Predicting venous insufficiency in flaps raised on the deep inferior epigastric system using computed tomography (CT) angiography.

Computed Tomography Angiogram (CTA) has become a routine part of pre-operative assessment of vascular anatomy and design in perforator flaps. We conducted a retrospective cohort study of flap raised on the deep inferior epigastric system (DIES) at our institution in order to identify CTA signs that might predict venous congestion in these flaps. 98 consecutive patients who had 124 DIES flaps raised from 2008 to 2012 were studied. Of these 124 flaps, four (3.2%) developed venous congestion. Our results showed that a Superficial Inferior Epigastric Vein (SIEV) that is larger than the DIEV at origin is highly predictive of congestion (5.2 vs 3.5 mm, p = 0.007). The findings of an axial non-arborising superficial system (96.7% vs 0, p < 0.001), without connection to deep system perforators (38.1 vs 88.8%, p < 0.001) and a type I pedicle were also predictive (75 vs 64.2%, p = 0.22). These results show the importance of CTAs as a pre-operative study for the identification of risk factors for venous compromise, and their use should prompt a robust discussion of the risk of flap failure with patients, and contingency planning to augment venous drainage with the superficial system if required.

Effects of brain-derived neurotrophic factor on dopaminergic function and motor behavior during aging.

Brain-derived neurotrophic factor (BDNF) is critical in synaptic plasticity and in the survival and function of midbrain dopamine neurons. In this study, we assessed the effects of a partial genetic deletion of BDNF on motor function and dopamine (DA) neurotransmitter measures by comparing Bdnf(+/-) with wildtype mice (WT) at different ages. Bdnf(+/-) and WT mice had similar body weights until 12 months of age; however, at 21 months, Bdnf(+/-) mice were significantly heavier than WT mice. Horizontal and vertical motor activity was reduced for Bdnf(+/-) compared to WT mice, but was not influenced by age. Performance on an accelerating rotarod declined with age for both genotypes and was exacerbated for Bdnf(+/-) mice. Body weight did not correlate with any of the three behavioral measures studied. Dopamine neurotransmitter markers indicated no genotypic difference in striatal tyrosine hydroxylase, DA transporter (DAT) or vesicular monoamine transporter 2 (VMAT2) immunoreactivity at any age. However, DA transport via DAT (starting at 12 months) and VMAT2 (starting at 3 months) as well as KCl-stimulated DA release were reduced in Bdnf(+/-) mice and declined with age suggesting an increasingly important role for BDNF in the release and uptake of DA with the aging process. These findings suggest that a BDNF expression deficit becomes more critical to dopaminergic dynamics and related behavioral activities with increasing age.

Amphetamine-induced locomotion and gene expression are altered in BDNF heterozygous mice.

Administration of amphetamine overstimulates medium spiny neurons (MSNs) by releasing dopamine and glutamate from afferents in the striatum. However, these afferents also release brain-derived neurotrophic factor (BDNF) that protects striatal MSNs from overstimulation. Intriguingly, all three neurochemicals increase opioid gene expression in MSNs. In contrast, striatal opioid expression is less in naive BDNF heterozygous (BDNF(+/-)) vs. wild-type (WT) mice. This study was designed to determine whether partial genetic depletion of BDNF influences the behavioral and molecular response to an acute amphetamine injection. An acute injection of amphetamine [5 mg/kg, intraperitoneal (i.p.)] or saline was administered to WT and BDNF(+/-) mice. WT and BDNF(+/-) mice exhibited similar locomotor activity during habituation, whereas BDNF(+/-) mice exhibited more prolonged locomotor activation during the third hour after injection of amphetamine. Three hours after amphetamine injection, there was an increase of preprodynorphin mRNA in the caudate putamen and nucleus accumbens (Acb) and dopamine D(3) receptor mRNA levels were increased in the Acb of BDNF(+/-) and WT mice. Striatal/cortical trkB and BDNF, and mesencephalic tyrosine hydroxylase mRNA levels were only increased in WT mice. These results indicate that BDNF modifies the locomotor responses of mice to acute amphetamine and differentially regulates amphetamine-induced gene expression.

Contribution of the mitochondrial permeability transition to lethal injury after exposure of hepatocytes to t-butylhydroperoxide.

We have developed a novel method for monitoring the mitochondrial permeability transition in single intact hepatocytes during injury with t-butylhydroperoxide (t-BuOOH). Cultured hepatocytes were loaded with the fluorescence probes, calcein and tetramethylrhodamine methyl ester (TMRM). Depending on loading conditions, calcein labelled the cytosolic space exclusively and did not enter mitochondria or it stained both cytosol and mitochondria. TMRM labelled mitochondria as an indicator of mitochondrial polarization. Fluorescence of two probes was imaged simultaneously using laser-scanning confocal microscopy. During normal incubations, TMRM labelled mitochondria indefinitely (longer than 63 min), and calcein did not redistribute between cytosol and mitochondria. These findings indicate that the mitochondrial permeability transition pore ('megachannel') remained closed continuously. After addition of 100 microM t-BuOOH, mitochondria filled quickly with calcein, indicating the onset of mitochondrial permeability transition. This event was accompanied by mitochondrial depolarization, as shown by loss of TMRM. Subsequently, the concentration of ATP declined and cells lost viability. Trifluoperazine, a phospholipase inhibitor that inhibits the permeability transition in isolated mitochondria, prevented calcein redistribution into mitochondria, mitochondrial depolarization, ATP depletion and cell death. Carbonyl cyanide m-chlorophenylhydrazone (CCCP), a mitochondrial uncoupler, also rapidly depolarized mitochondria of intact hepatocytes but did not alone induce a permeability transition. Trifluoperazine did not prevent ATP depletion and cell death after the addition of CCCP. In conclusion, the permeability transition pore does not 'flicker' open during normal incubation of hepatocytes but remains continuously closed. Moreover, mitochondrial depolarization per se does not cause the permeability transition in intact cells. During oxidative stress, however, a permeability transition occurs quickly which leads to mitochondrial depolarization and cell death.

ATP depletion rather than mitochondrial depolarization mediates hepatocyte killing after metabolic inhibition.

The importance of ATP depletion and mitochondrial depolarization in the toxicity of cyanide, oligomycin, and carbonyl cyanide m-cholorophenylhydrazone (CCCP), an uncoupler, was evaluated in rat hepatocytes. Oligomycin, an inhibitor of the reversible mitochondrial ATP synthase (F1F0-adenosinetriphosphatase), caused dose-dependent cell killing with 0.1 microgram/ml being the minimum concentration causing the maximum cell killing. Oligomycin also caused rapid ATP depletion without causing mitochondrial depolarization. Fructose (20 mM), a potent glycolytic substrate in liver, protected completely against oligomycin toxicity. CCCP (5 microM) also caused rapid killing of hepatocytes. Fructose retarded cell death caused by CCCP but failed to prevent lethal cell injury. Although oligomycin (1.0 microgram/ml) was lethally toxic by itself, in the presence of fructose it protected completely against CCCP-induced cell killing. Cyanide (2.5 mM), an inhibitor of mitochondrial respiration, caused rapid cell killing that was reversed by fructose. CCCP completely blocked fructose protection against cyanide, causing mitochondrial depolarization and rapid ATP depletion. In the presence of fructose and cyanide, oligomycin protected cells against CCCP-induced ATP depletion and cell death but did not prevent mitochondrial depolarization. In every instance, cell killing was associated with ATP depletion, whereas protection against lethal cell injury was associated with preservation of ATP. In conclusion, protection by fructose against toxicity of cyanide, oligomycin, and CCCP was mediated by glycolytic ATP formation rather than by preservation of the mitochondrial membrane potential. These findings support the hypothesis that inhibition of cellular ATP formation is a crucial event in the progression of irreversible cell injury.

The effects of curvature on fluid flow fields in pulmonary artery models: flow visualization studies.

In vitro pulsatile flow visualization studies were conducted to assess the effects of varying radii of curvature of the right ventricular outflow tract (RVOT) and main pulmonary artery (MPA) on the flow fields in the main, right, and left pulmonary arteries of a one month lamb pulmonary artery model. Three glass flow-through models were studied; one with no curvature, one with the correct anatomic curvature, and one with an overaccentuated curvature on the RVOT and MPA. All other geometric parameters were held constant. Pulsatile flow visualization studies were conducted at nine flow conditions; heart rates of 70, 100, and 140 bpm, and cardiac outputs of 1.5, 2.5 and 3.5 l/min with corresponding mean pulmonary pressures of 10, 20, and 30 mmHg. Changes were observed in the pulmonary flow fields as the curvature of the outflow tract, heart rate and mean pulmonary pressure were varied. An increase in vessel curvature led to an increase in the overall radial nature of the flow field as well as flow separation regions which formed faster, originated further downstream, and occupied more of the vessel area. At higher heart rates, the maximum size of the separation regions decreased, while flow separation regions appeared earlier in the cardiac cycle and grew more quickly. Heart rate also affected the initiation of flow reversal; flow reversal occurred later in the cardiac cycle at lower heart rates. Both heart rate and mean pulmonary pressure influenced the stability of the pulmonary flow field and the appearance of coherent structures. In addition, an increase in mean pulmonary pressure increased the magnitude of reverse flow.(ABSTRACT TRUNCATED AT 250 WORDS)