Ultrasound: a reproducible method to measure conduit vein compliance.

Classical venous occlusion plethysmography (VOP) of the leg, often used to assess venous compliance, measures properties of the whole calf, including volume changes at the arterial side and the interstitial fluid accumulation that occurs as a result of the enhanced capillary pressure during venous occlusion. We present an ultrasound technique to measure the compliance of one major conduit vein in the leg. Ultrasound measurements of the popliteal vein were compared with classical VOP measurements, which were performed simultaneously in one subject. Six healthy individuals were measured on three occasions to assess short- and long-term reproducibility of the measurements. Six motor complete spinal cord-injured (SCI) individuals were included to compare venous compliance in subjects with known pathological changes of the venous system with controls. The ultrasound and VOP measurements of venous compliance correlated significantly (r(2) = 0.39, P = 0.001). Ultrasound provides reproducible measurements with short- and long-term coefficients of variation ranging from 10 to 15% for popliteal vein compliance and from 2 to 9% for absolute diameters at the different venous pressure steps. In addition, by using ultrasound, we were able to detect an 80% reduction in the compliance of the popliteal vein in SCI individuals compared with controls (P < 0.01). In conclusion, ultrasound is a suitable and reproducible method to measure conduit vein compliance and provides the possibility to specifically assess compliance of one vein instead of the whole calf.     

Inhomogeneity during deflation of excised canine lungs. II. Alveolar volumes

We have previously demonstrated appreciable inhomogeneity of alveolar pressures measured by a capsule technique in excised canine lobes deflated at submaximal flows (J. Appl. Physiol. 65: 1757-1765, 1988). We further analyzed the results of these experiments by estimating alveolar volumes (VA) and regional flows from regional transpulmonary pressures, assuming that regional pressure-volume relationships were homogeneous. Deflation at submaximal flows of lungs suspended in air caused significant flow-dependent inhomogeneity of VA that increased as lung volume decreased. Immersion of lungs in stable foams that simulated the gradient of pleural pressure modified the pattern of emptying, but not always to a gravity-dependent sequence. Limitation of regional expiratory flow was often asynchronous during both air suspension and foam immersion. There was no evidence of a common regional flow-volume curve. Submaximal deflation is a complex heterogeneous process, with the interregional pattern of emptying determined by the interaction of factors that are both intrinsic and extrinsic to the lungs.     

Inhomogeneity during deflation of excised canine lungs. I. Alveolar pressures

Factors both intrinsic and extrinsic to the lung may cause inhomogeneity of alveolar pressures during deflation. Wilson et al. (J. Appl. Physiol. 59: 1924-1928, 1985) predicted that any such inhomogeneity would be limited by interdependence of regional expiratory flows. To test this hypothesis and to explore how the pleural pressure gradient might affect inhomogeneity of alveolar pressures, we deflated at submaximal flows excised canine lobes that first were suspended in air and then were immersed in foams that simulated the vertical gradient of pleural pressure. Interregional inhomogeneity of regional transpulmonary pressures was measured with use of an alveolar capsule technique. Flow-dependent inhomogeneity of alveolar pressures was present, with differences in alveolar pressure quickly relaxing to a constant limiting value at each flow. Foam immersion increased inhomogeneity at a given flow. We conclude that factors intrinsic to the lung cause significant inhomogeneity of alveolar pressures at submaximal expiratory flows and that this inhomogeneity is enhanced by the extrinsic gradient of pleural pressure. These observations are consistent with the interdependence of flow proposed by Wilson et al.     

Inhomogeneity during deflation of excised canine lungs. III. Single-breath O2 tests

Both interregional and intraregional mechanisms may cause changes in N2 concentration of expired gas during the phases of the single-breath O2 test (SBO2) that follow dead-space washout. To evaluate the possible importance of each mechanism, we performed the SBO2 in excised canine lungs that were first suspended in air and then immersed in stable foams that simulated the vertical gradient of pleural pressure. The lungs were deflated at constant submaximal flows. The slope of phase III diminished with increasing expiratory flow and increased with foam immersion. The onset of phase IV depended on flow, and a terminal decrease in N2 concentration (phase V) was often observed. Simultaneously measured estimates of regional flows and volumes (J. Appl. Physiol. 65: 1764-1774, 1988) were used to further interpret these results. The onset of phase IV at flows greater than quasi-static signified the onset of flow limitation of dependent regions. The onset of phase V corresponded to flow limitation of nondependent regions.     

Placental compliance during fetal extracorporeal circulation.

The fetus requires large amounts of volume when weaning from cardiac bypass. This suggests that placental vasculature can act as a large capacitor in the fetal circulation. To assess placental compliance of fetal lambs, seven isolated in situ lamb placentas were placed on extracorporeal circulation. Umbilical artery blood flow was varied from 0 to 350 ml. min(-1). kg fetal wt(-1). Because the extracorporeal circuit is a closed system, volume changes in the placenta induced by umbilical artery pressure changes were measured from reciprocal volume changes in the reservoir. There was a wide range of change in absolute volume of blood within the fetal placental compartment (216.4 +/- 29.3 ml). Placental compliance was linear over the entire range of pressure changes exerted on the placental vasculature (r(2) = 0.83, P = 0.0001). This indicates that the placenta is a unique and sensitive capacitor in the fetal circulation. This information is important clinically because it establishes that aggressive resuscitation of the fetus using volume may be necessary when weaning the fetus from cardiac bypass.     

Biological indicators provide short term soil health assessment during sodic soil reclamation

Sodic soil remediation is an expensive, lengthy process during which producers need tools to demonstrate that sodium (Na⁺) remediation practices are improving soil health. The objective of this study was to determine if soil biological indicators can provide a short term assessment of the effectiveness of chemical management strategies used to remediate northern Great Plains sodium affected soils. This randomized complete block split-plot research experiment was conducted in a grassland which was converted to annual row crops. The soil at the site was an Exline (fine, smectitic, frigid Leptic Natrudolls). The experiment contained two drainage treatments (tile drained and no-drainage) and four chemical amendments (4.5 Mg ha⁻¹ of gypsum, 9.1 Mg ha⁻¹ of gypsum, 9.1 Mg ha⁻¹ spent sugar beet lime, and a no amendment control). Base-line soil samples for biological assessment were collected in the fall of 2012 after tile drainage was installed. The sodium adsorption ratio (SAR) ranged from 0.4 to 16.7 with a range of electrical conductivity (EC) of 0.4–0.8 dS m⁻¹. Gypsum and lime amendments were applied in 2013. Soil samples were collected for assessing soil health before and after application of amendments and throughout the growing season. This study utilizes a novel application of successional vector trajectories to compare shifts in measured soil health parameters associated with land use change and remediation of sodicity. Soil samples were analyzed for percent total soil carbon (C), nitrifier and denitrifier gene copies, soil enzyme assays (nitrate reductase, ammonia monooxegenase, urease, β glucosidase, alkaline phosphatase, arylsulfatase and fluorescein diacetate hydrolysis), EC, pH, SAR, and soil texture. Gene copies and enzyme activities were successfully used to differentiate between land uses and amendment applications. Ammonia oxidizing bacterial gene copies were higher where cropland was amended with gypsum. Successional vectors verified a significant shift in soil health due to land use change and amendment application. Gypsum applications reduced SAR, but increased soil EC. This work demonstrates that soil enzyme activities and gene copy numbers can be used to detect improvements in soil health.     

A short protocol for micro-purification of nuclear proteins from whole animal tissue.

Although a number of small-scale procedures have been described for the preparation of crude nuclear extracts from established cell lines, none were provided for the preparation of similar extracts from small amounts of animal tissue. In addition, no small-scale procedures contain enrichment steps that render the detection of low-abundant DNA-binding proteins easier. Here we describe a simple, efficient procedure for the rapid preparation of high-quality nuclear extracts from either whole animal tissue or established cell lines. It is based on a rapid isolation of the nuclei followed by a KCl extraction and a further micro-enrichment of the DNA binding proteins on heparin Sepharose CL-6B. Extracts prepared in such a way are suitable for the analysis of specific DNA/protein interactions by the use of gel shift assays or by DNaseI and dimethylsulfate footprinting techniques. Most importantly, the entire process can be fulfilled at minimal cost within a day on as little as one gram of fresh tissue, which renders this procedure extremely attractive for the analysis of DNA binding proteins involved in the control of gene expression.     

Ultrasound monitoring of hepatic metastases during chemotherapy.

Grey-scale ultrasonography has been found to be the most sensitive method of detecting metastatic disease of the liver. In two cases the results of chemotherapy were monitored by ultrasound; the response to treatment could be distinguished from non-response and ultrasonography gave useful information when chemotherapy made radioisotope examination unreliable.     

Mechanism of lobar alveolar pressure decline during forced deflation in canine regional emphysema

Mink, S. N. Mechanism of lobar alveolar pressuredecline during forced deflation in canine regional emphysema.J. Appl. Physiol. 82(2): 632-643, 1997.A canine model of unilobar papain-induced emphysema was used toexamine the extent to which differences in alveolar pressures(PA) would develop between anemphysematous right lower lobe (RLL) and normal left lower lobe (LLL)during forced vital capacity (FVC) deflation. RLL and LLLPA(PA_RLL and PA_LLL,respectively) were measured by the alveolar capsule technique. Duringforced deflation, PA and lobarflows were determined between 95 and 20% FVC. A choke point common toboth lower lobes was observed at >40% FVC. The results showed thatdeflation compliance (C) was altered for the RLL such that <90%lobar vital capacity, C_RLL > C_LLL, whereas >90% lobar vitalcapacity, C_RLL < C_LLL. At 95 and 90% FVC, theinitial RLL PA decline wasgreater than that for the LLL (P < 0.05). However, large differences inPA were prevented because of theeffect of interdependence of regional expiratory flow (IREF). IREFcaused a relative decrease in RLL flows and increase in LLL flows thatlimited PA differences. Between 80 and 50% FVC, as C_RLL becamegreater than C_LLL, and because ofthe initial effect of IREF,PA_RLL was~PA_LLL.At 40% FVC, without IREF, lobar differences inPA widened. These findings indicate that IREF may affect the dynamics of flow limitation inregional lung disease.

     

The assessment of DNA from marine organisms via a modified salting-out protocol

We developed a rapid, practical and non-toxic salting-out method for the extraction of DNA from marine organisms, and tested it on two representative species of Porifera and Cnidaria, both living in association with symbiotic zooxanthellae. We tested the efficiency of the protocol by comparing the output of the method for fresh tissue, frozen tissue and tissue stored in ethanol. It proved to be effective for extracting DNA in the case of the methods of preservation considered here, and for obtaining quantities of DNA comparable to those obtained via the traditional approach. The DNA from both species was of good quality. The DNA obtained was amplified by PCR using specific primers for the large ribosomal subunit, allowing the identification of the presence of both the host and symbiont genomes.     

One preventable cause of spontaneous deflation of inflatable mammary prostheses.

Spontaneous "overnight" deflation of inflatable prostheses is rather uncommon, but we have had a 5.7% incidence of it in a 24-month period in which we used implants with a suturable tab and fastened them to the subjacent fascia. At exploration we found these tabs had torn the bag of the implant, usually at the vulcanized seam. We now believe that fixing a suture tab on a breast implant to underlying fascia may cause undue stresses upon the implant at that point and result in an otherwise avoidable deflation.     

Spleen and cardiovascular function during short apneas in divers.

We investigated the spleen volume changes as related to the cardiovascular responses during short-duration apneas at rest. We used dynamic ultrasound splenic imaging and noninvasive photoplethysmographic cardiovascular measurements before, during, and after 15-20 s apneas in seven trained divers. The role of baroreflex was studied by intravenous bolus of vasodilating drug trinitrosan during tidal breathing. The role of lung volume was studied by comparing the apneas at near-maximal lung volume with apneas after inhaling tidal volume, with and without cold forehead stimulation. In apneas at near maximal lung volume, a 20% reduction in splenic volume (P = 0.03) was observed as early as 3 s after the onset of breath holding. Around that time the heart rate increased, the mean arterial pressure abruptly decreased from 89.6 to 66.7 mmHg (P = 0.02), and cardiac output decreased, on account of reduction in stroke volume. Intravenous application of trinitrosan resulted in approximately 6-mmHg decrement in mean arterial pressure, while the splenic volume decreased for approximately 13%. In apneas at low lung volume, the early splenic contraction was also observed, 10% without and 12% with cold forehead stimulation, although the mean arterial pressure did not change or even increased, respectively. In conclusion, the spleen contraction is present at the beginning of apnea, accentuated by cold forehead stimulation. At large, but not small, lung volume, this initial contraction is probably facilitated by downloaded baroreflex in conditions of decreased blood pressure and cardiac output.     

The assessment of pulmonary vein potentials using the new achieve advance during cryoballoon ablation of atrial fibrillation

BackgroundThe new version of inner lumen mapping catheter (Achieve Advance?; Medtronic, Minnesota, USA) includes a new solid core which provides improved rotational response, as compared to the current Achieve Mapping Catheter. In the present study, we sought to analyze the rate of visualisation of real-time recordings using this new device comparing it with a large cohort of patients having undergone second generation cryoballoon (CB) ablation using the previous Achieve mapping catheter.MethodsAll patients having undergone CB ablation using the Achieve Advance and the last 150 consecutive patients having undergone CB ablation using the previous Achieve were analysed. Exclusion criteria were presence of an intracavitary thrombus, uncontrolled heart failure, moderate or severe valvular disease and contraindications to general anesthesia.ResultsA total of 200 consecutive patients (60.1 ± 9.5 years, 75% males) were evaluated (50 Achieve Advance and 150 old Achieve). Real-time recordings were significantly more prevalent in the “new Achieve Advance” population compared with the “old Achieve” group (73.5% vs 56.8%; p = 0.0001). Real-time recordings could be more frequently visualized in the “Achieve Advance” group in all veins except RIPV (LSPV: 86% vs 71.3%, p = 0.04; LIPV: 84% vs 62.7%, p = 0.005; RSPV: 78% vs 52%, p < 0.0001; RIPV: 46% vs 41.3%, p = 0.3).ConclusionsThe rate of visualisation of real-time recordings is significantly higher using the new Achieve Advance if compared to the previous Achieve mapping catheter in the setting of CB ablation. Real-time recordings can be visualized in approximately 73.5% of veins with this new device.     

A protocol for rapid isolation of flanking regions from short known sequences

We present a method for rapid isolation of flanking regions from amplified fragment length polymorphism (AFLP) fragments based on thermal asymmetric interlaced (TAIL)-PCR, in which one sequence-specific primer and one degenerate primer derived from an conserved motif found in homologies of the known sequence were used. The final result showed this to be a simple and efficient strategy, especially for short known sequences containing coding regions. Moreover this protocol was especially useful for species with little available genome information such as Hongkong Kumquat (Fortunella hindsii), since most of their genes have known homologies in other species such asArabidopsis and rice.