounds in both preclinical and clinical development is high. These failures can be attributed to many factors. Two predominant causes of failure are lack of efficacy and toxicity. Often lack of efficacy is only determined late in the clinical trial process and can be difficult if not impossible to explain, as well as being expensive. Toxicity accounts for many failures during preclinical development, which are less costly, but it also occurs in the clinic. Often the underlying cause of clinical toxicity is never identified. Studies of the structure and activity of the human and other genomes has over the last decade lead to a revolution in biological and medical research. Disease associated genes can now be identified through the application of human genetics, whole genomes have been sequenced and tools have been developed that allow the complete characterization of an organism's gene expression profile in a single experiment. These tools are now being applied to pharmaceutical research and development with the aim to increase the efficiency of the process and the quality of the product.

Can visual assessment of flow waveform morphology detect anastomotic error in off-pump coronary artery bypass grafting?

OBJECTIVE: Flow probes are being used for intraoperative assessment of anastomotic quality during off-pump coronary artery bypass grafting (CABG). We conducted a survey with the cooperation of 19 international surgeons to assess the ability of surgeons to detect anastomotic errors by evaluating mean flow and flow waveform morphology. MATERIAL AND METHODS: Mongrel dogs underwent mammary to left anterior descending (LAD) grafting. Mean graft flow and flow wave morphology for varying degrees of anastomotic stenoses were recorded using transit-time flow probes. A questionnaire consisting of ten different recorded flow tracings and the corresponding mean flows were given to 20 surgeons from around the world. The surgeons were asked to determine the degree of stenosis and whether they would re-do the anastomosis based upon the mean flow and the flow tracings. RESULTS: All of the 19 surgeons that responded were able to clearly identify a highly stenotic graft (>90% stenosis). However, 24% would re-do a fully patent anastomosis, 58% accepted an anastomosis with moderate stenosis, and 72% accepted anastomoses with severe stenosis. CONCLUSIONS: Evaluation of flow tracing morphology and/or mean flows can be used to reliably detect nearly occluded anastomoses (>90% stenosis). However, surgeons should be cautious in assessing anastomoses with lesser degrees of stenosis, as they may be more difficult to reliably interpret.

Structure-activity relationships of the Kvbeta1 inactivation domain and its putative receptor probed using peptide analogs of voltage-gated potassium channel alpha- and beta-subunits.

Certain beta-subunits exert profound effects on the kinetics of voltage-gated (Kv) potassium channel inactivation through an interaction between the amino-terminal "inactivation domain" of the beta-subunit and a "receptor" located at or near the cytoplasmic mouth of the channel pore. Here we used a bacterial random peptide library to examine the structural requirements for this interaction. To identify peptides that bind Kv1.1 we screened the library against a synthetic peptide corresponding to the predicted S4-S5 cytoplasmic loop of the Kv1.1 alpha-subunit (residues 313-328). Among the highest affinity interactors were peptides with significant homology to the amino terminus of Kvbeta1. We performed a second screen using a peptide from the amino terminus of Kvbeta1 (residues 2-31) as "bait" and identified peptide sequences with significant homology to the S4-S5 loop of Kv1.1. A series of synthetic peptides containing mutations of the wild-type Kvbeta1 and Kv1.1 sequences were examined for their ability to inhibit Kvbeta1/Kv1.1 binding. Amino acids Arg20 and Leu21 in Kvbeta1 and residues Arg324 and Leu328 in Kv1.1 were found to be important for the interaction. Taken together, these data provide support for the contention that the S4-S5 loop of the Kv1.1 alpha subunit is the likely acceptor for the Kvbeta1 inactivation domain and provide information about residues that may underlie the protein-protein interactions responsible for beta-subunit mediated Kv channel inactivation.

Role of graft flow measurement technique in anastomotic quality assessment in minimally invasive CABG.

BACKGROUND: Anastomotic quality is currently the critical issue in minimally invasive coronary surgery. Although little is known about its effectiveness, surgeons routinely assess grafts intraoperatively using flow probes. This study was designed to determine whether mean flow and the pattern of flow tracing in internal mammary artery grafts obtained with a transit-time flow probe are reliable indicators of anastomotic quality. METHODS: Mongrel dogs (n = 14, 30 to 35 kg) underwent off-pump left, right, or left and right internal mammary artery to left anterior descending artery anastomosis (23 grafts). Moderate to severe degrees of stenosis were created at the anastomosis by an additional suture. Internal mammary artery graft flow was measured before and after the stenosis was created with the left anterior descending artery occluded. Angiography was performed at random postoperatively to validate the degree of stenosis. Mean flow and flow tracing morphology were compared under various degrees of stenosis. RESULTS: There were no significant differences in mean graft flow or the morphology of the flow tracing between patent (<15%), mild (<25%), moderate (<50%), and moderately severe (<75%) stenosis. However, mean graft flow decreased (p < 0.05) with severe stenosis (>75%). CONCLUSIONS: Although differences in mean graft flow and graft flow morphology were detectable in anastomoses with severe stenosis (>75%), they were indistinguishable in anastomoses with mild (<25%) to moderately severe (<75%) stenosis. Flow measurement techniques are valuable tools intraoperatively, but surgeons should exercise caution in their interpretation.